Telemetry User Manual[03-FEB-24] Our telemetry sensors and stimulators are battery-powered devices designed for use with small animals or fish. All of them use the same radio-frequency (RF) messaging system for transmitting information, so we can receive signals from all of them with the same telemetry receivers. Animals can be housed together or separately. A single telemetry receiver can record signals from dozens of sensors.
Our Subcutaneous Transmitters (SCT) are small, battery-powered devices designed for implantation beneath the skin of an animal. We turn them on and off with a magnet. Each SCT measures one or more biopotentials, and perhaps temperature as well. It samples biopotentials with sixteen-bit precision and transmits the samples by radio waves. Their batteries cannot be replaced. Our Head-Mounting Transmitters (HMT) mount on the exterior of an animal. They have no on-off switch, but we can remove and replace their batteries.
Our Implantable Inertial Sensors (IIS) transmit acceleration and rotation and turn on and off with a magnet. These devices are designed for implantation in animals or attachment to fish. Our Implantable Stimulator-Transponders (ISTs) hibernate when placed next to a magnet, but otherwise are activated and controlled by radio waves. The RF messaging system we use for transmitting commands to stimulators is distinct from the one our sensors use to transmit information, but it does use of the same antennas.
A Telemetry System consists of sensors, Faraday enclosures, receivers, and acquisition computer. The computer and receiver communicate with one another by TCPIP. We have two types of receiver. A coaxial antenna receiver gathers telemetry signals with antennas on the end of coaxial cables. A coil array receiver gathers telemetry signals with antennas fixed in an array on a platform. The Animal Location Tracker (A3038, or ALT) is a coil array receiver that we connect to Power over Ethernet (PoE). The Telemetry Control Box (TCB) is a coaxial antenna receiver. The older Octal Data Receiver (ODR), now discontinued, is a coaxial antenna receiver that requires a LWDAQ Driver to provide power and Ethernet.
Our Faraday enclosures isolate the telemetry system from ambient RF interference. Before investing in one of our telemetry systems, send us your animal laboratory's latitude and longitude so we can check for nearby mobile phone base stations. We will consult a global database to see if there are any base stations near your building. We recommend you do not attempt to operate our telemetry system within 50 m of a 6-kW base station or within 20 m of a 1-kW base station.
The TCB provides sixteen antenna inputs to which we can connect up to sixteen Loop Antennas (A3015C) for telemetry reception. We can place four such antennas in each of four bench-top Faraday enclosures. Or we can place sixteen antennas in a single Faraday canopy containing shelving units holding animal cages. In each case, we can record from dozens of animals. The ALT requires no external antennas. It will record from whatever transmitters share its Faraday enclosure, or we can configure the ALT to record from only certain transmitters. The ALT provides, in addition to telemetry signals, a measurement of the position of each transmitter on its array of detector coils.
Our SCT and HMTs will monitor electroencephalograph (EEG), local field potential (LFP), electrocardiogram (ECG) or electromyography (EMG) at frequencies down to direct current (DC) and up to 640 Hz. We describe the sources of EEG and LFP in The Sources of EEG. For recordings from live animals, see our Example Recordings page. We currently have three models of SCT in production, each of which is available in a large number of variants. The A3048 provides only one input channel Its amplifier provides gain of ×100 for frequencies up to 160 Hz. Our smallest SCT is the A3048P with volume 0.70 ml and mass 1.8 g. We mount the battery beside the A3048 circuit rather than on top of it, which results in the A3048 having greater surface area, while being both thinner and lighter. The A3049 provides two input channels. These may be configured to each use their own reference potential or to share a reference potential. The A4049 amplifiers provide gain ×100 at frequencies up to 640 Hz. The A3049 can be equipped with small batteries for use in mice, or large batteries for use in adult rats. The A3047 if a four-channel transmitter that incorporates its own temperature sensor. Each of the four inputs will operate with its own reference potential or a shared reference potential. The amplifiers provide gain ×100 up to 640 Hz, for which the sample rate is 2048 SPS. The A3047 is intended for use in adult rats.
The primary purpose of our telemetry system is to record biopotentials continuously for weeks at a time, provide intermittent stimuli for weeks at a time, and to analyze the recordings automatically. When recording EEG in epileptic animals, we want to count seizures in tens of thousands of hours of recording. The A3049A3 provides two input channels that share the same reference potential, allowing EEG to be monitored in two locations. Its volume is 1.2 ml, which fits easily into an adult mouse. It samples its Red and Yellow inputs at 512 SPS each, and its amplifiers provide bandwidth 0.3-160 Hz. It runs for 14 days. The Red and Yellow inputs share the same common terminal, the Blue input, so there are three leads emerging from the device, plus the antenna. The A3049B3 is the same size as the A3049A, but provides only the Red input and runs for 24 days. The A3049D3 is electrically identical to the A3049A3, with a larger battery. It's volume is 3.0 ml and it runs for 76 days. The A3049E is also 3.0 ml, but provides only the Red input and runs for 132 days. The A3047A1A provides three inputs, each with its own reference potential, and is designed to record EEG (electroencephalogram), EMG (electromyogram), and EGG (electrogastrogram) all at the same time. It runs for 76 days.
Reception of our telemetry signals is always limited by RF interference. Our faraday enclosures reduce ambient interference by a factor of one thousand. Where a single antenna receives the signal from an implanted transmitter 95% of the time, two independent antennas pick up the signal 99.5% of the time. Our Octal Data Receiver (A3027) provides eight independent antenna inputs. Each input has its own amplifier, demodulator, and decoder. By distributing these antennas between our animal cages, the probability that at least one of them will receive any particular message approaches 100%. With eight antennas distributed through an IVC rack, for example, we obtained 100% reception everywhere within the rack, despite the presence of a 900-MHz mobile phone base station 80 m from the recording room. One Octal Data Receiver can record from dozens of animals in one such IVC rack. Our FE3A enclosure stands on its own, and is 90 cm wide, 60 cm high, and 65 cm deep. With a shelf half-way up, it holds six animal cages. With four antennas inside, it will provide robust recording from forty co-habiting animals.
Faraday enclosures and multiple antennas cannot overcome the most severe interference a crowded city can produce. In our laboratory, a 1-W interference source at range 1 m will cause 50% loss of signal from a moving transmitter in a faraday enclosure equipped with two independent antennas. By application of the inverse square law, we expect a 3000-W 900-MHz GSM base station 55 m away to be capable of similar disruption. The power emitted by a mobile phone base station spreads out like the beam of a flash-light. If you are nearby but below the station, you can receive less power than if you are farther away but at the same altitude, and if the station is on a neighboring building, you will receive more power than if it is mounted on your own building. If your recording room is less than 100 m from a base station, and directly in the path of its transmit beam, we will advise you on how to make sure your telemetry system will provide robust reception.
You will find the prices of subcutaneous transmitter system components in our price list. Our Event Detection page gives a history of our automatic event detection software. In the body of this report, we describe the performance and operation of our Subcutaneous Transmitter hardware. In places, the description is detailed and technical. Consult our terminology page for definitions of terms like RF, IF, LO, mixer, attenuator, dBm, and dB.
[10-FEB-24] Our transmitters provide one or more telemetry signals, depending upon how they are configured. We have two classes of telemetry receiver : coaxial antenna and coil array. The coaxial antenna systems use antennas on the end of coaxial cables. We distribute the antennas among our Faraday enclosures, placing them between and adjacent to our animal cages. The coil array systems provide a platform upon which we can place one or two animal cages. Beneath the platform is an array of small antennas.
| Assembly | Name | Type | Description |
|---|---|---|---|
| A3018 | Data Receiver | Coaxial Antenna | Single antenna amplifiers, requires LWDAQ Driver, discontinued 2016. |
| A3027 | Octal Data Receiver (ODR) | Coaxial Antenna | Eight antenna amplifiers, requires LWDAQ Driver, discontinued 2022. |
| A3071 | LWDAQ Driver | TCPIP Interface | Required by A3018 and A3027 receivers, discontinued 2022. |
| A3038 | Animal Location Tracker (ALT) | Coil Array | 48 cm × 24 cm array, location tracking, PoE, active. |
| TCB-A16 | Telemetry Control Box (TCB) | Coaxial Antenna | 16 antennas, location monitoring, PoE, active. |
Our coaxial antenna systems will each record from a hundred transmitters, making them cost-effective. Our coil arrays provide location tracking with sufficient accuracy to allow robust activity measurement and reliable disambiguation of animals in video recordings. Our new Telemetry Control Box (TCB) provides approximate location tracking for multi-room habitats, as well as location monitoring for animals in cages.
Our well-established coaxial antenna system consist of an Octal Data Receiver (A3027, ODR) and a LWDAQ Driver. The ODR comes in a 30 cm × 22 cm × 11 cm metal box. The box connects to the LWDAQ Driver with a shielded network cable. It receives power through the same cable. Eight BNC sockets on the back of the box provide connections for a coaxial cables to the antennas. There are thirty indicator lights on the front of the box. There is one white light for each transmitter channel, one white light for each antenna input, and power, reset, upload, and empty lights. The Octal Data Receiver comes with eight Dampled Loop Antenna (A3015C). These attach to the antenna inputs with coaxial cables that pass through a Faraday enclosure wall with the help of a BNC feedthrough. Each antenna can stand on its own, or we can take the mounting brackets off and lay it on a table or wedge it between the cages in an IVC rack.
The Telemetry Control Box (TCB-A16) eliminates the separate LWDAQ Driver. It requires only one Power over Ethernet (PoE) connection for power and communication combined. It provides sixteen antenna inputs, each with its own independent telemetry receiver, and can be configured to monitor the telemetry signal power received by each of its antennas, so as to permit us to estimate which of the sixteen antennas is nearest to the transmitter, and to monitor the activity of animals in cages. The planned Telemetry Control Box (TCB-B16) adds sixteen command transmitters to control devices like the Imlantable Stimulator-Transponder (A3041).
Our Animal Location Tracker (A3038, ALT) connects to PoE for power and communication. A single ALT permits recording from all animals moving over its coil array. The A3038A platform is 51 cm × 27 cm, so we can place one large rat cage or two small mouse cages upon a single platform.
| Assembly Number and Manual Link | Assembly Name | Status |
|---|---|---|
| A3028 | Subcutaneous Transmitter | Discontinued |
| A3048 | Subcutaneous Transmitter, One Input | Active |
| A3049 | Subcutaneous Transmitter, Two Input | Active |
| A3047 | Subcutaneous Transmitter, Four Input with Temperature | Active |
| A3015C | Loop Antenna | Active |
| A3039 | Coaxial Feedthroughs and Combiners | Active |
| FE3A | Bench-Top Faraday Enclosure | Discontinued |
| FE3B | Bench-Top Faraday Enclosure | Active |
| FE5A | Canopy Faraday Enclosure | Active |
| A3038 | Animal Location Tracker | Active |
| TCB-16A | Telemetry Control Box | Active |
| TCB-16B | Telemetry Control Box | Planned |
| A3027 | Octal Data Receiver | Discontinued |
| A2071E | LWDAQ Driver | Discontinued |
Faraday enclosures can be bench-top with hinged doors, or they can be canopies supported by an aluminum frame around a free-standing rack of shelves. The bench-top enclosures have back walls made of aluminum sheet where we place coaxial and Ethernet feedthroughs to carray coaxial and Ethernet connections. The canopies have floors made of aluminum sheet to which we attach coaxial and Ethernet feedthroughs.
[17-OCT-23] Before investing in one of our telemetry systems, we suggest you send us your animal laboratory's latitude and longitude so we can consult the global mobile phone base station database for transmitters near your facility. Mobile phone networks that use the 902-928 MHz band are less common now than they were ten years ago, but they still exist. If you have one on the building opposite you, directing power right through your windows, your telemetry system will not perform well.
We recommend you set up your telemetry system several weeks before you plan to perform your first animal experiments. During these weeks, you can install our software on your computer and learn how to acquire and analyze telemetry signals. We supply one or two sample transmitters with your recording system, so use these to test reception and recording. Place one or two transmitters within a Faraday enclosure and record their signals continuously for a week to make sure that your computer, local area network, and hardware is functioning correctly. Recording might be interrupted every few hours, and you will be glad to figure out what causes these interruptions before you have animals with transmitters implanted. Reception from the transmitters may cease all-together every few hours, and you will be glad to figure out what is interfering with reception long before you have begun an experiment with live animals.
List of parts for coaxial antenna system with Telemetry Control Box:
List of parts for coil array recording system with Animal Location Tracker:
List of parts for coaxial antenna recording system with Octal Data Receiver:
Assuming you have the above parts in hand, follow these steps. They will take you through the set-up of the hardware and software, so that by the end you will be able to record and analyze transmitter signals.
When our telemetry system consists of several receivers, we will need one Neurorecorder for each. If we want to process the signals as they are recorded, we will need at least one Neuroplayer per receiver. We may even need one Neuroplayer per animal. For example, we may have two animal location trackers (ALTs) with four animals over each tracker, and we want to export the signals from each animal to separate EDF files. We need two Neurorecorders and eight Neuroplayers. The Startup Manager allows us to define our telemetry recording and processing with a script and start all processes by running the script. After an interruption, we can re-start the system with a few mouse clicks.
[30-NOV-21] We have a library of example telemetry recordings available on our example recordings page. You will also find two short example recordings in the LWDAQ/Images folder: they are the two files with extension "ndf".
For an totorial on browsing recordings with the Neuroplayer, see our Neuroplayer Introduction video.
[09-FEB-24] Our telemetry system supports four types of activity monitoring. None of the measurements are perfect, but all are useful. The four types are: location monitoring, location tracking, acceleration recording, and acceleration with rotation recording. The first two are provided by the telemetry receiver. The second two are provided by a dedicated implant that holds an accelerometer and gyroscope: the Implantable Inertial Sensor (IIS).
The Telemetry Control Box (TCB) measures the radio-frequency power received by each its antennas whenever it receives and decodes a telemetry message. It provides to us the number of the antenna that received the greatest power and a logarithmic measurement of that power. We call these the top antenna and the top power. In a telemetry system in which the antennas are separated by at least thirty centimeters, the top antenna is almost certainly the one closest to the transmitter. Thus the TCB allows us to determine the location of an animal in a maze or some other environment with multiple chambers. We call this location monitoring.
The Animal Location Tracker (ALT) measures the radio-frequency power received by each of its detector coils whenever it receives and decodes a telemetry message. It provides us with a logarithmic measurement of this power at each of its coils. By taking a weighted centroid of the receive power, the ALT provides us with a location. This location is not an accurate measurement of the location of an animal, but its movement is well-correlated with the movement of the animal, and its value is well-correlated with position. We can use the centroid to obtain a measurement of total distance traveled by each animal in a cage. We can measure the time pairs of animals spend close together. We can use the movement of the centroid to determine which animal is which in continuous video recordings. We call this location tracking.
Both location monitoring and location tracking come at no cost in operating life of the transmitter. We can perform both measurments with any telemetry device that transmits telemetry messages. If we want a higher-resolution measurement of the acceleration of an animal, we must implant an accelerometer. We will obtain 64 SPS of acceleration measurements in three orthogonal directions. We call this measurement acceleration recording. The accelerometer-only versions of the IIS provide acceleration recording. If we want to measure acceleration and rotation, the accelerometer with gyroscope versions of the IIS provide acceleration with gyroscope recording. The gyroscope is power-hungry, so the IIS with gyroscope supports intermittent measurements over a long period, but not continuous measurements.
[04-MAR-16] The Subcutaneous Transmitter (A3019) provides one analog input channel by means of two flexible, insulated stainless steel flexible wires. In the A3019A, this input is low-pass filtered by a three-pole active filter with roll-off of at 160 Hz and high-pass filtered by a single-pole filter with roll-off at 0.2 Hz. Its dynamic range is 20 mV. The A3019A can detect a 5-μV sinusoid of any frequency between 5 Hz and 160 Hz, and measure its amplitude with 1-μV accuracy. The A3049 is a two-channel transmitter with similar performance, depending upon how it is configured.
All transmitter models digitized at with sixteen-bit precision. The A3019A and A3028A digitize each input channel at 512 SPS. Each sample is transmitted in its own, isolated, 7-μs radio-frequency message. Transmission takes place through a dedicated loop antenna. The A3019A and A3028A transmit in the 902-928 MHz ISM band. It uses two frequencies to represent two logic levels. The messages propagate through space as radio waves. See How Antennas Work in our Technical Proposal for an explanation. The power transmitted by the antenna during the 7-μs burst is roughly 300 μW (−5 dBm).
The transmitter's analog circuits operate continuously to amplify and filter its analog signals. But the logic and RF circuits are inactive almost all the time. In the case of our A3019A, the logic and RF circuits wake up for 7 μs every 1953 μs, which is 0.36% of the time. During this 7-μs burst of activity, the transmitter converts its single analog input into a sixteen-bit number, and transmits this number, together with some synchronizing bits, the transmitter's four-bit ID, and a checksum. We call this transmission a message. The A3019A's message rate is 512 per second. Within each message, the transmitter sends bits at the bit rate. The A3019 and A3028 bit rate is 5 MBPS, or 200 ns per bit. For more details of the message encoding, see below.
The radio-frequency (RF) messages are received by our Loop Antenna (A3015). The messages are joined in the antenna by RF signals from other sources of similar frequency. The messages and the interference propagate along a coaxial cable to the Data Recorder (A3018) or Octal Data Receiver (A3027). The signal power arriving at the antenna must be at least four times (12 dB) greater than the interference power from other radio-frequency sources, and at least 25 pW (−76 dBm) if there is no interference. Interference power of 10 nW (−50 dBm) in the 902-928 MHz band is common in urban areas, at which level, the receive antenna must receive at least 40 nW, which we cannot rely upon when the transmitter is implanted in a moving animal, even if the receive antenna is no more than 30 cm from the animal. In order for reception to be reliable in the presence of 10 nW of interference, we must place the transmit and receive antennas inside a Faraday enclosure to reduce the interference power below 1 nW. Faraday enclosures such as our FE2F provide at least a factor of ×1000 (30 dB) isolation from external interference, and so make it possible to obtain reliable recordings with one antenna from up to fourteen cohabiting animals. Larger, less restrictive Faraday enclosures offer isolation of 10 dBm, and with the help of multiple, independent antennas connected to the same receiver, we obtain reliable reception despite prominent interference.
The radio signal enters the metal enclosure of the receiver and connects to the RF input of a demodulating receiver (such as the A3017). The RF messages and interference are amplified by 20 dB and passed through a bandpass filter that rejects signals outside the 902−928 MHz. What remains are the messages and ISM-band interference. These are amplified by another 20 dB and enter a mixer, which downshifts the 902-928 MHz RF signal to a 38-64 MHz intermediate-frequency (IF) signal.
The IF signal passes through three limiting amplifiers. Each provide 22 dB of gain, and limit their output. When we disconnect the antenna, the signal on IFL is random, but not quite saturated. When we connect the antenna, the RF interference in our laboratory causes IFL to saturate almost all the time. Because IFL is saturating, its amplitude is constant. Only its frequency changes. The fixed-amplitude IFL passes into a tuned circuit called the discriminator. The discriminator attenuates the lower IF frequencies, so that its output amplitude depends upon the frequency of IFL.
The discriminator output passes into a full-wave demodulator. The demodulator turns the AC signal into a signal proportional to the AC amplitude. The demodulator output is D in the S3017_2 schematic. The alternating radio frequencies of the transmitter messages appear in D as a square wave.
The D signal is mildly band-pass filtered by two RC networks to form the signal S in the S3007_1 schematic. The transmitter messages appear in S as a square wave centered about the 0-V potential.
A comparator transforms S into a logic HI when S is above 0 V, and logic LO when it is below 0 V. The comparator output is called C. The logic levels of the original transmitter message now re-appear as logic levels on C. Outside the messages, C is a random or pseudo-random sequence of logic levels. It is random when the dominant source of RF power at the antenna input is electronic noise, and pseudo-random when the dominant source is interference in the ISM band.
The C logic signal passes into the receivers's logic chip, where it is synchronized with a 40-MHz message clock. The synchronized version of C is SC. It changes only on the rising edges of the message clock. Pulses on C that take place between these rising edges do not appear in SC. The receiver monitors the stream of bits on SC, looking for messages embedded within the stream. These messages could arrive at any time. Messages can also appear in the SC bit sequence by chance, as a result of thermal noise or interference. We call these bad messages.
The receiver stores any messages it detects in its 512 KByte first-in first-out buffer as thirty-two bit records, as we defined by our telemetry message encoding. The last eight bits are a timestamp, which counts cycles of the receiver's 32.768 kHz clock.
The receiver stores clock messages 128 times per second in among the telemetry messages. The clock messages take the form of a transmitter message from a transmitter with ID zero. The sixteen-bit data of a clock message is a sixteen-bit counter that increments by one with every clock message stored. The final eight bits are a timestamp, which is always zero, because the clock messages are stored whenever the receiver's 32.768 kHz cycle counter wraps around to zero. You can think of the clock messages as being the product of a virtual transmitter with ID zero, transmitting 128 messages per second, and whose data is a counter that increments from one message to the next.
The receiver connects to the LWDAQ. The LWDAQ in turn connects to an Ethernet. Your data acquisition computer runs the LWDAQ software and communicates with the LWDAQ by TCPIP.
The LWDAQ software on your data acquisition computer uses its Receiver Instrument to download blocks of messages from the receiver's first-in first-out buffer. It plots these messages on the screen, including the clock messages. The Recorder provides you with blocks of data that cover a time interval you specify. As it acquires data, the receiver makes no effort to eliminate bad messages or insert substitute messages. Its data blocks are therefore of variable memory-size, but fixed time-duration.
The Neurorecorder Tool calls the Receiver Instrument to supply fixed time-duration data blocks. The Neurorecorder records all data downloaded from the receiver and stores it to a file. The file contains the raw values downloaded, with no processing or alterations made, and also provides a substantial metadata field for text data describing the recording. The file is arranged in our general-purpose NDF file format.
Meanwhile, the Neuroplayer will read, process, and display recordings from any NDF file, including the one that is receiving the freshly-downloaded data. We tell the Neuroplayer which channel numbers we want to reconstruct and display. Each channel corresponds to a transmitter recording. A two-channel transmitter will have two consecutive channel numbers for its two signals. Our subcutaneous transmitters have channel numbers between 1 and 222, excepting any number with remainder zero or fifteen after division by sixteen. Channel numbers with remainder zero are reserved for metadata. Channel number zero itself contains the receiver's clock messages. Channel numbers with remainder fifteen are reserved for auxiliary messages, which are solitary messages such as command acknowledgements and battery measurements. The reconstruction of each channel from the raw data involves removing bad messages and inserting substitute messages where necessary. The Neuroplayer displays the reconstructed channels and their discrete Fourier transforms. When playing back an archive, the Neuroplayer will perform processing on the signals, so as to calculate metrics for event classification, export the signal to another recording format, or determine the total power in various bands of the signal spectra.
The bit rate is the rate at which the transmitter sends bits during one of its message transmissions. The A3019A, for example, transmits a total of 40 bit-values in 8 μs. The bit rate is 5 MBPS (megabits per second). We describe the function of these bits in the Message Encoding section here.
With the exception of a few temporary, slower versions we made for the following tests, all Stage Four and Stage Three circuits run at 5 MBPS. Here we compare the performance of the Stage Four system at 5 MBPS and 2.5 MBPS and show that the gain in operating range we make by dropping the bit rate is slight, while the loss in operating life is significant.
We set up a Demodulating Receiver (A3017) with a SAW Oscillator (A3016SO) and Data Recorder (A3007C). We connected a Dipole Antenna (A3015B) on a 240-cm (96") coaxial cable (RG58C/U). We recorded messages from a Subcutaneous Transmitter (A3013). We configure the transmitter and receiver for message transmission at 5 MBPS (200 ns per bit), then for 2.5 MBPS (400 ns per bit). The switch between bit rates requires the following changes.
We measure a variety of performance parameters for each bit rate. We present our measurements in the following table.
| 2.5 MBPS | Parameter | 5 MBPS |
|---|---|---|
| 918±4 MHz | Frequency Modulation | 918±4 MHz |
| 13 MHz | Bandwidth (90% Power) | 18 MHz |
| 12 μs | Message Duration | 8 μs |
| 108 μA | Active Current | 75 μA |
| 18 μA | Inactive Current | 18 μA |
| 15 m | Maximum Range (favorable orientation) | 10 m |
| <0.1/s | Bad Message Rate (Antenna Disconnected) | <0.1/s |
| <0.1/s | Bad Message Rate (Transmitter Inactive) | <0.1/s |
| <0.2% | Blocked Message Rate (Transmitter Adjacent to Antenna) | <0.2% |
| ≈3% | Missing Message Rate (Transmitter Moving Randomly 1 m from Antenna) | ≈2% |
| ≈15% | Missing Message Rate (Transmitter Moving Randomly 1 m from Antenna and Enclosed Between Two Hands) | ≈25% |
The 5 MBPS and 2.5 MBPS transmissions perform best with a modulation depth of ±4 MHz. The 90% power bandwidth of the 5 MBPS messages is 18 MHz, which leaves us with at least 8 MHz of extra room in the 902-928 MHz band. The power bandwidth of the 2.5 MBPS messages is 13 MHz. In theory, the narrower the bandwidth, the less likely a hole in the reception caused by interference (such as the this) will lie within the transmission bandwidth, and therefore the less likely that the transmission will be corrupted by interference.
The biggest benefit of the lower bit rate is a 50% increase in operating range. The biggest cost of decreasing the bit rate are the increase in active current. The current rises from 75 μA to 108 μA. The operating life drops from nine weeks to six weeks. Another cost of decreasing the bit rate is the increased transmission time, which means a greater probability of collisions between multiple transmitters sharing the same receiver. The probability of collision between any two transmitters on any given message transmission is 0.7% at 5 MBPS and 1.4 % at 2.5 MBPS. With ten transmitters, each would lose 7% of its samples at 5 MBPS and 14% at 2.5 MBPS. Our system can handle a 30% loss of messages. At 2.5 MBPS we would use up most of this missing message budget in the normal operation of ten transmitters.
We decided to stay with a 5 MBPS bit rate. Our collaborators at ION stated that battery life and bandwidth were more important than operating range, because in either case the operating range appears to be greater than the size of a standard rat cage.
[27-SEP-23] The subcutaneous transmitters encode ones and zeros in a frequency-modulated radio signal. The signal consists of two frequencies separated by roughly 8 MHz, and both within the 902-928 MHz band. The transmitter messages are a sequence of ones and zeros sent with Manchester Encoding. We first described the telemetry message structure here. Each message begins with eleven bits of value one (1). These bits serve two purposes. They synchronize the receiver clock with the message clock, and their exact period serves to distinguish the start of a transmitter message from random noise. The message body begins with a start bit of value zero (0). After the start bit comes the lower four bits of the transmitter's eight-bit channel number. The most significant bit comes first. The sixteen-bit sample comes next, most significant bit first. After the sample comes a four-bit completion code. The completion code is a function of the channel number, and allows us to deduce the upper four bits of the channel number from the lower four bits.
Where C is the completion code, N is the lower four bits of the channel number, and S is the upper four bits of the channel number, also called the set number. It would have been simpler to replace the completion code with the set number in the message encoding, but this completion code definition allows us to retain backward compatibility with our original four-bit channel number protocol. The four-bit system is equivalent to a system that uses only set zero. The following table gives examples of set numbers, channel numbers, and completion codes.
We reserve channel number 0 for clock messages, and call it the clock channel. Clock messages are generated by the receiver only. Clock messages are stored among the transmitter data messages. We reserve all channel numbers for which the lower four bits are 0 for internal use. We reserve all channel numbers for which the lower four bits are 15 for auxiliary data and we call these the auxiliary channels. Each set has its own auxiliary channel. We reserve channel numbers 225-254 for later use.
We note that the message contains no checksum to help detect corruption of the message contents. The primary means by which the receiver picks out genuine messages from interference and noise is through close examination of the timing of the signal. The encoding of the message produces a frequency transition every 195-215 ns. If a transition fails to occur, the receiver will abort decoding and look for another sequence of sychronizing bits. In our experience, each bit that satisfies the timing constraint makes it three times less likely that the message will turn out to be a bad message arising from interference or noise.
In the Receiver Instrument and the Neuroplayer Tool, we refer to signals by their channel numbers. The Receiver Instrument allows us to configure receivers to select only one set, which we specify with a set number. By default, the Receiver Instrument attempts to configure its receiver to record from all sets.
Note: We introduced channel numbers 16-222 in January 2017. A firmware upgrade will permit any receiver to record from the higher channel numbers. An Octal Data Receiver (A3027E) with firmware version ≥11 can be configured to receive from any single set zero (0) through thirteen (13), or from all sets zero through thirteen (0-13) simultaneously. We configure the receiver with the daq_set_num parameter in the Receiver Instrument. When this parameter is 0-13, the receiver accepts messages only from the corresponding set. When this parameter is "*", the receiver accepts messages from all sets.
Receivers store transmitter messages as four-byte records. The first byte is the transmitter channel number. The next two bytes are the sixteen-bit data and the last byte is a timestamp. The timestamp counts periods of the receiver's 32.768 kHz clock. The timestamp returns to zero 32768 ÷ 256 = 128 times per second. Whenever the timestamp returns to zero, the receiver inserts a clock message into the data. The clock message has zero for its channel number, this being the clock channel. Its sixteen data bits are a counter that increments by one from one clock message to the next. Its final byte contains the receiver firmware version number.
The auxiliary channels allow devices such as the Implantable Stimulator-Transpponder (IST, A3041) to transmit acknowledgments, battery measurements, and other metadata through our telemetry receivers. When the device transmits an auxiliary message, it does so using the nearest auxiliary channel above its primary channel number. The lower four bits of an auxiliary channel number are all ones, representing the number fifteen. The Receiver Instrument identifies all such messages as they arrive, and places them in a list. These four bits are followed by sixteen content bits, as in any of our telemetry messages. After the content come four completion-code bits. The leading ones and the completion code specify the auxiliary channel number. The sixteen content bits begin with a four-bit identifier. These identifier is intended to disambiguate the source of an auxiliary message. Following the four-bit identifier is a four-bit field address. The field address is intended to indicate what kind of information the source is trying to convey. The remaining eight bits of the auxiliary messagea are the data byte, which contains the information.
The Receiver Instrument supports auxiliary channels by diverting their contents into an auxiliary message list. Each element in the list is itself a list of four numbers: the channel number of the signal source that generated the auxiliary message, the field address, the data bits, and a timestamp. The timestamp is the value of the receiver's 24-bit clock, which counts up at 32.768 kHz, so that the precision of the clock is ±15 μs and the clock cycle is 512 s.
Example: Devices like ISTs have sixteen-bit identifiers. We make sure the lower four bits are never zero or fifteen. When an IST transmits an auxiliary message, it uses the auxiliary channel number we obtain by taking the lower byte of its identifier and setting the lower nibble to all ones. The IST with iedntifier 0x0B56 uses auxiliary channel 0x5F. Within the auxiliary message, the IST uses 0x6 for the identifier. We can deduce the lower eight bits of the IST identifier from the auxiliary message alone. The field address and data byte convery an acknowledgement, battery measurement, a state transition, or some other such information. Immediately following the first auxiliary message, the IST transmits another auxiliary message that acts as a confirmation. The confirmation message uses the same auxiliary channel and provides the same identifier, but its data byte is the upper byte of the IST's sixteen-bit identifier. When we receive two auxiliary messages from the same auxiliary channel within one clock tick of one another, the confirmation coming second, we are confident that we have received a genuine report from an IST, and we know that IST's identifier.
[12-JUL-23] Our telemetry system records its telemetry signals to disk in files that confrom to our Neuroscience Data Format (NDF). An NDF file begins with the characters " ndf" followed by a four-byte metadata string address, a four-byte data address, and a four-byte metadata string length. The byte ordering is big-endian (most significant byte first). The telemetry data is in the data section, with one record saved per unique message received from the telemetry system. We describe the message format in detail elsewhere, but we will summarize the format here. Each message consists of a core and a payload. The core is four bytes long. The first byte is the telemetry channel identifier. The next two bytes are the message data value, which almost always be a sixteen-bit sample value with its most significant byte first. The fourth byte of the core is a timestamp. The payload consists of further bytes of information obtained from the telemetry receiver. Some receivers produce no payload. Others produce payloads up to sixteen bytes long.
| Receiver | Payload (Bytes) |
Description |
|---|---|---|
| A3018 | 0 | Data Receiver, no payload |
| A3027 | 0 | Octal Data Receiver, no payload |
| A3038 | 16 | Animal Location Tracker, sixteen detector coil powers |
| A3042 | 2 | Telemetry Control Box, top power and top antenna |
The program that does the recording is the Neurorecorder, which is a tool build into our LWDAQ software. The Neurorecorder writes "payload" field to the NDF metadata string, in which it notes the length of the message payload in the NDF file's messages. The Neurorecorder knows the payload length because it queries the receiver to determine the receiver version, and from this version number the Neurorecorder deduces the message payload length. Here is an example metadata string from an NDF recorded from an Animal Location Tracker (ALT).
When we play an NDF recording in the Neuroplayer, the Neuroplayer reads the payload length from the metadata and acts accordingly. The payload provided by an ALT contains the power of the message at each of the ALTs location and auxilliary detectors. The power bytes, combined with the spatial distribution of the ALT's detector coils, allows us to deduce the approximate location, and measure the activity of animals. The "alt" field in the metadata gives the locations of the detector coils in a three-dimensional coordinate system. The payload provided by the TCB gives us the maximum power with which the message was received by any of the TCB's detector coils, and the identifier of that detector coil. The "alt" field for a TCB recording would likewise provide us with the locations of the detector coils, and the top antenna will give us an idea of where the animal is in a large space, or a multi-room habitat, in which we have one or two antennas in each room.
[12-JUL-23] We define reception robustness as the fraction of the time during which we receive 80% or more transmitted messages while the transmitter rotates and moves randomly at a particular range. Our sample rate and filtering is designed to tolerate the lost of 20% of the samples without loss of signal quality. We define robust reception as reception with robustness 95% or higher. In other words: signal quality is degraded by loss of samples for less than 5% of the time. A transmitter's operating range is the maximum range from the receiving antenna at which reception is robust. We use Faraday enclosures and multiple, independent antennas to guarantee robust reception throughout the interior of the enclosure in all laboratory environments, regardless of ambient interference.
The receiver can fail to identify the message in its incoming bit stream (see Signal Path), even though they are received clearly by the Demodulating Receiver. The receiver might be occupied with something that looks like a message, and when the real message arrives, all it knows is that the previous message was invalid. The receiver goes back to looking for a new message, but is too late to identify the real message. We measure the rate at which the receiver loses messages by putting a single transmitter next to the antenna and watching for missed messages. Our signal is strong and there are no other transmitters to collide with it. We assume that any missing messages have been lost in the receiver. It turns out that the number of synchronizing bits that the receiver requires at the beginning of a message is what dictates the missing messages rate.
| Number of Synchronizing Bits | Missing Message Rate |
|---|---|
| 2 | 1.2% |
| 4 | 0.5% |
| 5 | <0.2% |
Only after it has received its required number of synchronizing bits does the receiver proceed to message recording, and it will lose an incoming message only if it is already recording a false message. By insisting upon five synchronizing bits, we ensure that the receiver loses less than one in a five hundred messages. As it turned out, we settled upon 11 synchronizing bits because this improved the performance of the system when the signal was weak.
We placed a single transmitter within a Faraday enclosure, on an upturned paper cup 10 cm from the antenna within a Faraday enclosure. We recorded continuously for an hour. Average reception was 99.7%. When we turn off the transmitter, the bad message rate is less than 0.1%. We conclude that roughly 0.3% of message are being blocked.
Each transmitter, when active, transmits messages in a continuous stream. The A3019A transmits 512 sixteen-bit data samples per second. A message that the receiver fails to receive is a missing message.
The Neuroplayer Tool handles missing messages by creating a substitute message with equal data sample value to the previous message received from the same transmitter. It does this only for the channels you are recording.
Despite the Neuroplayer's message substitution, missing messages degrade the effective bandwidth and quality of our received signals. But they do not cause catastrophic problems, nor do they confuse our data acquisition system, which can rely upon a guaranteed stream of messages from each transmitter it monitors.
The primary cause of missing messages is lack of signal strength at the antenna when compared to ambient interference power, as we discuss in Ambient Interference. The primary cause of loss of signal strength is cancellation of the RF signal by its own reflections arriving at the antenna, as we discuss in Operating Range. The primary source of ambient interference is cordless phones operating in the 902−928 MHz band, and mobile phones operating in the frequency bands immediately above and below the 902−928 MHz band. Even if such mobile phones transmit only 1% of their power outside their designated frequency bands, they will still combine to produce substantial interference in neighboring bands. Another source of interference is other subcutaneous transmitters, in which case we refer to the interference as a collision.
Mobile phone interference in the ION laboratory in London, which is on the eighth floor of a building, was so severe that reception from an implanted transmitter at range 50 cm would sometimes drop as low as 20%, and was rarely above 80%. With the help of one of our Faraday enclosures, reception jumped up to an average of 99%, with a minimum of 98% during any four-second period.
Noise and interference can generate messages on their own. We call these bad messages. Bad signals can raise false alarms, spoil the scale of self-adjusting displays, and wreak havoc with your Fourier Spectrums. We have made every effort to avoid them.
Bad messages have a valid transmitter ID number, but their data is invalid. The Receiver Instrument rejects them when they occur with ID numbers you have not asked it to record.
The bad message rate in our laboratory is less than one per ten minutes when there are no transmitters active. In London, the bad message rate was roughly one per second, sometimes coming in bursts of ten in a second. With transmitters active, we find that some messages get corrupted, as we describe below, and these message become bad messages.
We spend the rest of this section showing how bad message can, in theory, arise from noise and interference, and how our message encoding makes such bad messages very unlikely.
As we describe in Signal Path, the receiver monitors a logic signal called SC, looking for messages. These messages could arrive at any time. If the stream of bits is random, there is a chance it will generate a message by chance. If the stream of bits has some pattern to it because of radio interference, the chance of a message appearing in it at random might be much higher.
When we disconnect the receiver's antenna, and replace it with a 50-Ω terminator, we stop radio signals from reaching our receiver. All that remains at the input to our RF amplifier is white noise, whose power in our radio-frequency passband is around −87 dBm. The receiver amplifies this noise until it is large enough to generate an active pattern of zeros and ones on SC. This alternation is random, because the white noise is random. The output of the Demodulating Receiver (A3017) is an analog signal, S. This signal is sharply bandwidth-limited by the Demodulating Receiver's radio-frequency passband, and mildly high-pass filtered when it enters the receiver. The receiver uses a comparator to generate the logic level SC from S. When we examine SC on an oscilloscope with the antenna disconnected, the bandwidth of the bit stream is around 10 MHz, which we determine with the help of various low-pass filters.
Our messages represent individual bits values as edges in SC, not as levels of SC. An edge is a transition from logic LO to HI or from HI to LO. A rising edge is a one, and a falling edge is a zero. A sequence of synchronizing one-bits at the beginning of the message tells the receiver when to look for data-carrying edges, because it knows the rising edges in the synchronizing sequence are the data edges in these initial one-bits. Every subsequent data-carrying edge allows the receiver to adjust its expectations for when the next data-carrying edge will arrive. With a 5 MBPS bit rate, the data-carrying edges are separated by 200 ns. Each bit takes roughly 8 periods of our 40-MHz message clock. We say "roughly" because the bit rate is not exact. The transmitter's message clock is generated by a ring oscillator, and is accurate to only ±10%.
If the receiver detects an edge within one or two clock periods of another, it rejects the entire message and goes back to looking for synchronizing bits. It rejects synchronizing bits using the same constraint. The receiver tests the level of SC for premature edges twice during each bit period. In order for a random sequence of bits to create a valid message, it must satisfy both tests for all bits of the message. There are around 25 bits in the message, so the random stream must take on the correct value 50 times. Furthermore, the message must have the correct checksum at the end, which is the same as saying that it must take the correct value for another five bits.
The likelihood of a random sequence on SC matching our message protocol is roughly 0.530. To the first approximation, there are 105 opportunities for the match to occur every second, because each message occupies roughly 10 μs. We expect our bad message rate from noise and random interference to be of order one per hour. And indeed this is what we observe: after running for ten minutes we saw no bad messages with the antenna disconnected.
Most interference, however, is not random. It is generated by communication devices and its purpose is the transfer of information in a regular fashion. Such interference can have a far greater probability of creating a bad message than random interference. In a basement laboratory with −68 dBm interference power in the 902-928 MHz band, we recorded the number of bad messages in consecutive one-second intervals with a Data Recorder (A3018C). We used no Faraday enclosure, and no transmitters were running nearby.
The average number of bad messages per second was 1.1 and the standard deviation was 2.5. Given that we expect to receive of order 500 messages per second from active transmitters, this bad message rate is not significant.
A corrupted message is one that has been interfered with, but which still passes through our error-checking. A corrupted message has the same effect upon data acquisition as would a bad message with the same ID.
We can reduce the likelihood of message corruption by checking for errors in the content of the message, as opposed to only by comparing the final four bits to the first four bits. A four-bit cyclic redundancy check at the end of all transmitted messages would give us better rejection of corrupted messages. Instead of ending up as bad messages, most corrupted messages would end up as missing messages. But the cyclic redundancy check would make our Stage Four transmitters incompatible with those of Stage Three, so we decided to tolerate a higher than necessary corrupted message rate for the sake of backward-compatibility.
The text below is a list of messages between one clock message and the next, taken from data recorded from a system with six active A3013A transmitters (starting time 71.0625 s in archive M1288538199.ndf). Each line represents one message, either a clock (ID 0) or a sample (IDs 1 to 14). We give the message index, the channel number, the sample value, the timestamp, and the hexadecimal representation of the four message bytes.
24 0 7050 5 $001B8A05 25 8 42595 0 $08A66300 26 12 43431 26 $0CA9A71A 27 7 43084 31 $07A84C1F 28 10 40959 43 $0A9FFF2B 29 8 42613 53 $08A67535 30 12 405 83 $0C019553 31 7 43100 90 $07A85C5A 32 6 42185 92 $06A4C95C 33 4 180 106 $0400B46A 34 10 40987 115 $0AA01B73 35 8 42615 126 $08A6777E 36 12 43416 160 $0CA998A0 37 6 42111 160 $06A47FA0 38 7 43116 162 $07A86CA2 39 5 42234 169 $05A4FAA9 40 10 40988 177 $0AA01CB1 41 8 42661 191 $08A6A5BF 42 7 43197 218 $07A8BDDA 43 12 43330 221 $0CA942DD 44 6 42310 235 $06A546EB 45 10 41052 242 $0AA05CF2 46 8 42689 246 $08A6C1F6 47 0 7051 5 $001B8B05
These messages were recorded inside Faraday enclosures, so interference from non-telemetry sources is minimal. Message 30 is a corrupted message from No12 in which the ID has remained the same. This corruption gives rise to a glitch in the No12 signal. Message 33 has ID 4, but there is no No4 transmitter in the system. We suspect that a message from No5 was corrupted in such a way as to appear as a message from No4.
The above errors occur more often when the signal from one or more transmitters is exceptionally weak. The errors manifest themselves as spikes in the signal. Because they are rare, we can remove them with a glitch filter. More of a problem than the spikes is the poor reception that goes with the weak signals that cause spikes.
[27-JAN-17] When two transmitters send a message at the same time, we say that they collide. When two transmitters collide, we can lose one or both of their messages. Consider message detection by a single antenna amplifier, as is the case with our original Data Receiver (A3018). If the first message is much more powerful than the second message arriving slightly later, the second will be lost. The second message will become a missing message. But the first message will be received correctly. If the second message is more powerful than the first, it will interfere successfully with the first, preventing its reception. The second message may also be lost, depending upon how fast the message detector can recognize and recover from the corruption of the first message. If the two signals are of equal power, neither will be received. If we have two or more independent receiving antennas, we can hope to detect both messages. The Octal Data Receiver (A3027) provides eight receiving antennas, each with its own amplifier, demodulator, and message detetor. Collisions occur at each of the antennas, but the outcome of the collision differs from one antenna to the next, so that the rate at which we lose messages to collisions is reduced.
Collisions occur because two transmitters are transmitting at the same time. If the transmission period were exactly regular, and two clocks drifted into exactly coincidence, collisions could occur systematically at every transmission instant. But the transmission period is not regular. The average period is exact, but the moment of each individual transmission is displaced by the transmitter by a small, random, amount of time. In the A3038E, the transmission period is 64 cycles of its 32.768 kHz clock, or 1952 μs. The A3028E delays its moment of transmission by 0 to 15 clock cycles, so that the actual moment of transmission can be delayed by up to 456 μs. Or we can regard the moment of transmission as being displaced by ±240 μs about its nominal value.
We call the displacement of the transmission instant transmission scatter. We describe how the data acquisition system handles transmission scatter in our Recorder Manual. The transmitter uses the lower four bits of its sixteen-bit ADC conversion as the source of a random number. The figure below shows transmitter scatter on the oscilloscope screen.
We describe in more detail how the hardware implements the scatter in the Transmission section of our A3013 Manual. We describe the distortion of the analog signal that results from transmission scatter below.
Faraday enclosures provide isolation of transmitters from ambient interference, and also from transmitters that do not share the same receiver. If we have eight transmitters in four separate Faraday enclosures, each Faraday enclosure will have its own antenna. We combine the signals from the six antennas and feed the combination into our Data Receiver (A3018) using an Antenna Combiner (AC4A). Transmitters in these six enclosures will interfere with one another whenever their transmissions overlap.
At 512 SPS, the transmission period is 1952 μs and the transmission itself lasts for only 7 μs. Each transmitter transmits for 0.35% of the time. The chance of one 7-μs transmission overlapping another is 0.7%. The average loss due to collisions when we have n transmitters sharing the same receiver is (n-1) × 0.7%. If we have If we have twelve transmitters sharing a receiver, we will lose roughly 8% of messages to collisions when averaged over a long time period. We say the average collision loss is 8%.
The collision loss varies with time, as shown in the following graph. Its average value may be only 0.7% for two transmitters, but it's peak value can be ten times higher.
In the above graph, we see cyclic variation in reception caused by a slight difference in the transmitter clocks. Over the course of eighteen minutes, the cycles grow in amplitude from 1% to around 8% and shrink again. Another twenty minutes goes by with no evidence of collisions, and the collision cycles begin again.
The period of message transmission at 512 SPS is roughly 2 ms (64 cycles of the transmitter's 32.768 kHz oscillator). An entire collision sequence, such as the thirty-six minute sequence captured in the figure above, takes place as the two clocks drift with respect to one another by 2 ms. The ASH7KW clock we use on the A3028 is accurate to ±20 ppm over its entire temperature range. We expect differences between the clocks of 5 ppm to be common. In the example above, it takes 2200 s for the two clocks to drift apart by 2 ms. The two clocks differ by 0.9 ppm. This combination of clocks is unusual. More often we see collision cycles of period several hundred seconds, like this one taken from transmitters in live animals at ION in London.
The individual cycles in the above sequence have period thirty seconds, as we can see in the following figure.
Each cycle corresponds to a drift of 30.5 μs between the two clocks (one period 32.768 kHz). Because our clocks are only 0.9 ppm apart, we get a 34-s cycle. We see the structure of the cycle clearly. There is a 18-s period with no collisions in each cycle, and a 16-s period where the collisions take place.
The first period of collisions in the entire collision sequence begins when the earliest transmission window of one transmitter coincides with the last transmission window of the other. In our case, the two 7 μs windows overlap for 16 s (that's twice 7 μs divided by 0.9 ppm). Collisions in this first period should be rare, because only one window overlaps. The chance of a collision should be 1/16 × 1/16 = 0.4%.
As we see in the oscilloscope trace above, the four-bit values the transmitters use for random numbers are not uniformly-distributed across their sixteen possible values. Furthermore, one transmitter might be so much stronger than another that the interference is only one-way. In our example sequence, we see the first cycle has a depth 6% for No4 and 0% for No11. The peak cycles have depth 8% for both transmitters.
The second cycle of collisions occur 34 s after the first, when the clocks have drifted 30.5 μs to the next window overlap. Now the earliest window of one transmitter coincides with the second-to-last last window of the other. But now we have the second-earliest window coinciding with the last window as well. Our chance of collision is, in theory, 0.8%. The overlap lasts for 16 s, and 18 s later comes the next overlap. The chance of a collision is 1.2%. On the sixteenth cycle, all sixteen windows overlap, and the collision probability is 6%. The cycles shown in detail in the figure above are the largest in the collision sequence. Both transmitters experience a 6% drop in reception. Sixteen cycles after the peak, none of the windows overlap. The clocks drift another 1 ms apart over the next 1200 s and there are no collisions. Now the entire sequence starts again.
If we place n transmitters in the same Faraday enclosure, it is inevitable that their sixteen transmission windows will coincide at some point in time. At that time, the collision rate between transmitters will be at its greatest, and reception will be at a minimum. If we consider a message occurring in one of the windows, the probability of no other message occurring in the same window is 0.94(n−1). If we assume near-perfect reception in the absence of collisions, which appears to be the case in Faraday enclosures, this probability is the minimum reception rate for the n transmitters.
| Number | Average (%) | Minimum (%) |
|---|---|---|
| 1 | 100.0 | 100 |
| 2 | 99.3 | 94 |
| 3 | 98.6 | 88 |
| 4 | 97.9 | 83 |
| 5 | 97.2 | 78 |
| 6 | 96.5 | 73 |
| 7 | 95.8 | 69 |
| 8 | 95.1 | 65 |
| 9 | 94.4 | 61 |
| 10 | 93.7 | 57 |
| 11 | 93.0 | 54 |
| 12 | 92.3 | 51 |
| 13 | 91.6 | 48 |
| 14 | 90.9 | 45 |
For robust reception we need to receive more than 80% of messages for 95% of the time (robustness of 95% or higher). It's not obvious from the minimum and average reception values whether reception will be robust. To obtain a good estimate of robustness, we simulated the collision cycles of n transmitters. Our simulation program, collisions_1.pas, simulates sets of n transmitters working together with randomly-distributed clock periods over a period of two thousand seconds. The figure below shows reception from the first four transmitters in a set of fourteen.
When the simulation begins, all clocks are synchronous, and we obtain the minimum reception. After that, the interactions between the clocks become complex. For a 100-s detail, see here. For each simulation of n transmitters, we obtained n values for minimum reception, average reception, and robustness. These values were consistent from one transmitter to the next to within a few percent, so the values we give in the table below are the average values taken over the n transmitters.
| Number | Average (%) | Minimum (%) | Robustness (%) |
|---|---|---|---|
| 1 | 100.0 | 100.0 | 100.0 |
| 2 | 99.2 | 91 | 100.0 |
| 3 | 98.0 | 87 | 100.0 |
| 4 | 97.8 | 84 | 100.0 |
| 5 | 96.2 | 79 | 100.0 |
| 6 | 95.6 | 72 | 99.8 |
| 7 | 95.2 | 72 | 99.8 |
| 8 | 94.4 | 68 | 99.6 |
| 9 | 94.0 | 62 | 99.3 |
| 10 | 92.7 | 57 | 98.5 |
| 11 | 92.2 | 58 | 98.2 |
| 12 | 91.5 | 52 | 97.3 |
| 13 | 90.7 | 52 | 89.0 |
| 14 | 89.9 | 50 | 81.7 |
Robustness is greater than 95% all the way up to n = 12, so the collision tolerance of a set of 512 SPS transmitters is 12. This is why we talk about 12 transmitters sharing a single receiver in other sections. Robustness for n ≤ 5 is 100%.
In our simulation, we assume that any collision between two transmitters will result in the loss of both messages. This is not true in practice. If the power received from one transmitter is 12 dB greater (16 times greater) than the power received from another, the more powerful signal will dominate. If reception of the more powerful signal is taking place, the weaker signal will be ignored. If the weaker signal is being received, there is a good chance that the receiver will have time to abandon the weaker message and receive the stronger message. In our recording of two stationary transmitters, shown here, we see that fewer messages are lost from No7 than No4. Transmitter No7 is in air with an 80 mm antenna. Transmitter No4 is in water with a 50-mm antenna. We assume the signal from No7 is stronger than that from No4.
The following graph shows measured transmission over the course of half an hour with nine active transmitters.
Average reception over the half-hour period was 92% for No3 and 99% for No1, with the others in between. The minimum reception observed in any four-second period was 71% from No2 and 94% for No1, with the others in between. Reception is almost always greater than 80% for all transmitters, so we conclude that we obtain robust reception from nine transmitters despite collisions.
All these calculations assume that we have only one antenna amplifier and demodulator with which to detect all transmitted messages. But the Telemetry Control Box (TCB) provides sixteen independent antennas. When two transmitters collide but are in different locations, both messages are likely to be received correctly because at each location one transmitter dominates over the other. We placed nine transmitters of various sample rates in a Faraday enclosure, emitting a total of 8192 SPS, and received their messages with three independent antennas. We observe an average loss of 4%. Our simulation suggests a loss of 12% at each of the antenna. When we place 12 of A3038L in an FE3A enclosure with four independent antennas, each transmitter emits 2048 SPS, so we have the equivalent of 44 transmitters of 512 SPS. Average loss is 10%.
We define operating range as the greatest range from the pick-up antenna at which we obtain robust reception. An A3019A transmits 512 messages per second, so the operating range is the greatest range at which we receive 410 or more messages per second in 95% of orientations and positions at the operating range. In practice, we operate the transmitters inside a Faraday enclosure, so their operating range must be adequate to cover the range from the antenna in the center of the enclosure to the farthest corner of the enclosure volume.
When we hold a transmitter between two fingers in our basement laboratory, and move it around randomly at range 100 cm for a minute (see movie), we receive 98% of the messages it transmits. The operating range of the A3019 is greater than 100 cm in our basement laboratory. In our our office in the center of Waltham, operating range is closer to 70 cm. Operating range in the ION laboratory in London appears to be around 30 cm. The decreasing operating range is the result of ambient interference, which dominates transmitter messages whenever the transmitter signal is attenuated by reflections and unfavorable orientation.
Regardless of the operating range of transmitters in a laboratory, we recommend the use of enclosures with conducting walls to block out ambient interference. Such enclosures are called Faraday cages. We call them Faraday enclosures so we don't get them confused with animal cages, which are used to contain animals. We discuss ambient interference below, and we describe the performance and construction of Faraday enclosures in our Faraday Enclosures report.
Because Faraday enclosures give us at least a 30-dB (one thousand-fold) reduction in ambient interference, they increase the operating range of our transmitters by a factor of 30 (square root of one thousand). Even if the operating range is only 10 cm without a Faraday enclosure, it will be 300 cm within a Faraday enclosure. We expect robust reception from transmitters operating within an enclosure 500 cm × 500 cm × 200 cm, provided the antenna is in the center of the floor of the enclosure. Our FE2A enclosure is far smaller than this.
Robust reception within a Faraday enclosure appears to be guaranteed. Nevertheless, we always benefit from increased signal strength compared to ambient interference, and we have worked hard to increase the operating range of our transmitters in the absence of Faraday enclosures. We devote the remainder of this section to a discussion of what phenomena limit the operating range of our transmitters. The discussion explains why we use a poorly-tuned loop antenna to receive our transmitter signals, and why we use a poorly-tuned bent antenna to transmit them.
When we implant a transmitter in an animal, the 50-mm antenna of an A3013A-E will resonate more efficiently, and so transmit more power (see Antenna Length). We may lose some or all of that additional power by absorption in the animal's body (see Transmitter in Baby Rat Corpse).
We compared the operating range of the Data Receiver (A3010B), which is the Stage Three receiver upgraded with new firmware and an antenna socket, and the Data Receiver (A3018A), which is the original Stage Four receiver. (The original Stage Three receiver, the A3010A, suffered from a firmware bug that reduced its performance even farther with weak signals.) We used the same Subcutaneous Transmitter (A3013A) for both receivers. We used the same Loop Antenna (A3015A) for both receivers. We unplugged it from one receiver and plugging it into the other.
It appears that our Stage Four development has doubled the effective range of the transmitters. We claim that we are nearing the physical limits of message detection, as we shall now explain.
The noise in our receiver is the thermal and amplifier noise at the antenna input. The interference is RF power in the receiver's pass-band arriving at the antenna from transmitters other than those the receiver is intended to receive. The effective noise power at the input of our demodulating receiver is −90 dBm (see Noise and Interference). But interference power, even in our basement laboratory, is over a hundred times more powerful than this noise, at −68 dBm (see Noise and Interference).
Our signal must have power 12 dB greater than the interference in order to avoid corruption (see Foreign Interference). When we transmit power across 1 m of space from a quarter-wave antenna to a loop antenna, we lose at least 32 dB compared to connecting the power directly to the receiver circuit with a cable (see Reception). Even with our omni-directional antennas, an unfavorable relative orientation of the transmitting and receiving antennas causes a 17 dB drop in received power (see Omnidirectional Antennas and Transmit Antenna). We must also deal with reflections, or multi-path interference (see Multi-Path Interference and Radiated Power). Reflections of the main signal interfere at the receiver. We can easily get a 10-dB loss due to destructive interference. In other words: we can lose 90% of our power easily
Our transmitters produce roughly −4 dBm (see Radiated Power and Modulating Transmitter). Transmission across 1 m of air with can drop our received power to −53 dBm. This −53 dBm is still more than 12 dB above our interference power of −68 dBm.
But we must contend with multi-path interference as well, which give rise to reception dead spots. With the antenna in an unfavorable orientation, in which we receive only −53 dB from line-of-sight transmission, we have power radiating more effectively in other directions. This power can reflect off nearby conducting surfaces and arrive at the receiving antenna with as much strength as the line-of-sight signal. The reflection adds to the line-of-sight signal. It might reinforce the line-of-sight signal, or cancel it, depending upon their relative phase.
The phase difference between a reflected and line-of-sight wave is a strong function of frequency. If the reflected path length is three meters long, this is a hundred wavelengths. A 1% change in frequency will cause a 2π change in phase. A 0.25% change in frequency will cause a π/2 change in phase. The figure below shows two holes in the response of our our Demodulating Receiver (A3017). The antennas are 1 m apart, on either side of our body.
We see that each hole is about 1 MHz wide. A 0.1% change in frequency causes the cancellation to stop. We take such sharp holes in the frequency response to be evidence of multi-path interference. The width of the holes is consistent with a reflected path length of several meters. Longer path lengths would give holes that were even more sharp.
Aside: The two holes are separated by 24 MHz, and you will notice yet another hole, on the left side, which is 24 MHz below the middle hole. As the frequency changes by 24 MHz, the phase difference between the line-of-sight and reflected waves changes by 2π. The number of wavelengths in the path difference changes by one. Because the frequency changes by 2.5% to bring about this one-wavelength change, the path difference must be 40 wavelengths, or 12 m. This suggest to us that the wave is bouncing off one of the metal shelves five or six meters from our transmitting antenna. If that's the case, then the reflected signal will be about 25 dB weaker than the line-of-sight signal, except for the fact that our body is in the line of sight. Human tissue attenuates 900 MHz by approximately 1 dB/cm, so we expect a loss of around 25 dB through a human torso. Both signals are each strong enough for reception, but they cancel one another at particular frequencies.
With 10 dB loss due to cancellation by reflections occurring at the same time as 17 dB loss due to poor antenna orientation and 32 dB loss due to transmission across 1 m of air, our signal drops to −63 dBm, which is only 5 dB above our −68 dBm interference. Reception will fail.
With a favorable orientation of the transmitter, however, we can expect −36 dBm at 1 m, and − 56 dBm at 10 m. Even at 10 m, we can receive a signal that is 12 dB above our interference. In fact, we find we can get reliable reception at up to 15 m in our lab by orienting the transmitter properly.
Radio frequency power from sources other than our subcutaneous transmitters is what we call ambient interference. If it's large enough, ambient interference can dominate the transmitter signals and cause missing messages. In early 2009, we determined that our operating range in the ION animal laboratory in London was only 25 cm, after observing a 50-cm operating range for most of 2008 (details in this e-mail). A range of 25 cm is not adequate to cover even a single animal cage, let alone four at once. We assumed that ambient interference was to blame for the the short operating range, and began to look at ways to block ambient interference from reaching our receiving antenna.
We can block out ambient interference with a Faraday enclosures, which is a box with electrically conducting walls. There can be holes in the conducting walls, but the holes must be less than 1% of the wavelength of the radiation if we want to block 99% of the power. In our case, the wavelength in air is around 300 mm, so holes less than 3 mm will be fine. We describe our experiments with home-made Faraday enclosures in our Faraday Enclosures report.
Another way to block ambient interference is to turn your entire laboratory into a Faraday enclosure. You can paint the walls, ceiling, and floors with shielding paint. You could cover the windows with aluminum mosquito mesh, or make curtains out of shielding fabric. Alternatively way to block ambient interference is to move your animal laboratory into a basement with a concrete ceiling.
Our FE2B Faraday enclosure sells for around a thousand dollars US. It is large enough to contain two rat cages. Four litter-mates could live together in each cage, each with an implanted transmitter. We believe this is the most cost-effective way of performing long-term studies.
Ambient interference can penetrate the recording system through the cable that joins the receiver to the LWDAQ Driver. To cut down on such penetration, we must use a shielded LWDAQ cable for this connection, as we describe in Shielding.
[19-JUL-23] Our transmitters provide high-fidelity EEG recording. The figure below is an example of the frequency response graphs we obtain during quality control of all batches of transmitters we manufacture. You will find a bunch of example frequency response plots here.
When we drop a transmitter in water, we are able to measure the noise on its biometric inputs. For our standard amplifier with gain ×100 and input impedance 10 MΩ, this noise is 4-6 μV rms, referred to the analog input. With gain ×10 the noise increases to aroun 12 μV rms.
The distortion of a signal by our telemetry system is the extent to which it changes the shape of a signal. We apply a 10 mVpp sinusoid to the X and Y inputs of an A3049AV3. The AV3 is equipped with two 160-Hz amplifiers. Input dynamic range is 27 mV. We increase the frequency from 1/8 Hz to 200 Hz. For each frequency, we obtain the spectrum of the signal and measure the power outside the sinusoidal frequency as a fraction of the sinusoidal power using this script. We express the result in parts per million.
The distortion of the X is dominated by random electronic noise. There are no significant peaks in the spectrum outside the fundamenta.
We note that the distortion generated by the new A3047, A3048, ajnd A3049 transmitters is hundreds of time less powerful than that of their predecessors, the A3013, A3019, and A3028. The new transmitters sample their signals uniformly, thus eliminating the scatter noise present in earlier devices.
[01-JUL-22] We encapsulate transmitters in epoxy and coat them in silicone. For an account of our initial work on rugged encapsulation that is resistant to water, fatigue, and vacuum, see our here. Our encapsulation procedure does not use molds. We dip the circuits in epoxy while in a vacuum, then allow air into the vacuum to force the epoxy in and around all components. We rotate the transmitter while the epoxy cures, and later dip several times in a medical-grade silicone dispersion to provide a resiliant outer coating. Encapsulating by dipping produces a far smaller device, and we can adapt our procedure easily to new shapes. The disadvantage of epoxy and silicone is that they are permeable to water vapor, so that it is possible for water to condense within the circuit. In the warm body of an animal, condensation causes corrosion. We select roughly one in ten of the transmitters we manufacture and subject them to accelerated aging in hot water, where they run for weeks or months until corrosion stops them from functioning. We take them out every other day and handle them and check that they are functioning correctly. The OSI product warantee guarantees our implantable devices against corrosion for their minimum operating life or for one year after we ship them, whichever is greater.
Animals will sometimes scratch at stitches and incisions. When they scratch so vigorously as to penetrate their own skin, their claws tear at the silicone coating of their implanted SCT. Once the silicone is breached, water penetrates to the epoxy coating beneath the silicone. Current can flow from the battery terminal, through a breech in the silicone, to the reference electrode of the SCT, thus completing a circuit that causes corrosion on the battery surface. After a week we will see brown and green oxide beneath the silicone.
The current that flows through the breech in the silicone will be of order a few microamperes. The transmitter battery will drain slighly more quickly. In the case of a transmitter like the one shown above, operating life may drop from 40 days to 35 days. But the breech will corrupt the signal recorded by the transmitter. As the animal moves, the breech will flex, changing the current that flows through it, and generating movement artifact of several hundred microvolts in the recorded signal. The generation of this artifact is so reliable that we use its generation to check for flaws in the coatings of newly-made transmitters. We soak the transmitters in water for several days, then transfer them to hot water, and look at the signals they transmit, watching for the steps and swings that reveal a silicone breech.
The silicone coating on our transmitters is roughly 0.5 mm thick. We want the silicone to be tough enough to endure handling, dropping on the floor, implanting, and explanting without the silicone tearing or cracking. The thicker we make the silicone, the tougher the coating will be. But the ticker the silicone, the greater the volume of the transmitter. The larger the transmitter, the more likely it is to irritate its host animal. Once an animal is so irritated as to scratch open an incision, the study of that animal is over. Thus, we do not equip our implants with silicone thick enough to survive being scratched repeatedly, let alone bitten.
Our standard encapsulation involves several coats MED-6607 unrestricted medical grade silicone. The humidity and air flow around the transmitter while the silicone is curing is critical to the uniformity and clarity of its appearance, but less critical to its performance as a coating. If something goes wrong with climate control in our curing chamber, one of the layers, usually the first layer, will wrinkle, and the result is an ugly transmitter like the one shown below.
We have poached dozens of transmitters like the one shown above in water at 60°C to test their corrosion resistance. The ugly transmitters are just as tough and long-lived as the good-looking transmitters. Nevertheless, we won't ship ugly transmitters without warning, and we will hold them back and replace them if our customer requests us to do so. They carry the same one-year warranty as all our implantable devices.
[29-NOV-23] Our electrode leads are a flexible helix of stainless steel wire insulated in silicone. Standard leads are 20-130 mm long with an accuracy of ±3 mm. We can make longer leads by joining shorter leads, but we charge extra for doing so. We describe how we arrived at this design in Flexible Wires.
| Lead Code |
Outer Diameter (mm) |
Spring Diameter (μm) |
Wire Diameter (μm) |
Resistance (Ω/cm) | Names | Comment |
|---|---|---|---|---|---|---|
| A | 1.0±0.2 | 450 | 100 | 6.3 | Thick Lead | Discontinued, too thick. |
| B | 0.7±0.1 | 450 | 100 | 6.3 | Thin Lead | Up to 280 mm long. |
| C | 0.5±0.1 | 250 | 50 | 25 | Very Thin Lead | Up to 130 mm long. |
| D | 0.8±0.1 | 500 | 150 | 1.6 | Stimulator Lead | Up to 130 mm long |
We order our 450-μm diameter springs in 300-mm sections. We order our 250-μm leads in 150-mm sections. We can manufacture 0.7-mm diameter insulated leads up to 280 mm long and 0.5-mm diameter insulated leads up to 130 mm. The 0.5-mm diameter leads are far more flexible than the 0.7-mm leads. They are less likely to cause irritation and infection in the subject animal. But the spring in the 0.5-mm leads is delicate. The wire in the 0.5-mm lead is half the diameter of the wire in the 0.7-mm lead. The spring itself is one quarter as strong. They will survive the fatigue of animal movement, but they are easy to damage with a scalpel during implantation or extraction. Removing insulation from a 0.5-mm lead is a delicate operation. We cannot provide screw or pin terminations on 0.5-mm leads because the wire breaks so easily at the edge of the solder joint. We can, however, solder the 0.5-mm leads to X-Electrodes directly and insulate with silicone afterwards, which is how we make the EIF-XAAX electrode interface. The D-Lead is for use with implantable light-emitting diodes (ILEDs). It's wire is thicker and its spring pitch is greater, so that its resistance is less than one quarter that of the B-Lead. The D-Lead is stiffer, but it makes it possible for us to deliver tens of milliamps to implantable lamps in rats, when the resistance of B-Leads would make such currents impossible.
[27-NOV-23] We provide a variety of terminations for our electrode leads, and a variety of depth electrodes to which these terminations can be connected. See our Electrodes Catalog for a list of terminations and depth electrodes with links to photographs and drawings.
[14-NOV-24] The table below lists the various antennas we use with our 915-MHz telemetry system. They vary in length, material, and shape. We will recommend an antenna based upon your animal mass, implant operating life, and implant type.
| Antenna Code |
Length (mm) | Description |
|---|---|---|
| A | 50 | Stranded steel loop antenna for rats 360-μm diameter 7×7 304SS wire insulated in clear MED-6607 silicone. |
| B | 30 | Stranded steel loop antenna for mice, 360-μm diameter 7×7 304SS wire insulated in clear MED-6607 silicone. |
| C | 13 | Straight antenna of helical wire for pups, 450-μm diameter 316SS helix insulated in clear MED-6607 silicone. |
| D | 30 | Stranded steel loop antenna for small mice 250-μm diameter 7×7 304SS wire insulated in clear MED-6607 silicone. |
| E | 50 | Stranded steel loop antenna for mice or rats 250-μm diameter 7×7 304SS wire insulated in clear MED-6607 silicone. |
The 50-mm A and E antennas produce the strongest signal when implanted in rats. The 30-mm B and D antennas fit easily into mice without folding. The D and E antennas are made with a thinner stranded wire. We can fold the 30-mm D antenna through 45 degress with a 1-g force, while the 30-mm B antenna requires a 4-g force. Because of their flexibility, the D and E antennas are a natural choice for mice and rats respectively. The A and B antennas are, however, more resistant to fatigue. For implantations longer than three months, we recommend the A and B antennas. We no longer make the C-Antenna because it transmits ten times less power than the loop antennas, while the D-Antenna is just as easily accommodated by a small animal as the C-Antenna.
[05-FEB-20] When we implant a transmitter and its leads in an animal body, there exists a capacitance between the transmitter circuit and the animal body, and also a capacitance between the lead wires and the animal body. The capacitance between the circuit and the animal body has for its dielectric the epoxy and silicone used to encapsulate the transmitter. This capacitance is of order 10 pF. The capacitance between the lead wires and the animal body has for its dielectric the silicone insulation of the leads. This capacitance is of order 10 pF/cm, and therefore dominates the total capacitance between the transmitter and the animal body. We show how we estimate and measure these capacitances in the Body Capacitance section of one of our early transmitter manuals.
[29-JUN-18] The active current consumption of a transmitter increases linearly with the number of samples it transmits per second. Divide the capacity of the battery by the active current conusmption to get the maximum number of hours of operation the device can provide.
The operating life of a transmitter is how long it takes to consume its battery capacity in its active state. The A3028E provides 1 voltage-sensitive input (for EEG), 1 magnetic field input (for turning the transmitter on and off), and one radio-frequency output (for transmitting messages to the receiver). During radio-frequency transmission, the A3028B consumes around 10 mA, which would run its battery down in one day. But transmission takes place only 1% of the time, so current consumption is reduced by a factor of one hundred. The transmitter trusts that the receiver will be listening all the time, and with such vigor as to pick out messages from a dozen different from a continuous, random message background. The transmitter consumes 75 μA. The receiver consumes 20 W. We have moved the complexity and power consumption of communication into the receiver, out of the transmitter, and this is why our transmitters last for hundreds of times longer than any other radio-frequency telemetry devices on the market. These other radio-frequency systems use transceiver chips that set up continuous reception and transmission between the implanted device and its receiver.
The shelf life of a transmitter is how long it takes for the transmitter to consume its battery capacity in its inactive state. When a transmitter is inactive, its magnetic switch consumes some current. In the A3028A, the switch consumes around 1.5 μA. In the A3028P it consumes around 0.8 μA. The A3028A is equipped with a BR1225 with capacity 48 mA-hr. These cells last for ten years or more without losing charge. At 1.5 μA, the battery will run down in about three and a half years.
[09-FEB-24] All our implantable devices are equipped with non-rechargeable, lithium primary cells. We experimented with rechargeable implants for several years and found them to be unreliable, toxic, and vulnerable to corrosion. They are unreliable becasue we cannot guarantee that our customers will connect the charging circuit correctly, nor that they will check the state of the battery before re-implantation. They are toxic because they emit gases when they are discharged. They are vulnerable to corrosion because the battery expands and contracts, which cracks its epoxy encapsulation, creating cavities in which water can condense. Our collaborators lost many animals to unresponsive rechargeable transmitters. Furthermore, rechargeable cells provide one third the charge capacity per unit volume of non-rechargeable cells. If we want to re-use transmitters because we absolutely do not want to buy new implantable transmitters for each experiment, we can deploy a Head-Mounting Transmitter (A3040, HMT). The HMT mounts on an animal using a connector that is cemented permanently to the skull. We can remove the HMT and replace its coin cell battery in a few minutes. We sell HMTs with a one-year warranty against unfortunate accidents that destroy the device.
[29-JUN-18] Faraday enclosures ensure robust reception from implanted transmitters. They also make sure that the telemetry system conforms to all local radio frequency regulations, because the enclosure keeps our telemetry signals contained within its walls. We describe the development of our enclosures in Faraday Enclosures. A single FE3A bench-top enclosure, with a transparent shelf installed, accommodates six animal cages and provide recording from at least forty cohabiting animals. A single FE5A canopy enclosure accommodates an entire rack of eighty individually-ventilated cages. Not only is radio-frequency interference kept out of the enclosure, but so is low-frequency electrical noise such as mains hum. Conversely, the transmitter signals are confined within the enclosure. The signal power outside the enclosure is too weak for detection by standard instruments, which guarantees that the system violates no local radio-frequency transmission rules. Furthermore, we can operate multiple recording systems in the same room, because their signals will not interfere with one another.
[25-JUN-19] We may want to isolate our animals from sounds generated outside our recording system. According to our measurements, the steel mesh walls of our faraday enclosures do not provide attenuition of sound in the range 200 Hz to 40 kHz. Clear vinyl sheet, however, is an excellent absorber of the ultrasonic frequencies audible to rats and mice. A curtain of overlapping vertical strips of vinyl provides a factor of one hundred reduction in the power of ultrasonic waves, while at the same time allowing ventilation.
Acknowledgement: Every detail of implantation in this section comes directly from our customers. We thank them for answering our questions. We do not mention our customers by name in our public documents, out of respect for their privacy.
[22-MAR-24] A transmitter and its leads must fit comfortably in an animal if the implantation is to be a success. In the experience of our customers, implanting transmitters in mice and rats of all sizes, we obtain the following rule of thumb: a mouse or rat can tolerate a single implant up to 0.07 ml per gram of its body weight, or two implants of 0.05 ml/g each. Assuming a density of 1.8 g/ml, which the approximate density of our A3049 devices, a mouse or rat can tolerate a single implant up to 0.13 g/g or two implants up to 0.09 g/g. For an independent assessment of what mice and rats can tolerate, consider the implantation of mini-pumps, such as those made by Alzet. According to Alzet, a 0.5-ml pump is accommodated by a mouse 10 g and larger, 1.0-ml is accommodated by 20 g and larger, and the 6.5-ml is accommodated by rats of 150 g and larger. Our veterinary collaborators tell us that two such pumps can be implanted in one animal without excessive discomfort.
A four-week old rat weighs roughly 80 g. According to one paper, rats at age four weeks and can tolerate an implant of 5.6 ml. In our experience, a 2.8-ml A3028E is well-tolerated by an 80-g rat. But we note that an 80-g rat is growing quickly: within two weeks it weighs 150 g. Our A3028L is 6.0-ml device of mass 13 g. According to one implanter, "The A3028L is probably not well tolerated by smaller rats. When I implant in rats under 300g, if the transmitter is placed too posterior, over the femoral region it can be quite irritating. I have had two rats that scratched through the skin in this region." Male Sprague-Dawley rats reach 300 g at 8 weeks, while females reach 300 g only after 16 weeks.
Most strains mice are roughly 10 g after 3 weeks and 20 g after 6 weeks. In our experience, a 20-g mouse is comfortable with a 1.3-ml A3049A, but a 10-g mouse barely tolerates the same device. We have obtained reliable recordings from rat pups only four days old using a 1.4-ml transmitter, but a better choice today would be our newer, single-channel A3048P 0.8-ml, 1.2 g transmitter.
One of our collaborators measured the weight of four 28-day-old rats before and after implantation of a 1.4-ml, 2.6-g transmitter. The animals weighed as little as 55 g at the time of implantation. Transmitter volume was 1.4 ml not including leads, and mass was 2.4 g including leads. A control animal received no transmitter and was subject to no surgery. All five animals cohabited in the same cage in the week following, and we recorded EEG continuously from the four implanted transmitters.
Weight gain following surgery is presented in the following table. See also Chang et al. for implantation procedure and EEG recordings.
The video of the freely-moving animals, and their weight gain after surgery, suggests that a 1.4-ml implant in a 55 g animal has no significant impact upon its well-being. We do not have such weight-gain measurements for 20-g mice implanted with our 1.4-ml transmitters, but Wright et al. describe how mice of initial weight 18-22 g tolerated 1.4-ml for at least three weeks to the satisfaction of UK veterinary inspectors.
We describe implantation procedures in detail in the supplementary materials of Chang et al. 2016, the first section of Wright et al. 2015, and in our original methods paper Change et al. 2011. In the text below you will find the latest details and advances in the implantation procedure. Each implanter has their own preferred method, so we supply here a collection of advice and suggestions collected from a variety of methods. We have a private collection of implantation videos that you may also find useful. These videos are, however, available only to our active customers, as agreed with the implanters who provided the videos.
There are two established procedures for implantation. The two-incision procedure uses one incision in the back or side of the animal to accommodate the body of the transmitter, and a second incision in the scalp to give access to the skull. We tunnel the electrode leads under the skin to the skull. On the skull, we insert screws or wires into skull holes and fasten them in place with dental cement. Or we insert pins into depth electrodes, lower the electrodes into the brain, and secure with dental cement. The two-incision procedure works in all animals.
The single-incision procedure uses one incision in the scalp large enough to slide the body of the transmitter down under the neck to the abdomen. This single-incision procedure works better in mice than in rats, because the distance the transmitter must be thrust under the skin is greater in rats.
In both procedurs, we have the option of closing the incision in the scalp, or building up a head fixture of dental cement, and fastening the edges of the incision to the head fixture with n-butyl cyanoacrylate. We recommend 3M RelyX Unicem 2 for dental cement and Vetbond for cyanoacrylate. Closing the incision can create tension and irritation that leads to scratching and opening of the sutures. Leaving the incision open invites infection at the interface between the skin and the dental cement.
When implanting, you may be working on steel tables with steel implements. These can become magnetic and they can turn on and off the transmitter while you are working. One way a table or a tool can become magnetic is by storing your on-off magnet on the table or the tool handle. Turning off and on the transmitter during implantation causes no harm to the transmitter, but can cause concern that the transmitter has failed when it has not. Implant the SCT with the battery facing in, so the circuit board holding the magnetic sensor is closest to the skin. Use a large magnet. The field of a small magnet will not penetrate easily through the skin.
The choice of termination at the tip of the leads depends upon what we want to measure, as well as the size of the animal. We present our work on electrodes here, and you will find a list of currently-available terminations and electrodes here. The most important consideration when implanting an electrode is that the conducting tip of the electrode must be fixed in place with respect to the body tissue as the animal is moving. If the electrode moves with respect to the brain or skull tissue, we will see step artifacts in our EEG recording. Our objective is to reduce the number of step artifacts we see from moving animals to the point where their frequency is has no significant impact upon our automated analysis of the signal. Our Event Classifier will be fooled by no more than 5% of step artifacts, so if we want the classifier to make mistake no more than one such artifact per day for an actual EEG event, we must reduce the frequency of such steps to fewer than one per hour.
Our first successful EEG electrodes were a stainless steel screw soldered to the end of the electrode lead. In mice, the skull is only 200 μm thick near the bregma. For EEG monitoring, we recommend a 0.6-mm long, 0.5-mm diameter threaded electrode. In rats, the skull is around 500 μm thick, and over the course of many weeks, will grow thicker. The screw we recommend for rats is 3.2-mm long and 1.5-mm in diameter. This screw is large enough to accommodate the growth of a young adult rat from 50 g to 200 g and still maintain good contact with the brain for consistent EEG amplitude throughout months of EEG recording. Nevertheless, soldered screws have the following disadvantages.
We no longer recommend soldered screws as EEG electrodes, although we will supply them when asked. Our preferred EEG electrode is a bare wire held in a skull hole by a screw. The tip of the bare wire touches the surface of the brain, or enters the brain by a fraction of a millimeter. With these bare-wire electrodes, we obtain larger amplitude EEG. Provided we take care during construction of the head fixture, the wire will be held firmly in place and we will see fewer than one step artifact per hour. Cutting the leads to the correct length, drying the skull, preparing the skull with n-butyl cyanoacrylate, covering all exposed metal with dental cement, and making sure the dental cement has time to cure before closing the incision, ensure the bare wire is stationary with respect to the skull as the animal moves around.
When we plan to use bare-wire electrodes, we can cut the leads to the optimal length during implantation. We want to avoid one lead being subjected to more tension than another. If we have three leads running from the transmitter to the skull, one electrode might be 10 mm farther from the transmitter than another. We make its lead 10 mm longer. Now we can run the leads together in a bundle from the skull to the transmitter and no one lead will be pulled out of the bundle because it is too short, nor pop out of the bundle because it is too long. Having cut the leads to length, we must expose some bare wire at the newly-cut tip of the lead. When working with our 1.0-mm or 0.7-mm diameter leads, we can grab the exposed tip of the 100-μm diameter stainless steel wire and pull some wire out. We straighten with forceps, insert a bend for the skull hole, and trim for the desired depth of penetration from the skull surface. When working with our 0.5-mm diameter leads the wire is too fragile to extract in this way. We must cut through the insulation near the tip with a blunt scalpel and use two pairs of tweezers to pull the free end of the insulation away from the main body of the lead. This exposes a stretched length of 50-μm diameter wire that we can straighten, bend, and trim. For more on insulation removal see Solder Joints.
Here is a suggestion from one implanter of bare wire electrodes. "Make sure neither screw is anywhere near muscles and that the screws and about half a centimeter of wire coming from the exposed lead is also covered by dental cement so that neither moves at all. I also make completely sure that the cement is very hard before suturing the back incision where the transmitter is so that there is no chance of anything moving before the cement dries." In recent recordings, a well-secured pair of electrodes produced zero step artifacts in ten hours of EEG we analyzed, while a poorly-secured electrode produced several artifacts a minute. One recording is easy to search through automatically for spikes and seizures. The other will be almost impossible to work with.
Rats and mice scratch at incisions and other points of irritation. The electrode leads of the transmitter, running from the back to the skull beneath the skin of the neck, will irritate the animal if they are poorly routed or if they are too rigid or thick. The diagram below shows us how not to route the wires in a rat, and how best to route the wires in a rat.
When the wires were routed directly up the top side of the neck, half the rats scratched the leads out of their skin, which is an indication of intense discomfort on their part. When the wires were routed along the side of the neck, with enough slack, none of the rats scratched the leads out.
If we want to inject an agent into the brain, and record EEG near the site of the injection, we can use a guide cannula to hold the bare wire in place, provided we insulate the wire from the steel shaft of the guide cannula with dental cement. If the steel wire touches the guide cannula, the EEG amplitude will drop because the steel guide has low electrode impedance. To provide insulation, we cover the bare wire with superglue, cut the end off to expose steel at the tip, bend by 90° and insert in the skull hole. We lower the guide cannula into the same hole, but separated by a few hundred microns from the bare wire, so they do not touch. The following diagram attempts to explain the arrangement.
The A3049 and A3047 SCTs come in versions that provide two or more channels each with their own reference, so that we can record potentials in different parts of the body. To record EEG, EMG, and ECG, for example, we need a three-channel transmitter with six leads.
To obtain the electrocardiogram (ECG) recording shown above, our collaborators developed the following method. Cut the ECG leads to the correct length, stretched the final 10 mm each lead, and cut around the insulation 5 mm from the tip. Once cut, the insulation pulls away from a 1-mm length of wire. They cover the tip with a silicone cap, which they hold in place with a 0/5 silk suture by squeezing the tip onto the lead. During surgery, they tie the exposed wire to the thoracic muscle with 0/5 sutures. One wire on one side of the heart, the other diagonally opposite.
To record electrogastrogram (EGG), we use the same method, suturing the two electrodes to the gut. To record electromyogram (EMG), two methods are in use among our customers. One uses an incision and adhesive. We prepare 1-mm of bare steel helix at the end of each EMG lead. We make a 1-mm wide opening between muscle fibers in two places on the muscle. We use cyanoacrylate (superglue or vetbond) to fasten a helix into each slit. Cyanoacrylate bonds well to all animal tissue and to metal. The glue will hold the two coils in place between the muscle fibers. Another method is to secure a 1-mm helix to the exterior of the muscle with a suture, such as Ethicon Sutures Vicryl 5/0.
[13-MAR-24] If we end our recording before we exhaust the battery of a transmitter, we have the opportunity to remove our telemetry sensor from our subject animal, clean the sensor, refurbish the ends of the leads, and implant again. We must take care when cutting the device out of the animal, especially if the existing implantation is more than a week old. Scar tissue builds up around the transmitter, holding in place, and wrapping around the antenna and leads. The leads are brightly-colored, but the antenna can be hard to see. The lead tips may be bound in place with dental cement. We can cut the leads where they enter the cement, then refurbish the lead tips by removing silicone to expose the bare steel wire, as we describe in Insulation Removal. If we are want to attach a screw or pin to the steel wire, we will need acid flux and a hot soldering iron, as we describe in Solder Joints. Attaching our own pins and screws is hard. If we would rather save the existing pins and screws, we must remove the cement with the lead ends embedded. To remove the cement, we and soak the leads and sensor in 20°C acetone for six hours. The cement will dissolve. The silicone of the sensor and leads will absorb acetone, but not so much as to cause significant damage to the structure of the silicone. As we describe in Silicone and Solvents, we must wash twice with clean acetone after dissolving the cement, so as to remove all traces of cement residue from the silicone surface. Once the sensor and leads are clean, leave them in air for twenty-four hours so that the acetone held by the silicone will evaporate.
[19-DEC-23] The cause of failure for implanted transmitters is corrosion in the 100% humidity of the animal's body. We encapsulate our transmitters in epoxy. Epoxy is water-proof, but it is permeable to water vapor. Once water vapor arrives at a cavity within the epoxy, it will condense and cause corrosion. Our encapsulation process is designed to eliminate cavities. Even if there are no cavites, the electrical components themselves can corrode. Ceramic capacitors in particular are prone to cracking during assembly and handling. In the warmth and humidity of an animal's body, these cracks will corrode until they cause a short-circuit between the capacitor plates.
We measure the corrosion resistance of our transmitters by means of accelerated aging. For every ten transmitters we ship, we set aside at least one extra, turn it on, and place it in water in our oven at 60°C. Every day we take the transmitter out and check how it is doing. According to statistical mechanics, the rate at which a chemical reaction proceeds is proportional to e−E/kT, where T is absolute temperature, E is the activation energy of the rate-determining step in the chemical reaction, and k = 8.6×10−5 eV/K is the Boltzmann constant. In Hallberg and Peck, the authors show that this relationship applies well to measurements of mean time to failure for temperatures 20-150 °C, relative humidity 20%-100, and activation energy 0.9 eV. Our devices operate at the rodent body temperature of 37°C = 310 K. During our accelerated aging test at 60°C = 333 K, we expect an acceleration of ×10.
[13-JUL-21] In our Technical Proposal, we laid out our plan to develop a transmitter small enough to be implanted in the body of a rat, fast enough to transmit four hundred data samples per second, powerful enough to be detected at a range of three meters, and efficient enough to operate for three months on a lithium battery. We divided our development into four stages.
The electronics of Stage Four were ready in March of 2007. It was then that we confronted the problem of robust, water-proof encapsulation for the transmitters. At the time we wrote our Technical Proposal, we believed that two coats of silicone dispersion would be adequate to protect the transmitter from water. Given the simplicity of the procedure, there would be no point in coating the transmitters ourselves. We proposed to send transmitters to our users without batteries, wires, or coating. Our user could solder their preferred wires to the transmitters, install the battery, and apply the coating.
Archive Photograph: See here for photograph of the prototype electronics for Stage Four in 2007. The receiver is the aluminum box with its lid off. Alice is holding a transmitter (prototype version with external battery and programming connector). The loop antenna on a cable sits at one end of the table. A dipole antenna (disconnected) is next to the loop. Two Modulating Transmitters are in the background. The black box is the LWDAQ Driver, connected to our computer. All circuits shown in the picture are now discontinued, but their replacements look much the same. The LWDAQ Driver is a black box, the receiver remains a silver box.
Water-proof encapsulation is not straightforward. Capillary action makes water a relentless invader of any opening or crack. During encapsulation, air bubbles trapped beneath components emerge into the curing encapsulation. In the low pressure of an aircraft cargo hold, the bubble beneath the battery pushes outwards, and will burst an encapsulation made of silicone. Our work on encapsulation is an appendix to the work we committed to in our Technical Proposal. After nine months of effort, we arrived at an encapsulation process that uses both epoxy and silicone, which we called Process X. We made our first transmitters with Process X in December 2007. We describe our work on encapsulation and encapsulation methods in our Encapsulation report. We later improved Process X and arrived at Process B. We made our first transmitters with Process B in June 2009.
Once our transmitters were water-proof in Process X, they endured for long enough in live animals for the wires to break from repetitive stress. Wires broke at the neck of a rat, at their solder joints, and where they emerged from epoxy encapsulation. Our work on flexible wires is an appendix to the work we committed to in our Technical Proposal, and was funded in part by ION's purchase of ten prototype transmitters. We describe our work on wire fatigue in our Flexible Wires report. In Spring 2009 we began trials using a variety of steel wires, including steel springs, as we describe in the Trials section of Flexible Wires. We eventually arrived at a stranded wire for the antenna and helical steel wires for the input leads. We insulate both with silicone, and this insulation forms part of the outer cover of the transmitter body.
Interference with our subcutaneous transmitter signals from outside sources proved to be a problem in ION's London laboratory. We solved the problem of interference with Faraday enclosures. Our work on Faraday enclosures is an appendix to the work we committed to in our Technical Proposal, and was funded in part by ION's purchase of ten more prototype transmitters. We describe the development of a practical Faraday enclosure, and document its success, in Faraday enclosures. Faraday enclosures are now an integral part of the subcutaneous transmitter system. Not only do they provide immunity to outside interference, but they allow many transmitters to operate simultaneously in the same space without interfering with one another.
At the end of October 2013, we had prototypes of our new Dual-Channel EEG Monitor (A3028). We were working on an automated thermo-plastic inner encapsulation for these devices, followed by silicone dipping. We were building the first of our Octal Data Receivers (A3027).
At the end of January 2017, the Octal Data Receiver (A3027E, ODR) with firmware version ≥12 supports channel numbers 1-222. We can, in principle, operate 196 transmitters in the same Faraday enclosure and record data from all of them with a single A3027E. The Animal Location Tracker (A3032A) allows us to track the movements of animals over a 16 cm × 32 cm platform. Our Implantable Sensor with Lamp (A3030D) is receiving commands and flashing its optogenetic stimulator while implanted in rats. We have manufactured over two thousand transmitters to date, and are currently shipping fifty of them per month.
In July 2021 we released the Animal Location Tracker (A3038A, ALT), designed to measure activity and record telemetry signals simultanously. The ALT is our first receiver with Power over Ethernet, and as a result greatly simplifies the set-up of the telemetry system. To support recording from eight ALTs at a time, we split the Neuroarchiver Tool into two parts, Neuroplayer and Neurorecorder, that run in independent recording and playback processes. We no longer have to worry about recording and playback interfering with one another, nor one recording affecting another. In March 2022 we released the Head-Mounting Transmitter (A3040, HMT), which mounts by connector to the skull of an animal, and has a battery we can replace.
In October 2022 we began shipping the Telemetry Control Box (TCB-A16). It provides sixteen independent coaxial antenna inputs, compared to the ODR's eight inputs, and in addition provide antenna power measurements that will allow activity monitoring within cage racks and location tracking in large habitats. Our proposed TCB-B16 will add command transmitters to the controller so that we can operate Implantable Stimulator-Transponders (A3041) and other implantable devices equipped with crystal radio command receivers.
