Paired Patch Clamp Recording of the Neuronal Connections in Acute Brain Slices

Abstract

Source: Qi, G., et al. Electrophysiological and Morphological Characterization of Neuronal Microcircuits in Acute Brain Slices Using Paired Patch-Clamp Recordings. J. Vis. Exp. (2015)

This video demonstrates the procedure to map synaptic connections by securing a brain slice, positioning pipettes to achieve whole-cell configurations, and stimulating presynaptic neurons to record postsynaptic responses, thereby elucidating neuronal connectivity.

Protocol

All procedures involving animal samples have been reviewed and approved by the appropriate animal ethical review committee.

1. Paired Patch-Clamp Recording and Biocytin Filling

Depending on the type of synaptic connection, three different approaches are used to find synaptically coupled neurons. If the connection probability is low (as can be expected for most excitatory connections), proceed as follows:

1. Neuronal connections with a low connectivity
   1. Fill the patch pipettes with an internal solution of the composition listed in Table....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Amplifier	HEKA	EPC 10 USB Triple	with 2-3 preamplifiers
Microscope	Olympus	BX51WI	with 2 camera ports and a 4× objective, a 40× water-immersion objective
Camera	TILL Photonics	VX55	infrared CCD camera
Workstation	Luigs & Neumann	Infrapatch 240	with a motorized x-y stage and a motorized focus axis for the microscope
Micromanipulator	Luigs & Neumann	SM-5	x-y-z manipulators for 2-3 preamplifiers
Faraday cage	Luigs & Neumann
Anti-vibration table	Newport Spectra-Physics
Patchmaster	HEKA
Microtome	Microm International	HM650V
Micropipette puller	HEKA	Sutter P-97
Neurolucida system	Microbrightfield		with Neurolucida and Neuroexplorer softwares