Name: autonomous and rechargeable microneurostimulator endoscopically implantable into the submucosa
Uploaded: 2018-09-27T12:30:00
Description: Watch this Scientific Journal Video about Autonomous and Rechargeable Microneurostimulator Endoscopically Implantable into the Submucosa at JoVE.com

The authors declare that they have no competing financial interests.

All endoscopic procedures including animal subjects have been approved at the Institute of Animal Physiology and Genetics, Academy of Science Czech Republic (Biomedical Center PIGMOD), Libechov, Czech Republic (project Experiments in implantation of battery-less and battery devices into submucosa of the esophagus and stomach &#8212; experimental study). All experiments are done in compliance with Czech law 246/1992 Sb. &#34;On the protection of animals against maltreatment, as amended&#34;.&#160;Transmitter device is not...

<ol>
	<li>Abell, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gastric+electrical+stimulation+for+medically+refractory+gastroparesis.">Gastric electrical stimulation for medically refractory gastroparesis.</a> Gastroenterology. 125 (2), 421-428 (2003).</li><li>O'Grady, G., Egbuji, J., Du, P., Cheng, L. K., Pullan, A. J., Windsor, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High-frequency+gastric+electrical+stimulation+for+the+treatment+of+gastroparesis:+a+meta-analysis.">High-frequency gastric electrical stimulation for the treatment of gastroparesis: a meta-analysis.</a> World J Surg. 33 (8), 1693-1701 (2009).</li><li>Chu, H., Lin, Y., Zhong, L., McCallum, R. W., Hou, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Treatment+of+high-frequency+gastric+electrical+stimulation+for+gastroparesis.">Treatment of high-frequency gastric electrical stimulation for gastroparesis.</a> J Gastroenterol Hepatol. 27 (6), 1017-1026 (2012).</li><li>Rodr&#237;guez, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrical+stimulation+therapy+of+the+lower+esophageal+sphincter+is+successful+in+treating+GERD:+final+results+of+open-label+prospective+trial.">Electrical stimulation therapy of the lower esophageal sphincter is successful in treating GERD: final results of open-label prospective trial.</a> Surg Endosc. 27 (4), 1083-1092 (2013).</li><li>Ellis, F., Berne, T. V., Settevig, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+prevention+of+experimentally+induced+reflux+by+electrical+stimulation+of+the+distal+esophagus.">The prevention of experimentally induced reflux by electrical stimulation of the distal esophagus.</a> Am J Surg. 115, 482-487 (1968).</li><li>Rinsma, N. F., Bouvy, N. D., Masclee, A. A. M., Conchillo, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrical+Stimulation+Therapy+for+Gastroesophageal+Reflux+Disease.">Electrical Stimulation Therapy for Gastroesophageal Reflux Disease.</a> J Neurogastroenterol. 20 (3), 287-293 (2014).</li><li>Medtronic Inc, . <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Enterra Therapy 3116 - Gastric Electrical Stimulation System. , (2016).</li><li>Rodriguez, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Two-year+results+of+intermittent+electrical+stimulation+of+the+lower+esophageal+sphincter+treatment+of+gastroesophageal+reflux+disease.">Two-year results of intermittent electrical stimulation of the lower esophageal sphincter treatment of gastroesophageal reflux disease.</a> Surgery. 157 (3), 556-567 (2015).</li><li>Hajer, J., Nov&#225;k, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+an+Autonomous+Endoscopically+Implantable+Submucosal+Microdevice+Capable+of+Neurostimulation+in+the+Gastrointestinal+Tract.">Development of an Autonomous Endoscopically Implantable Submucosal Microdevice Capable of Neurostimulation in the Gastrointestinal Tract.</a> Gastroent Res Pract. , 8098067 (2017).</li><li>Deb, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+innovative+techniques+for+the+endoscopic+implantation+and+securing+of+a+novel,+wireless,+miniature+gastrostimulator+(with+videos).">Development of innovative techniques for the endoscopic implantation and securing of a novel, wireless, miniature gastrostimulator (with videos).</a> Gastrointest. Endosc. 76 (1), 179-184 (2012).</li><li>Jiang, G., Zhou, D. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Technology advances and challenges in hermetic packaging for implantable medical devices. , (2017).</li><li>Vonthein, R., Heimerl, T., Schwandner, T., Ziegler, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrical+stimulation+and+biofeedback+for+the+treatment+of+fecal+incontinence:+a+systematic+review.">Electrical stimulation and biofeedback for the treatment of fecal incontinence: a systematic review.</a> Int J Colorectal Dis. 28 (11), 1567-1577 (2013).</li></ol>

The design of the implantable device should primarily focus on the overall size of the device, achievable stimulation profiles (maximum voltage, maximum deliverable current, length of pulses and pulse frequency). Main limitation from the hardware perspective is the size and availability of suitable components. To minimize the overall size, surface mount components are preferred because of their compact packaging. The best solution would be to integrate bare chip dies on the substrate. However, this is limited by both the...

Gastric dysmotility can be a sign of several relatively common diseases such as gastroparesis, which is usually characterized by a chronic progression and imposes rather severe consequences on the social, work-related, and physical status of the patient. Most cases of gastroparesis are usually diabetic or idiopathic in origin and are often resistant to available medication1. Patients afflicted with this condition most commonly present with nausea and repeated vomiting. Based on previous research, it is known that the application of high-frequency low-energetic electrical stimulation can help to effectively moderate and alleviate the symptoms of gastric dysmotility1,2.
Based on previous studies, it is proven that high-frequency gastric electrical stimulation can significantly improve the symptoms and gastric emptying3. It has also been shown that lower esophageal sphincter neurostimulator therapy is safe and effective for the treatment of gastroesophageal reflux disease (GERD), reducing the acid exposure and eliminating daily proton-pump inhibitor (PPI) usage without stimulation related adverse effects4. Before human trials, first studies were performed in animal models (canine models5). Based on these studies, electrical stimulation of the lower esophageal sphincter (LES, 20 Hz, pulse width of 3 ms) caused a prolonged contraction of the LES5. Similar effects of high (20 Hz, pulse width of 200 &#956;s) and low (6 cycles/min, pulse width of 375 ms) frequency electrical stimulation on LES in GERD patients were investigated. Both high and low frequency stimulation were effective6. However, currently, there are only two neurostimulation devices for gastric or esophageal stimulation available on the market7,8. In those devices, the electrodes can be implanted surgically, laparoscopically or robotically. The device itself is implanted subcutaneously. This requires general anaesthesia and have a bulky device fitted, using intramuscular catheters which allow for the stimulation of the gastric or esophageal muscle tissue. So, the option of using a wirelessly communicating device implanted into the gastric submucosal layer would represent a definite advantage and improvement in patient comfort. As stated in the previous research9,10, it was proven that an implantation of a miniature neurostimulator into submucosa is possible. For the endoscopic submucosal implantation, we use a technique called endoscopic submucosal pocketing (ESP), based on endoscopic submucosal tunnel dissection10. The goal of this research is to further improve this concept of an implantable neurostimulator, primarily in the scope of power management (specifically the wireless recharging capability), conformity with respective laws and regulations for wireless communication links in medical implantable devices and possibility of bipolar neurostimulation. Next, the presented microneurostimulator is capable of bidirectional communication and the stimulation parameters can be changed in real-time, even while the device is implanted.
This technique is suitable for teams with a therapeutic endoscopist experienced in endoscopic pocketing or tunnel dissections. Next, a hardware and embedded software designer with experience in building hardware prototypes with microcontrollers and radio frequency circuits using surface mount technology is needed. For building the hardware prototypes, a lab equipped with a reflow soldering station and basic equipment for electrical measurements (at least a digital multimeter, an oscilloscope, a spectrum analyzer and PICkit3 programmer) is required.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>EIA 0402 ceramic capacitor 1.8 pF</td>
			<td>AVX</td>
			<td>04025U1R8BAT2A</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>EIA 0402 ceramic capacitor 100 nF</td>
			<td>TDK</td>
			<td>CGA2B3X7R1H104K050BB</td>
			<td>7 pcs</td>
		</tr>
		<tr>
			<td>EIA 0402 ceramic capacitor 100 pF</td>
			<td>Murata Electronics</td>
			<td>GRM1555C1H101JA01D</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>EIA 0402 thick film resistor 10 k&#937;</td>
			<td>Vishay</td>
			<td>CRCW040210K7FKED</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>EIA 0402 ceramic capacitor 10 nF</td>
			<td>Murata Electronics</td>
			<td>GRM155R71C103KA01D</td>
			<td>3 pcs</td>
		</tr>
		<tr>
			<td>EIA 0402 ceramic capacitor 10 pF</td>
			<td>Murata Electronics</td>
			<td>GJM1555C1H100JB01D</td>
			<td>3 pc</td>
		</tr>
		<tr>
			<td>EIA 0402 ceramic capacitor 12 pF</td>
			<td>Murata Electronics</td>
			<td>GJM1555C1H120JB01D</td>
			<td>2 pcs</td>
		</tr>
		<tr>
			<td>EIA 0402 ceramic capacitor 18 pF</td>
			<td>KEMET</td>
			<td>C0402C180J3GACAUTO</td>
			<td>2 pcs</td>
		</tr>
		<tr>
			<td>EIA 0402 resistor 1 m&#937;</td>
			<td>Vishay</td>
			<td>MCS04020C1004FE000</td>
			<td>2 pcs</td>
		</tr>
		<tr>
			<td>EIA 0402 resistor 1 k&#937;</td>
			<td>Yageo</td>
			<td>RC0402FR-071KL</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>EIA 0402 ceramic capacitor 1 nF</td>
			<td>Murata Electronics</td>
			<td>GRM1555C1H102JA01D</td>
			<td>3 pcs</td>
		</tr>
		<tr>
			<td>EIA 0603 ceramic capacitor 2.2 uF</td>
			<td>Murata Electronics</td>
			<td>GCM188R70J225KE22D</td>
			<td>2 pcs</td>
		</tr>
		<tr>
			<td>EIA 0402 resistor 220 k&#937;</td>
			<td>Vishay</td>
			<td>CRCW0402220KJNED</td>
			<td>5 pcs</td>
		</tr>
		<tr>
			<td>0805 22 uH inductor</td>
			<td>TDK</td>
			<td>MLZ2012N220LT000</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>EIA 0402 resistor 330 k&#937;</td>
			<td>Vishay</td>
			<td>CRCW0402330KFKED</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>EIA 0603 ceramic capacitor 4.7 uF</td>
			<td>TDK</td>
			<td>C1608X6S1C475K080AC</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>EIA 0402 resistor 470 &#937;</td>
			<td>Vishay</td>
			<td>RCG0402470RJNED</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>EIA 0402 resistor 470 k&#937;</td>
			<td>Vishay</td>
			<td>CRCW0402470KJNED</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>EIA 0603 inductor 470 nH</td>
			<td>Murata Electronics</td>
			<td>LQW18ANR47G00D</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>EIA 0402 resistor 47 k&#937;</td>
			<td>Murata Electronics</td>
			<td>CRCW040247K0JNED</td>
			<td>2 pcs</td>
		</tr>
		<tr>
			<td>27.0000 MHz crystal 5032</td>
			<td>AVX / Kyocera</td>
			<td>KC5032A27.0000CMGE00</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>EIA 0402 capacitor 6.8 pF</td>
			<td>Murata Electronics</td>
			<td>GJM1555C1H6R8CB01D</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>EIA 0402 inductor 82 nH</td>
			<td>EPCOS / TDK</td>
			<td>B82498F3471J</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>ABS05 32.768 kHz crystal</td>
			<td>ABRACON</td>
			<td>ABS05-32.768KHZ-T</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>CDBU00340-HF schottky diode</td>
			<td>COMCHIP technology</td>
			<td>CDBU00340-HF</td>
			<td>2 pcs</td>
		</tr>
		<tr>
			<td>CG-320S Li-Ion pinpoint battery</td>
			<td>Panasonic</td>
			<td>CG-320S</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>HSMS282P schottky diode rectifier</td>
			<td>Broadcom / Avago</td>
			<td>HSMS-282P-TR1G</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>MAX8570 step-up converter</td>
			<td>Maxim Integrated</td>
			<td>MAX8570EUT+T</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>MICRF113 RF transmitter</td>
			<td>Microchip Technology</td>
			<td>MICRF113YM6-TR</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>4.3 V Zener diode</td>
			<td>ON Semiconductor</td>
			<td>MM3Z4V3ST1G</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>OPA237 operational amplifier</td>
			<td>Texas Instruments</td>
			<td>OPA237N</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>PIC16LF1783 8-bit microcontroller</td>
			<td>Microchip Technology</td>
			<td>PIC16LF1783-I/ML</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>TPS70628 low-drop regulator</td>
			<td>Texas Instruments</td>
			<td>TPS70628DBVT</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>EIA 1206 thick film resistor 0 &#937;</td>
			<td>Yageo</td>
			<td>RC1206JR-070RL</td>
			<td>2 pcs</td>
		</tr>
		<tr>
			<td>EIA 0603 thick film resistor 0 &#937;</td>
			<td>Yageo</td>
			<td>RC0603JR-070RL</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>EIA 0402 thick film resistor 100 k&#937;</td>
			<td>Yageo</td>
			<td>RC0402FR-07100KL</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>EIA 0603 thick film resistor 100 k&#937;</td>
			<td>Yageo</td>
			<td>RC0603FR-07100KL</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>EIA 0805 ceramic capacitor 100 nF</td>
			<td>KEMET</td>
			<td>C0805C104K5RAC7210</td>
			<td>2 pcs</td>
		</tr>
		<tr>
			<td>EIA 0402 thick film resistor 10 k&#937;</td>
			<td>Yageo</td>
			<td>RC0402JR-0710KL</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>EIA 1206 ceramic capacitor 10 nF</td>
			<td>Samsung</td>
			<td>CL31B103KHFSW6E</td>
			<td>2 pcs</td>
		</tr>
		<tr>
			<td>EIA 0402 thick film resistor 1 k&#937;</td>
			<td>Yageo</td>
			<td>RC0402JR-071KL</td>
			<td>2 pcs</td>
		</tr>
		<tr>
			<td>EIA 0402 thick film resistor 220 &#937;</td>
			<td>Yageo</td>
			<td>RC0402JR-07220RL</td>
			<td>2 pcs</td>
		</tr>
		<tr>
			<td>EIA 0402 ceramic capacitor 220 nF</td>
			<td>TDK</td>
			<td>C1005X5R1C224K050BB</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>EIA 1206 ceramic capacitor 22 nF</td>
			<td>TDK</td>
			<td>C3216X7R2J223K130AA</td>
			<td>2 pcs</td>
		</tr>
		<tr>
			<td>SMC B tantalum capacitor 22 uF</td>
			<td>AVX</td>
			<td>TPSB226K010T0700&#160;</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>EIA 0402 thick film resistor 27 &#937;</td>
			<td>Yageo</td>
			<td>RC0402FR-0727RL</td>
			<td>2 pcs</td>
		</tr>
		<tr>
			<td>EIA 1206 thick film resistor 3.3 &#937;</td>
			<td>Yageo</td>
			<td>RC1206JR-073K3L</td>
			<td>3 pcs</td>
		</tr>
		<tr>
			<td>SOT23 3.3V zener diode</td>
			<td>ON Semiconductor</td>
			<td>BZX84C3V3LT1G</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>SMC A tantalum capacitor 4.7uF</td>
			<td>KEMET</td>
			<td>T491A475M016AT</td>
			<td>2 pcs</td>
		</tr>
		<tr>
			<td>EIA 0603 thick film resistor 470 &#937;</td>
			<td>Yageo</td>
			<td>RC0603JR-07470RL</td>
			<td>2 pcs</td>
		</tr>
		<tr>
			<td>EIA 1206 ceramic capacitor 470 nF</td>
			<td>KEMET</td>
			<td>C1206C471J5GACTU</td>
			<td>3 pcs</td>
		</tr>
		<tr>
			<td>Electrolytic capacitor 470 uF</td>
			<td>Panasonic</td>
			<td>EEE-1CA471UP</td>
			<td>3 pcs</td>
		</tr>
		<tr>
			<td>EIA 0402 ceramic capacitor 47 pF</td>
			<td>AVX</td>
			<td>04025A470JAT2A</td>
			<td>2 pcs</td>
		</tr>
		<tr>
			<td>0603 GREEN LED</td>
			<td>Lite-On Inc.</td>
			<td>LTST-C191KGKT</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>0603 RED LED</td>
			<td>Lite-On Inc.</td>
			<td>LTST-C191KRKT</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>16 MHz CX3225 crystal</td>
			<td>EPSON</td>
			<td>FA-238 16.0000MB-C3</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>0805 ferrite bead</td>
			<td>Wurth Electronics Inc.</td>
			<td>742792040</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>IR2110SO FET driver</td>
			<td>Infineon Technologies</td>
			<td>IR2110SPBF</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>FT230XS USB to seri&#225;l converter</td>
			<td>FTDI Ltd.</td>
			<td>FT230XS-R</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>Mini USB connector</td>
			<td>EDAC Inc.</td>
			<td>690-005-299-043</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>PIC16F1783 8-bit microcontroller</td>
			<td>Microchip Technology</td>
			<td>PIC16F1783-I/ML</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>REG1117 3.3 V regulator SOT223</td>
			<td>Texas Instruments</td>
			<td>REG1117-3.3/2K5</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>Schottky SMB diode rectifier</td>
			<td>STMicroelectronics</td>
			<td>STPS3H100UF</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>SMB package TVS diode</td>
			<td>Littelfuse Inc.</td>
			<td>1KSMBJ6V8</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>IRLZ44NPBF N-channel MOSFET</td>
			<td>Infineon Technologies</td>
			<td>IRLZ44NPBF</td>
			<td>2 pcs</td>
		</tr>
		<tr>
			<td>RTL2832U receiver dongle</td>
			<td>EVOLVEO</td>
			<td>Mars</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>PICkit 3</td>
			<td>Microchip Technology</td>
			<td>PICkit 3</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>Mini USB to USB A cable</td>
			<td>OEM</td>
			<td>Mini USB to USB-A</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>Printed circuit board, implantable device</td>
			<td>---</td>
			<td>Manufacture with the provided supplementary file</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>Printed circuit board, transmitter/receiver device</td>
			<td>---</td>
			<td>Manufacture with the provided supplementary file</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>Printed circuit board, implantable device</td>
			<td>---</td>
			<td>Manufacture with the provided supplementary file</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>AWG18 wire</td>
			<td>Alpha Wire</td>
			<td>3055 BK001</td>
			<td>2 m</td>
		</tr>
		<tr>
			<td>AWG42 wire</td>
			<td>Daburn Electronics</td>
			<td>2420/42 BK-100</td>
			<td>1 m</td>
		</tr>
		<tr>
			<td>Olympus GIFQ-160</td>
			<td>Olympus</td>
			<td>N/A (part is obsoleted)</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>Single-use electrosurgical knife with knob-shaped tip and integrated jet function</td>
			<td>Olympus</td>
			<td>KD-655L</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>Single-use oval electrosurgical snare</td>
			<td>Olympus</td>
			<td>SD-210U-15</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>15.5 mm lens hood</td>
			<td>FujiFilm</td>
			<td>DH-28GR</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>Injection therapy needle catheter</td>
			<td>Boston Scientific</td>
			<td>25G</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>Alligator law grasping forceps</td>
			<td>Olympus</td>
			<td>FG-6L-1</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>Instant Mix 5 min epoxy</td>
			<td>Loctite</td>
			<td>N/A</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>Heat shrinkable tubing, inside diameter 9.5 mm</td>
			<td>TE Connectivity</td>
			<td>RNF-100-3/8-X-STK</td>
			<td>1 pc</td>
		</tr>
		<tr>
			<td>ChipQuik solder paste</td>
			<td>Chip Quik</td>
			<td>SMD4300AX10</td>
			<td>1 pc</td>
		</tr>
	</tbody>
</table>

autonomous and rechargeable microneurostimulator endoscopically implantable into the submucosa

Gastric dysmotility can be a sign of common diseases such as longstanding diabetes mellitus. It is known that the application of high-frequency low-energetic stimulation can help to effectively moderate and alleviate the symptoms of gastric dysmotility. The goal of the research was the development of a miniature, endoscopically implantable device to a submucosal pocket. The implantable device is a fully customized electronic package which was specifically designed for the purpose of experiments in the submucosa. The device is equipped with a lithium-ion battery which can be recharged wirelessly by receiving an incident magnetic field from the charging/transmitting coil. The uplink communication is achieved in a MedRadio band at 432 MHz. The device was endoscopically inserted into the submucosal pocket of a live domestic pig used as an in vivo model, specifically in the stomach antrum. The experiment confirmed that the designed device can be implanted into the submucosa and is capable of bidirectional communication. The device can perform bipolar stimulation of muscle tissue.

Figure 17 shows that an endoscopic placement of the gastric neurostimulator into a pocket in submucosa as well as proper placement of the electrodes to the muscular layer was successful. The dimensions of the device (Figure 10) are 35 x 15 x 5 mm3 while the weight is 2.15 g. Figure 17 shows the circuit diagram of the device showing that the device comprises of 6 different modules wh...

Watch this Scientific Journal Video about Autonomous and Rechargeable Microneurostimulator Endoscopically Implantable into the Submucosa at JoVE.com

Autonomous and Rechargeable Microneurostimulator Endoscopically Implantable into the Submucosa

The application of high-frequency low-energetic stimulation can alleviate the symptoms of gastric dysmotility. In this research, a miniature, endoscopically implantable and wirelessly rechargeable device which is implanted into a submucosal pocket is presented. Successful both-way communication and stimulation control were achieved during an experiment on live pig.

Gastric dysmotility can be a sign of common diseases such as longstanding diabetes mellitus. It is known that the application of high-frequency ...

The main advantage of this technique is that it offers a significantly less invasive approach. Generally, individuals new to this method will struggle because the implantable device development is a discipline which requires experience both in hardware and software design. Visual demonstration of this method is critical as the endoscopic submucosal implantation is an experimental procedure currently not used in daily practice.To begin this procedure, place the PCBs on a flat surface. Use a solder paste dispenser with a 0.6 millimeter needle and 60 PSI pressure to manually dispense less than 15 microliters of the soldering paste onto every metallic pad on the PCB. Begin with the top side of the PCB.With a pair of anti-static tweezers, place all components on the top layer of the PCB. Assign the component positions and components to their numbers. After that, use a PCB hot airgun station at 260 degrees Celsius to solder all components.Wait until all the solder paste melts. Put away the hot airgun and allow the board to cool to room temperature. Connect the PCB to all other electrical components except for the battery.Solder the wireless charging communication coil to pads two and three, then connect the antenna to pad one and the PCB electrodes to pads number four and five. Following that, solder the CG-320 battery to pads six and seven. The negative terminal of the battery must be soldered on pad seven.Be careful while performing the next steps. The device is now powered and is sensitive to short circuits and contact with metallic objects. Next, shrink the tubing with a hot airgun heated to 150 degrees Celsius and then allow it to cool.After that, apply epoxy glue to the left end to seal one side of the tubing, then glue the electrode to the back of the PCB with tubing. Subsequently, seal the other end of the tubing. In this step, place the PCBs on a flat surface.Use a solder paste dispenser with 0.6 millimeter needle and 60 PSI pressure to manually dispense less than 50 microliters of the soldering paste onto every metallic pad on the PCB. With a pair of anti-static tweezers, place all components on the top layer of the PCB. Assign the component position and the components to their numbers.Use a PCB hot airgun station preset to 260 degrees Celsius to solder all components. Wait until all the solder paste melts. Put away the hot airgun and allow the board to cool to room temperature.Next, the 12-volt power supply is connected to the wall and the USB connector is inserted to the PC.After that, open the PuTTY software. Select Serial as the connection type, then enter COMX as a serial line where X is the number of the COM Port of the device. If no other COM Port device was installed, this number will be one.Enter 38, 400 as speed. Click Open. The charger transmitter device is now ready to be used.In order to perform the implantation and visualization, insert an animal model dedicated endoscope with the implantable device grasped into a snare into the stomach. When in the stomach, release the device. After that, extract the endoscope.Equip it with a dissection cap and then reinsert it to the stomach. In order to implant the device to submucosa, apply saline solution mixed with methylene blue into the submucosal layer using an injection therapy needle catheter. Make a horizontal incision to create an opening in the submucosa using an electrosurgical knife with a knob-shaped tip.Insert an affixed cap into the newly created space and with the use of an electrosurgical knife, continue disrupting, dilating, and dissecting the submucosal layer to create a large enough pocket to insert the stimulation device. Grasp the device which is lying freely inside the stomach with correctly oriented electrodes with a grasper and navigate it into the submucosal pocket. After that, use two hemostatic clips to secure the device in place inside of the submucosal pocket.Then pull out the endoscope. Put the Ovesco clip on the endoscope and reinsert it into the stomach. Orientate the endoscope towards the pocket, grasp both sides of the pocket and apply the Ovesco.After that, check the correct placement of the Ovesco. After successful implantation, place the charger transmitter coil in proximity of the implanted device, then plug in the RTL2832 dongle in the PC.Run the HDSDR software and set the center frequency to 432 megahertz. Subsequently, select the LO frequency to 431.95 megahertz, then select the Tune frequency to 432.00 megahertz.Select Bandwidth to 960, 000. Transmit a Manchester-coded sequence from the charger transmitter by pressing the R key in the PuTTY terminal and receive the OOK modulated reply from the implant shown in the main HDSDR window. Shown here is the top copper layer of the PCB.The component names are indicated on the top layer. And here is the bottom copper layer of the PCB. The component names are on the bottom layer.And this figure shows the composite picture of all PCB layers. Once mastered, this technique can be done in four hours for the implantable device manufacturing and one hour for device implantation and testing if performed properly. While attempting this procedure, it's important to verify all functions of the implantable device prior to implantation.Following this procedure, other methods like further development of algorithm and data processing can be performed in order to answer additional questions like the efficiency of the method. After its development, this technique paved the way for researchers in the field of gastroenterology to explore third space in gastrointestinal tract. After watching this video, you should have a good understanding of how to perform an endoscopy implantation of autonomous gastric micronuerostimulator in submucosa.Don't forget that working with soldering paste and electronics manufacturing including working with hot airgun can be hazardous and precautions such as gloves and safety glasses should always be worn while performing this procedure.

autonomous-rechargeable-microneurostimulator-endoscopically

Research

JoVE Journal

Neuroscience

Ultrasound-Guided Microinjection into the Mouse Forebrain In Utero at E9.5

In utero survival surgery in mice permits the molecular manipulation of gene expression during development. However, because the uterine wall is opaque during early embryogenesis, the ability to target specific parts of the embryo for microinjection is greatly limited. Fortunately, high-frequency ultrasound imaging permits the generation of images that can be used in real time to guide a microinjection needle into the embryonic region of interest. Here we describe the use of such imaging to guide the injection of retroviral vectors into the ventricular system of the mouse forebrain at embryonic day (E) 9.5. This method uses a laparotomy to permit access to the uterine horns, and a specially designed plate that permits host embryos to be bathed in saline while they are imaged and injected. Successful surgeries often result in most or all of the injected embryos surviving to any subsequent time point of interest (embryonically or postnatally). The principles described here can be used with slight modifications to perform injections into the amnionic fluid of E8.5 embryos (thereby permitting infection along the anterior posterior extent of the neural tube, which has not yet closed), or into the ventricular system of the brain at E10.5/11.5. Furthermore, at mid-neurogenic ages (~E13.5), ultrasound imaging can be used direct injection into specific brain regions for viral infection or cell transplantation. The use of ultrasound imaging to guide in utero injections in mice is a very powerful technique that permits the molecular and cellular manipulation of mouse embryos in ways that would otherwise be exceptionally difficult if not impossible.

Ultrasound-Guided Microinjection into the Mouse Forebrain In Utero at E9.5

In utero survival surgery in mice permits the molecular manipulation of gene expression during development. Here we describe the use of high-frequency ultrasound imaging to guide the injection of retroviral vectors into the mouse brain at embryonic day (E) 9.5.

In utero survival surgery in mice permits the molecular manipulation of gene expression during development.  However, because the uterine wall is opaque ...

Using Insect Electroantennogram Sensors on Autonomous Robots for Olfactory Searches

Robots designed to track chemical leaks in hazardous industrial facilities1 or explosive traces in landmine fields2 face the same problem as insects foraging for food or searching for mates3: the olfactory search is constrained by the physics of turbulent transport4. The concentration landscape of wind borne odors is discontinuous and consists of sporadically located patches. A pre-requisite to olfactory search is that intermittent odor patches are detected. Because of its high speed and sensitivity5-6, the olfactory organ of insects provides a unique opportunity for detection. Insect antennae have been used in the past to detect not only sex pheromones7 but also chemicals that are relevant to humans, e.g., volatile compounds emanating from cancer cells8 or toxic and illicit substances9-11. We describe here a protocol for using insect antennae on autonomous robots and present a proof of concept for tracking odor plumes to their source. The global response of olfactory neurons is recorded in situ in the form of electroantennograms (EAGs). Our experimental design, based on a whole insect preparation, allows stable recordings within a working day. In comparison, EAGs on excised antennae have a lifetime of 2 hr. A custom hardware/software interface was developed between the EAG electrodes and a robot. The measurement system resolves individual odor patches up to 10 Hz, which exceeds the time scale of artificial chemical sensors12. The efficiency of EAG sensors for olfactory searches is further demonstrated in driving the robot toward a source of pheromone. By using identical olfactory stimuli and sensors as in real animals, our robotic platform provides a direct means for testing biological hypotheses about olfactory coding and search strategies13. It may also prove beneficial for detecting other odorants of interests by combining EAGs from different insect species in a bioelectronic nose configuration14 or using nanostructured gas sensors that mimic insect antennae15.

We describe a protocol for using insect antennae in the form of electroantennograms (EAGs) on autonomous robots. Our experimental design allows stable recordings within a day and resolves individual odor patches up to 10 Hz. The efficiency of EAG sensors for olfactory searches is demonstrated in driving a robot toward an odor source.

Robots designed to track chemical leaks in hazardous industrial facilities1 or explosive traces in landmine fields2 face the same problem as insects ...

Stereotaxic Infusion of Oligomeric Amyloid-beta into the Mouse Hippocampus

Alzheimer&#8217;s disease is a neurodegenerative disease affecting the aging population. A key neuropathological feature of the disease is the over-production of amyloid-beta and the deposition of amyloid-beta plaques in brain regions of the afflicted individuals. Throughout the years scientists have generated numerous Alzheimer&#8217;s disease mouse models that attempt to replicate the amyloid-beta pathology. Unfortunately, the mouse models only selectively mimic the disease features. Neuronal death, a prominent effect in the brains of Alzheimer&#8217;s disease patients, is noticeably lacking in these mice. Hence, we and others have employed a method of directly infusing soluble oligomeric species of amyloid-beta - forms of amyloid-beta that have been proven to be most toxic to neurons - stereotaxically into the brain. In this report we utilize male C57BL/6J mice to document this surgical technique of increasing amyloid-beta levels in a select brain region. The infusion target is the dentate gyrus of the hippocampus because this brain structure, along with the basal forebrain that is connected by the cholinergic circuit, represents one of the areas of degeneration in the disease. The results of elevating amyloid-beta in the dentate gyrus via stereotaxic infusion reveal increases in neuron loss in the dentate gyrus within 1 week, while there is a concomitant increase in cell death and cholinergic neuron loss in the vertical limb of the diagonal band of Broca of the basal forebrain. These effects are observed up to 2 weeks. Our data suggests that the current amyloid-beta infusion model provides an alternative mouse model to address region specific neuron death in a short-term basis. The advantage of this model is that amyloid-beta can be elevated in a spatial and temporal manner.

Here, we present a protocol for direct stereotaxic brain infusion of amyloid-beta. This methodology provides an alternative in vivo mouse model to address the short-term effects of amyloid-beta on brain neurons.

Alzheimer&#8217;s disease is a neurodegenerative disease affecting the aging population. A key neuropathological feature of the disease is the ...

Intracerebroventricular and Intravascular Injection of Viral Particles and Fluorescent Microbeads into the Neonatal Brain

In the study on the pathogenesis of viral encephalitis, the infection method is critical. The first of the two main infectious routes to the brain is the hematogenous route, which involves infection of the endothelial cells and pericytes of the brain. The second is the intracerebroventricular (ICV) route. Once within the central nervous system (CNS), viruses may spread to the subarachnoid space, meninges, and choroid plexus via the cerebrospinal fluid. In experimental models, the earliest stages of CNS viral distribution are not well characterized, and it is unclear whether only certain cells are initially infected. Here, we have analyzed the distribution of cytomegalovirus (CMV) particles during the acute phase of infection, termed primary viremia, following ICV or intravascular (IV) injection into the neonatal mouse brain. In the ICV injection model, 5 &#181;l of murine CMV (MCMV) or fluorescent microbeads were injected into the lateral ventricle at the midpoint between the ear and eye using a 10-&#181;l syringe with a 27 G needle. In the IV injection model, a 1-ml syringe with a 35 G needle was used. A transilluminator was used to visualize the superficial temporal (facial) vein of the neonatal mouse. We infused 50 &#181;l of MCMV or fluorescent microbeads into the superficial temporal vein. Brains were harvested at different time points post-injection. MCMV genomes were detected using the in situ hybridization method. Fluorescent microbeads or green fluorescent protein expressing recombinant MCMV particles were observed by fluorescent microscopy. These techniques can be applied to many other pathogens to investigate the pathogenesis of encephalitis.

Here, we describe a simple method of intracerebroventricular and intravascular injection of viral particles or fluorescent microbeads into the neonatal mouse brain. The localization pattern of the virus and nanoparticles could be detected by microscopic evaluation or by in situ hybridization.

In the study on the pathogenesis of viral encephalitis, the infection method is critical. The first of the two main infectious routes to the brain is the ...

Mouse Microsurgery Infusion Technique for Targeted Substance Delivery into the CNS via the Internal Carotid Artery

Animal models of central nervous system (CNS) diseases and, consequently, blood-brain barrier disruption diseases, require the delivery of exogenous substances into the brain. These exogenous substances may induce injurious impact or constitute therapeutic strategy. The most common delivery methods of exogenous substances into the brain are based on systemic deliveries, such as subcutaneous or intravenous routes. Although commonly used, these approaches have several limitations, including low delivery efficacy into the brain. In contrast, surgical methods that locally deliver substances into the CNS are more specific and prevent the uptake of the exogenous substances by other organs. Several surgical methods for CNS delivery are available; however, they tend to be very traumatic. Here, we describe a mouse infusion microsurgery technique, which effectively delivers substances into the brain via the internal carotid artery, with minimal trauma and no interference with normal CNS functionality.

Mouse Microsurgery Infusion Technique for Targeted Substance Delivery into the CNS via the Internal Carotid Artery

The present protocol describes a mouse microsurgery infusion technique, which effectively delivers substances directly into the brain via the internal carotid artery.

Animal models of central nervous system (CNS) diseases and, consequently, blood-brain barrier disruption diseases, require the delivery of exogenous ...

Cannula Implantation into the Cisterna Magna of Rodents

Cisterna magna cannulation (CMc) is a straightforward procedure that enables direct access to the cerebrospinal fluid (CSF) without operative damage to the skull or the brain parenchyma. In anesthetized rodents, the exposure of the dura mater by blunt dissection of the neck muscles allows the insertion of a cannula into the cisterna magna (CM). The cannula, composed either by a fine beveled needle or borosilicate capillary, is attached via a polyethylene (PE) tube to a syringe. Using a syringe pump, molecules can then be injected at controlled rates directly into the CM, which is continuous with the subarachnoid space. From the subarachnoid space, we can trace CSF fluxes by convective flow into the perivascular space around penetrating arterioles, where solute exchange with the interstitial fluid (ISF) occurs. CMc can be performed for acute injections immediately following the surgery, or for chronic implantation, with later injection in anesthetized or awake, freely moving rodents. Quantitation of tracer distribution in the brain parenchyma can be performed by epifluorescence, 2-photon microscopy, and magnetic resonance imaging (MRI), depending on the physico-chemical properties of the injected molecules. Thus, CMc in conjunction with various imaging techniques offers a powerful tool for assessment of the glymphatic system and CSF dynamics and function. Furthermore, CMc can be utilized as a conduit for fast, brain-wide delivery of signaling molecules and metabolic substrates that could not otherwise cross the blood brain barrier (BBB).

Here we describe a protocol to perform cisterna magna cannulation (CMc), a minimally invasive way to deliver tracers, substrates and signaling molecules into the cerebrospinal fluid (CSF). Combined with different imaging modalities, CMc enables glymphatic system and CSF dynamics assessment, as well as brain-wide delivery of various compounds.

Cisterna magna cannulation (CMc) is a straightforward procedure that enables direct access to the cerebrospinal fluid (CSF) without operative damage to ...

Transplantation of Chemogenetically Engineered Cortical Interneuron Progenitors into Early Postnatal Mouse Brains

Neuronal development is regulated by a complex combination of environmental and genetic factors. Assessing the relative contribution of each component is a complicated task, which is particularly difficult in regards to the development of &#947;-aminobutyric acid (GABA)ergic cortical interneurons (CIs). CIs are the main inhibitory neurons in the cerebral cortex, and they play key roles in neuronal networks, by regulating both the activity of individual pyramidal neurons, as well as the oscillatory behavior of neuronal ensembles. They are generated in transient embryonic structures (medial and caudal ganglionic eminences -&#160;MGE and CGE) that are very difficult to efficiently target using in utero electroporation approaches. Interneuron progenitors migrate long distances during normal embryonic development, before they integrate in the cortical circuit. This remarkable ability to disperse and integrate into a developing network can be hijacked by transplanting embryonic interneuron precursors into early post-natal host cortices. Here, we present a protocol that allows genetic modification of embryonic interneuron progenitors using focal ex vivo electroporation. These engineered interneuron precursors are then transplanted into early post-natal host cortices, where they will mature into easily identifiable CIs. This protocol allows the use of multiple genetically encoded tools, or the ability to regulate the expression of specific genes in interneuron progenitors, in order to investigate the impact of either genetic or environmental variables on the maturation and integration of CIs.

Here we present a protocol, designed to use chemogenetic tools to manipulate the activity of cortical interneuron progenitors transplanted into the cortex of early postnatal mice.

Neuronal development is regulated by a complex combination of environmental and genetic factors. Assessing the relative contribution of each component is ...

Delivery of Antibodies into the Murine Brain via Convection-enhanced Delivery

Convection-enhanced delivery (CED) is a neurosurgical technique enabling effective perfusion of large brain volumes using a catheter system. Such an approach provides a safe delivery method by-passing the blood brain barrier (BBB), thus allowing treatment with therapeutics with poor BBB-permeability or those for which systemic exposure is not desired, e.g., due to toxicity. CED requires optimization of the catheter design, injection protocol, and properties of the infusate. With this protocol we describe how to perform CED of a solution containing up to 20 &#181;g of an antibody into the caudate putamen of mice. It describes preparation of step catheters, testing them in vitro and performing the CED in mice using a ramping injection program. The protocol can be readily adjusted for other infusion volumes and can be used for injecting various tracers or pharmacologically active or inactive substances, including chemotherapeutics, cytokines, viral particles, and liposomes.

Convection-enhanced delivery (CED) is a method enabling effective delivery of therapeutics into the brain by direct perfusion of large tissue volumes. The procedure requires the use of catheters and an optimized injection procedure. This protocol describes a methodology for CED of an antibody into a mouse brain.

Convection-enhanced delivery (CED) is a neurosurgical technique enabling effective perfusion of large brain volumes using a catheter system. Such an ...

How to Obtain Reliable Visual Event-related Potentials in Newborns

The present study discusses the characteristics of visual event-related potentials (VEPs) and outlines methodological steps for obtaining reliable measurements in newborns. Obtaining high-quality, reliable VEPs is crucial for the early detection of abnormal development of the central nervous system in at-risk newborns, and for implementing successful early interventions. Recommendations are based on a previous study which showed that when post-conceptional age, polysomnography-identified sleep stages, and light-emitting diodes (LEDs) googles as the luminous source are controlled, no more than 4 repetitions of VEP averages are required to obtain replicable recordings, variability decreases, and reliable VEPs can be obtained. By controlling for these sources of variability and using statistical analyses, we were able to clearly and reliably identify the amplitude and latency of three main components (NII, PII and NIII) present in 100% of newborns (n = 20) during active sleep. Recording VEPs during awake states, quiet sleep and transitional sleep is not recommended because VEP morphology may differ significantly from one average to the next, leading to the risk of misleading clinical prognoses. Moreover, it is easier to obtain VEPs during active sleep because this state can be clearly and reliably identified at this stage of development, sleep cycles are short enough to allow measurements to be taken in a reasonable time, and the method does not require new o expensive equipment.

Several important points for obtaining high-quality reliable visual evoked potentials (VEPs) in newborns while minimizing variability and the risk of misleading prognoses are presented.

The present study discusses the characteristics of visual event-related potentials (VEPs) and outlines methodological steps for obtaining reliable ...

An Implantable System For Chronic In Vivo Electromyography

Electromyography (EMG) measures the muscle response to electrical stimulation or spontaneous activity of motor units and plays an important role in assessing neuromuscular function. Chronic recording of EMG activity reflecting a muscle&#8217;s reinnervation status after nerve injury has been limited, due to the invasive nature of traditional EMG recording techniques. In this regard, an implantable system is designed for long-term, in vivo EMG recording and nerve stimulation. It has been applied and tested in a study on reinnervation of laryngeal muscles. This system consists of 1) two bipolar electrode nerve cuffs and leads for stimulating each of two nerves: the recurrent laryngeal nerve (RLN) and internal branch of the superior laryngeal nerve (SLN); 2) two EMG recording electrodes and leads for each of the two laryngeal muscles: posterior cricoarytenoid (PCA) muscle and thyroarytenoid-lateral cricoarytenoid (TA-LCA) muscle complex; and 3) a skin receptacle interfacing all implanted lead terminals to an external recording preamplifier and stimulator using a connection cable. The wire leads are Teflon-coated, multi-filament, type 316 stainless steel. They are coiled and can stretch during body movement of the awake animal to prevent lead breakage and electrode migration. This system is implanted during an aseptic surgery. Afterwards, baseline EMG recordings are performed before the RLN is transected in the second surgery to study muscle reinnervation. Throughout the study, multiple physiological sessions are conducted in the anesthetized animal to obtain evoked and spontaneous EMG activity that reflects the reinnervation status of laryngeal muscles. The system is compact, free of infection over the course of the study, and highly durable. This implantable system can provide a reliable platform for research in which long-term recording or nerve stimulation is required in an anesthetized or freely moving animal.&#160;

Presented here is a protocol for the manufacturing of an implantable system for in vivo chronological recording of evoked and spontaneous electromyographic potentials. The system is applied to the investigation of reinnervation of laryngeal muscles following nerve injury. &#160;&#160;

Electromyography (EMG) measures the muscle response to electrical stimulation or spontaneous activity of motor units and plays an important role in ...