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Method Article
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July 8th, 2025
Source: Bączyk, M., et al. In Vivo Intracellular Recording of Type-Identified Rat Spinal Motoneurons During Trans-Spinal Direct Current Stimulation. J. Vis. Exp. (2020).
This video describes the in vivo intracellular recording of rat motoneurons under trans-spinal direct current stimulation. The protocol describes how to measure membrane properties and record the rhythmic firing of motoneurons before, during, and after anodal or cathodal polarization of the spinal cord.
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
NOTE: Each participant involved in the procedure has to be properly trained in basic surgical procedures and has to have a valid license for performing animal experiments.
1. Anesthesia and premedication
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| Name | Company | Catalog Number | Comments |
|---|---|---|---|
| Durgs and solutions | - | - | - |
| Atropinum sulfuricum | Polfa Warszawa | - | - |
| Glucose | Merck | 346351 | - |
| NaHCO3 | Merck | 106329 | - |
| Pancuronium Jelfa | PharmaSwiss/Valeant | - | Neuromuscular blocker |
| Pentobarbital sodium | Biowet Pu awy Sp. z o.o | - | Main anesthetic agent |
| Pottasium citrate | Chempur | 6100-05-06 | - |
| Tetraspan | Braun | - | HES solution |
| Surgical equipment | - | - | - |
| 21 Blade | FST | 10021-00 | Scalpel blade |
| Cauterizer | FST | 18010-00 | - |
| Chest Tubes | Mila | CT1215 | - |
| Dumont #4 Forceps | FST | 11241-30 | Muscle forceps |
| Dumont #5 Forceps | FST | 11254-20 | Dura forceps |
| Dumont #5F Forceps | FST | 11255-20 | Nerve forceps |
| Dumont #5SF Forceps | FST | 11252-00 | Pia forceps |
| Forceps | FST | 11008-13 | Blunt forceps |
| Forceps | FST | 11053-10 | Skin forceps |
| Hemostat | FST | 13013-14 | - |
| Rongeur | FST | 16021-14 | For laminectomy |
| Scissors | FST | 15000-08 | Vein scissors |
| Scissors | FST | 15002-08 | Dura scissors |
| Scissors | FST | 14184-09 | For trachea cut |
| Scissors | FST | 104075-11 | Muscle scissors |
| Scissors | FST | 14002-13 | Skin scissors |
| Tracheal tube | - | - | Custom made |
| Vein catheter | Vygon | 1261.201 | - |
| Vessel cannulation forceps | FST | 18403-11 | - |
| Vessel clamp | FST | 18320-11 | For vein clamping |
| Vessel Dilating Probe | FST | 10160-13 | For vein dissection |
| Sugrgical materials | - | - | - |
| Gel foam | Pfizer | GTIN 00300090315085 | Hemostatic agent |
| Silk suture 4.0 | FST | 18020-40 | - |
| Silk suture 6.0 | FST | 18020-60 | - |
| Equipment | - | - | - |
| Axoclamp 2B | Molecular devices | discontinued | Intracellular amplifier/ new model Axoclamp 900A |
| CapStar-100 End-tidal CO2 Monitor | CWE | 11-10000 | Gas analyzer |
| Grass S-88 | A-M Systems | discontinued | Constant current stimulator |
| Homeothermic Blanket Systems with Flexible Probe | Harvard Apparatus | 507222F | Heating system |
| ISO-DAM8A | WPI | 74020 | Extracellular amplifier |
| Microdrive | - | - | Custom made/replacement IVM/Scientifica |
| P-1000 Microelectrode puller | Sutter Instruments | P-1000 | Microelectrode puller |
| SAR-830/AP Small Animal Ventilator | CWE | 12-02100 | Respirator |
| Support frame | - | - | Custom made/replacement lab standard base 51601/Stoelting |
| Spinal clamps | - | - | Custom made/replacement Rat spinal adaptor 51695/Stoelting |
| TP-1 DC stimulator | WiNUE | - | tsDCS stimulator |
| Miscellaneous | - | - | - |
| 1B150-4 glass capillaries | WPI | 1B150-4 | For microelectrodes production |
| Cotton wool | - | - | - |
| flexible tubing | - | - | For respirator and CO2 analyzer connection |
| MicroFil | WPI | MF28G67-5 | For filling micropipettes |
| Silver wire | - | - | For nerve electrodes |
