Name: Design and Construction of a Cost Effective Headstage for Simultaneous Neural Stimulation and Recording in the Water Maze (Video) | JoVE Journal
Uploaded: 2010-10-13T17:00:00.000+00:00
Duration: 9 min 9 s
Description: Learn to build a cost-effective headstage amplifier for simultaneous neural recording and stimulation in awake behaving animals. This customizable method enables reliable local field potential recordings from the rat hippocampus.

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We would like to acknowledge Justin Riceberg for technical assistance in the testing of the headstage. This work was supported in part by National Institutes of Health Grants MH65658, MH073689, and F30 AG034003-01A1, and the Mount Sinai School of Medicine.

1. Headstage Design and Fabrication Overview
A completed headstage pre-amplifier system consists of the following components:
<ol>
	<li>A computer designed integrated circuit board with four channels of recording and two channels of stimulation.</li>
	<li>Electrodes that relay the recordings and stimulation channels to the appropriate connectors on the animals head stage.</li>
	<li>A tether cable and interface that relays the recording and stimulation channels to a slip-ring commutator.</li>
	<li>Two forms of waterproofing on the head stage and on the integrated circuit.</li>
</ol>
2. Design and Layout of the Headstage
We designed a printed integrated circuit board with 4 source followers and 2 pass through stimulation channels using a commercially available software CAD package (www.expresspcb.com). One headstage is designed, and the design pattern is tiled across a larger sheet of circuit board to provide many headstages at a lower cost. The CAD file is transmitted to the printed circuit board (PCB) manufacturing service where double sided, two layer boards are printed using top and bottom copper layers with all holes plated through. The boards use tin/lead solder plated traces and pads that correspond to industry standard FR-4 laminate. The PCB can be ordered and shipped the next business day. Individual headstages are cut from the large tiled PCB sheet and electronic components are connected with solder. An example layout of the headstage used in this protocol is shown (Figure 1). Please see the manufacturer website for detailed instructions and tips on designing a PCB.
3. Preparation and Soldering of Electronic Components
All electronic components used in this protocol are of the Surface Mount Device (SMD) type. The footprint of each device type (resistor, capacitor, etc.) was chosen to minimize wasted space on the headstage, but larger SMD components may be used if necessary.
<ol>
	<li>Organize your components on the circuit board and prepare them for soldering.</li>
	<li>Place a small amount of solder on the tin/lead pad to be soldered and bring the tip of the soldering iron close to melt and adhere the solder to the pad.</li>
	<li>Solder the capacitors first, resistors next, and then the operational amplifiers.</li>
	<li>The op-amps can be destroyed by over heating. We used a variable heat soldering gun and the lowest heat necessary to melt the solder. Minimize the heating time used to make a good solder joint. We found it convenient to create a small reservoir of solder that flows into the footprint pad by applying solder along the communication or power traces.</li>
	<li>Test that the resistance across your resistors meets the expected value using an ohmmeter.</li>
	<li>Inspect each pad closely to ensure that no solder is connecting or bridging adjacent pads. If there is a solder bridge, heat and melt the solder and use a tweezer to break the bridge.</li>
</ol>
4. Assembly and Fabrication of Implant and Tether Interfaces
<ol>
	<li>Insert a 23 Ga hypodermic tubing into one of the unused ports of the 9pin ABS socket for mechanical support.</li>
	<li>Cut six 1 cm length pieces of insulated magnetic wire for connecting the female amphenol pins to the ABS socket.</li>
	<li>Solder the female pins to the magnetic wire using a small amount of solder in each pin, and by heating the pin from the tip.</li>
	<li>Solder the other end of each magnetic wire into the ABS sockets.</li>
	<li>Position the male - female MillMax pins over the pads on the tether side of the headstage, and squeeze the pins over the PCB to ensure a pressure fit. Then, solder each pin to each pad in order.</li>
	<li>Position the ring nut over the neural implant connector and push all the female pins into the desired slots on the plastic base.</li>
	<li>Apply acrylic glue (krazy glue) to the top of the ABS socket connector and secure it to the base of the headstage. Glue the 23Ga tube to the tether side of the headstage for support.</li>
</ol>
5. Waterproofing the Headstage
<ol>
	<li>To waterproof the headstage, encase the entire integrated circuit and any exposed metal surfaces with 5 minute epoxy. Allow 5-10 minutes for initial drying, and 60 minutes for permanent drying.</li>
	<li>Poster Tack putty should be applied to the surface the 9 pin ABS socket around the female pins protruding from the animal end.</li>
	<li>For maximal waterproofing, the ABS socket and ABS plug should be mated and the ring nut should be tightened. A small strip of parafilm should be wrapped around the interface to prevent water from contacting the pin connectors.</li>
</ol>
Secrets to Success
<ol>
	<li>When soldering the operational amplifier, do not overheat the plastic package. If there is visible burning or melting, remove the op-amp and replace it. Applying too much heat may damage the op amp without visible melting. Test each channel with a function generator after soldering.</li>
	<li>Make sure there are no stray connections or solder bridges on the headstage. Solder wick can be used to remove components or stray solder.</li></ol>
6. Representative Results
A good recording will follow an input source signal without cutting in and out during movement. To test the headstage, the user should use a function generator as an input to each recording pin and evaluate the waveforms at the Mill Max header output end. Physically moving the headstage should not influence the shape of recorded waveforms.
<img src="/files/ftp_upload/2155/2155fig1.jpg" alt="PCB design diagrams; A and B circuits with traces for electronic signal pathways analysis." data-altupdatedat="1788235205000" data-alt="Figure 1"> Figure 1. (A) Schematic view of top layer of printed circuit board (PCB). Red lines indicate copper traces. Yellow lines indicate optional silkscreen layer for positioning of electronic parts. Shown are the operational amplifier placement (central yellow square), resistors (smaller yellow squares) and capacitors (smallest yellow squares). (B) Schematic view of bottom layer of PCB. Green lines indicate copper traces. Large green filled area represents ground plate. A model file of this schematic is provided for download.

<ol><li>Morris, R. G., Garrud, P., Rawlins, J. N., O'Keefe, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Place+navigation+impaired+in+rats+with+hippocampal+lesions%5BTitle%5D)+AND+%22Nature%22%5BJournal%5D)">Place navigation impaired in rats with hippocampal lesions.</a> Nature. 297, 681-683 (1982).</li>
<li>Rolston, J. D., Gross, R. E., Potter, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+low-cost+multielectrode+system+for+data+acquisition+enabling+real-time+closed-loop+processing+with+rapid+recovery+from+stimulation+artifacts.+Front%5BTitle%5D)+AND+%22Neuroengineering%22%5BJournal%5D)">A low-cost multielectrode system for data acquisition enabling real-time closed-loop processing with rapid recovery from stimulation artifacts. Front.</a> Neuroengineering. 2, 12-12 (2009).</li>
<li>Fee, M. S., Leonardo, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Miniature+motorized+microdrive+and+commutator+system+for+chronic+neural+recording+in+small+animals%5BTitle%5D)+AND+%22J.+Neurosci.+Methods%22%5BJournal%5D)">Miniature motorized microdrive and commutator system for chronic neural recording in small animals.</a> J. Neurosci. Methods. 112, 83-94 (2001).</li>
<li>Shirvalkar, P. R., Rapp, P. R., Shapiro, M. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Bidirectional+changes+to+hippocampal+theta+gamma+oscillations+predict+memory+for+recent+spatial+episodes%5BTitle%5D)+AND+%22Proc+Natl+Acad+Sci+U+S+A%22%5BJournal%5D)">Bidirectional changes to hippocampal theta gamma oscillations predict memory for recent spatial episodes.</a> Proc Natl Acad Sci U S A. 107 (15), 7054-709 (2010).</li>
</ol>

No conflicts of interest declared.

Water maze based behavioral tasks provide effective tools to probe animals' cognitive abilities and representations in real time 1. The use of lightweight and cost effective headstage preamplifiers is especially critical in behavioral neurophysiology applications involving the water maze. Because escaping the cold water provides intrinsic motivation for animals in these tasks, their behavior can be tested in repeated trials without the difficulties of motivating animals on dry mazes. The low cost headstage pre...

<table><tbody><tr><td>Custom designed Integrated Circuit Board</td><td>ExpressPCB.com</td><td>Custom</td><td>2 layer double sided PCB</td></tr><tr><td>Surface mount Resistors</td><td>Mouser Electronics</td><td>RK73B1JTTDD156J</td><td>1/10 W, 15 M Ohm 0603, Thick film</td></tr><tr><td>SMD Capacitors </td><td>Newark Inc</td><td>0402YD104KAT2A</td><td>0.1 uF (0402)</td></tr><tr><td>SMD Operational Amplifier (quad FET)</td><td>Mouser Electronics</td><td>595-TLC2274AIPW</td><td>Quad FET - low noise</td></tr><tr><td>9 pin ABS socket</td><td>Ginder Scientific</td><td>GS09SKT-220</td><td>For animal connection</td></tr><tr><td>9-pin ABS Plug</td><td>Ginder Scientific</td><td>GS09PLG-220</td><td>To be implanted in animal</td></tr><tr><td>Black Delrin Ring Nut</td><td>Ginder Scientific</td><td>GS09BDN-220</td><td></td></tr><tr><td>Black Delrin Cap Nut</td><td>Ginder Scientific</td><td>GS09BDC-220</td><td>For protecting animal&amp;rsquo;s implant</td></tr><tr><td>Amphenol pins (male)</td><td>Ginder Scientific</td><td>220-P02</td><td></td></tr><tr><td>Amphenol pins (female)</td><td>Ginder Scientific</td><td>220-S02</td><td></td></tr><tr><td>5-minute epoxy</td><td>Devcon Inc.</td><td>S-208 / 20845</td><td></td></tr><tr><td>Poster Tack Putty</td><td>Elmer&amp;rsquo;s</td><td>026000015318</td><td></td></tr><tr><td>Rosin Core Solder</td><td>Radio Shack</td><td>64-009</td><td></td></tr><tr><td>Soldering Iron </td><td>Weller</td><td>Pyropen WSTA6</td><td>(adjustable heat)</td></tr><tr><td>Metal tweezers</td><td>Fine Science Tools</td><td>Size 5</td><td></td></tr><tr><td>Parafilm sheets</td><td>American National Can</td><td>Parafilm 4&quot; x 250&amp;rsquo;</td><td></td></tr><tr><td>Magnet wire</td><td>Belden</td><td>8052</td><td>Enamel Coated</td></tr><tr><td>Ammeter/ Multimeter</td><td>Fluke Biomedical</td><td>77 Multimeter</td><td></td></tr><tr><td>Millmax Double Row Headers</td><td>Mill-Max</td><td>575 - 002101</td><td>0.05&quot;</td></tr><tr><td>Solder Wick</td><td>Chemtronics</td><td>7-10L</td><td>0.75 in.</td></tr></tbody></table>

design and construction of a cost effective headstage for simultaneous neural stimulation and recording in the water maze

Design and Construction of a Cost Effective Headstage for Simultaneous Neural Stimulation and Recording in the Water Maze

Headstage preamplifiers and source followers are commonly used to study neural activity in behavioral neurophysiology experiments. Available commercial products are often expensive, not easily customized, and not submersible. Here we describe a method to design and build a customized, integrated circuit headstage for simultaneous 4-channel neural recording and 2-channel simulation in awake, behaving animals. The headstage is designed using a free, commercially available CAD-type design package, and can be modified easily to accommodate different scales (e.g. to add channels). A customized printed circuit board is built using surface mount resistors, capacitors and operational amplifiers to construct the unity gain source follower circuit. The headstage is made water-proof with a combination of epoxy, parafilm and a synthetic rubber putty. We have successfully used this device to record local field potentials and stimulate different brain regions simultaneously via independent channels in rats swimming in a water maze. The total cost is &lt; $30/unit and can be manufactured readily.

We present a low-cost method to design and construct a light headstage pre-amplifier system with simultaneous neural recording and stimulation capability. This device can be waterproofed for use in swimming animals.

Headstage preamplifiers and source followers are commonly used to study neural activity in behavioral neurophysiology experiments. Available commercial ...

This video demonstrates a procedure to construct a head stage pre amplifier system, which simultaneously records and stimulates chronically and implanted electrodes in awake behaving animals. First, a printed circuit board is designed for a head stage with a targeted number of recording and stimulating channels. Two examples of source follower circuits are shown.The electronic components are then sold to the circuit board and the head stage and tether cable connectors are constructed to provide an interface between the head stage and the animal's implant. With no loose connections in the system, reliable local field potential recordings can be obtained from the rat hippocampus. The main advantage of this technique over existing methods like purchasing a commercially available head stage amplifier system is that this method can be thousands of dollars cheaper and easily customized for specific stimulation and Recording experiments.Because this method allows simultaneous recording and stimulation of the behaving brain, it can help identify and control neural mechanisms of cognition. In turn, these methods may ultimately help to improve function and treat brain disorders in people. The low cost head stage pre amplifier system featured in this video allows simultaneous neural recording and stimulation and can be made waterproof for use with swimming animals.A completed system consists of the following components, a computer designed integrated circuit board with four recording channels and two stimulation channels. Gold pins relay brain signals to the head stage and stimulation signals to the brain by a separate connectors on the animal's head stage. A tether cable and connector relays the recording and stimulation channels through a slip ring commutator to a computerized physiology workstation.There are two forms of waterproofing on the head stage and on the integrated circuit. Now let's take a look at the design and layout of the head stage using a commercially available software CAD package. A printed integrated circuit board with four unity gain source followers and two pass through stimulation channels is designed First, create connector pads on both sides for the pin connectors on the tether and animal connection.A quad FET operational amplifier can accept up to four channels of plus minus input. Two power lines are needed, each of which can be connected to ground fire. 0.1 micro ferre capacitors to prevent possible induced oscillations in the operational amplifiers.To make a unity gain amplifier circuit, route the signal into the positive input of the operational amplifier and then connect the output back to the inverting input. Then add 10 mega ohm resistors in series with the positive input to ground for the stimulation channels. Lay pass through traces connecting the outputs to inputs using VS.To guide the traces to the other layer of the bored andm back.Finally, create a large ground plate on the backside layer to connect all vias that are meant the ground. The ground plate then should be connected to the ground pin, out to the tether and animal side. A single head stage is designed and the design pattern is tiled across a larger sheet of circuit board to provide many head stages at a lower cost.The CAD file is transmitted to the printed circuit board manufacturing service where double-sided two layer boards are printed using top and bottom copper layers with all holes plated through the boards. Use tin lead, solar plated traces and pads that correspond to industry standard FR four laminates. Individual head stages are cut from the large tiled PCB sheet and electronic components are connected with solar.Now let's see how to connect the electronic components. Begin preparations for soldiering of electronic components by organizing the components on the circuit board to prepare them for soldiering. Place a small amount of sold on the tin lead pad to be soldered and bring the tip of the soldiering iron close to melt in a deer.The soldier to the pad soldier the capacitors first resistors next, and then the operational amplifiers. Because the op amps can be destroyed by overheating is a variable heat soldiering gun, and the lowest heat necessary to melt the soldier. It is best to minimize the heating time used to make a good soldier joint.We find it convenient to create a small reservoir or soldier that flows into the footprint pad by applying soldier along the communication or power traces. Now use an O meter to test that the resistance across the resistors meets the expected value. Inspect each pad closely to ensure that no soldier is connecting or bridging adjacent pads.If there is a soldier bridge here to melt the soldier and use tweezers to break the bridge. Now let's see how to assemble and fabricate tether interfaces. First, insert a piece of 19 gauge hypodermic tubing into one of the unused ports of the nine pin A BS socket for mechanical support.Cut six two centimeter length pieces of insulated magnetic wire for connecting the female and phenol pins to the A BS socket. Now using a small amount of shoulder in each pin, heat the pin from the tip to shoulder the female pins to the magnetic wire. Position the male female milax pins over the pads on the tether side of the head stage and squeeze the pins over the PCB to ensure a pressure fit.Then sold each pin to each pad in order. Next, position the ring nut over the neural implant connector and push all the female pins into the desired slots on the plastic base. Apply acrylic glue to the top of the A BS socket connector and secure it to the base of the head stage.Finally, glue the 19 gauge tube to the tether side of the head stage for support. Now let's see how to waterproof the head stage to waterproof the head stage in case the entire integrated circuit and any exposed metal surfaces with five minutes epoxy. Allow five to 10 minutes for initial curing and 60 minutes for permanent curing.Apply post attack putty to the surface of the nine pin a BS socket around the female pins protruding from the animal end. For maximal waterproofing, the A BS socket and a BS plug should be mated and the ring nut should be tightened. Wrap a small strip of para film around the interface to prevent water from contacting the pin connectors.Now we'll show some representative results shown here. As an example of a successful recording. A good recording will follow an input source signal without cutting in and out during movement.For a good reference on electrophysiology or LFPS refer to citation two. When passing a current through a particular input channel, the user should ensure that there is no crosstalk of the signal into adjacent channels. This can be done by passing the signal through each channel and testing the output of all other channels.If there is crosstalk between channels, please refer to reference article three for useful suggestions. The user may desire the head stage to be positioned flat on the animal's head or with additional channels. We have successfully manufactured such head stages with additional channels and configurations Once mastered.This technique can be done in an assembly line fashion to build multiple head stages at the same time so we can establish replicability and provide a quick backup for Damaged head stages. This procedure can be used in combination with other methods such as targeted drug infusion or optogenetic stimulation to investigate how different neurotransmitters and neuromodulators contribute to brain mechanisms of memory. And we used this combination of techniques to record and modify brain rhythms in the rat hippocampus, which predicted memory performance.

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Learn to build a cost-effective headstage amplifier for simultaneous neural recording and stimulation in awake behaving animals. This customizable method enables reliable local field potential recordings from the rat hippocampus.

Design and Construction of a Cost Effective Headstage for Simultaneous Neural Stimulation and Recording in the Water Maze

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