Establishing a Whole-Cell Voltage-Clamp Configuration for Electrophysiological Recordings in Brain Slices

Abstract

Source: Sunstrum, J. K., et al. Evaluation of synaptic multiplicity using whole-cell patch-clamp electrophysiology. J. Vis. Exp. (2019)

This video demonstrates positioning a fine-pointed pipette near brain slices, forming a seal with a neuronal cell. Suction breaks the cell membrane, thereby creating a whole-cell patch clamp that enables current measurements for electrophysiology experiments.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.            

1. Solutions

1. Slicing solution​
   1. Refer to Table 1 for the composition of the slicing solution.
   2. Prepare a 20x stock solution in advance and store it at 4 °C for up to 1 month.
   3. For the 1x slicing solution, dissolve NaHCO₃, glucose, and sucrose in ddH₂O and add the 20x stock. Ensure the osmolarity is betw....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
1 ml syringe	BD	309659
10 blade	Fisher Scientific/others	35698
22 blade	VWR/others	21909-626
22 uM syringe filters	Milipore	09-719-000
Adson foreceps	Harvard Instruments	72-8547
Angled sharp scissors	Harvard Instruments	72-8437
Clampex	Molecular Devices	pClamp 10
Double edge blade	VWR	74-0002
Filter paper	Sigma/others	1001090
Fine paintbrush	Fisher/various	15-183-35/various
Gas Dispersion Tube	VWR	LG-8680-120
Isoflurane	Fresenius Kabi/others	M60303
Krazy glue	various	various
Mini analysis	Synaptosoft	MiniAnalysis 6
Osmomoter	Wescor Inc	Model 5600
Parafilm	Sigma	PM-996
Pasteur pipette	VWR	14672-200
ph meter	Mettler Toledo	FE20-ATC
Rubber bulb	VWR	82024-550
Scalpel handle No. 3	Harvard Instruments	72-8350
Scalpel handle No. 4	Harvard Instruments	72-8356
Single edge blade	VWR	55411-050
Vibratome slicer	Leica	VT1200S
Water Purification System	Millipore	Milli-Q Academic A10
Well plate lid	Fisher/various	07-201-590/various
CaCl2 2H2O	Sigma	C7902
CdCl2	sigma	202908
EGTA	Sigma	E3889
glucose	Sigma	G5767
HEPES	Sigma	H3375
K2-ATP	Sigma	A8937
KCl	Sigma	P9333
K-gluconate	Sigma	G4500
MgCl2 6H2O	Sigma	M2670
Molecular biology grade water	Sigma	W4502-1L
Na3GTP	Sigma	G8877
NaCl	Bioshop	SOD001.1
Na-gluconate	Sigma	S2054
NaH2PO4	Sigma	71504
NaHCO3	Sigma	S6014
Picrotoxin	sigma	P1675
SrCl	Sigma	255521
sucrose	Bioshop	SUC507.1