Light-Evoked Postsynaptic Responses in Neurons of Mouse Retinal Slices

Abstract

Source: Hellmer, C. B., et al. Recording Light-evoked Postsynaptic Responses in Neurons in Dark-adapted, Mouse Retinal Slice Preparations Using Patch Clamp Techniques. J. Vis. Exp. (2015).

This video demonstrates the recording of light-evoked post-synaptic responses in a dark-adapted mouse retinal tissue slice preparation. The retinal slice is placed in a recording chamber, and a recording pipette is used to establish electrical continuity with a retinal ganglion cell. Light pulses are applied to generate excitatory post-synaptic potentials in the ganglion cells, which are recorded by the pipette.

Protocol

All procedures involving sample collection have been performed in accordance with the institute's IRB guidelines.

1. Preparation of Experimental Solution

1. Prepare the dissecting solution 1 day to 1 week before the actual experiment. Use a 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) buffer solution for retinal dissection because of its strong buffering ability at lower temperatures16. Mix all chemicals as follows (in mM): 115 NaCl, 2.5 KCl, 2.5 CaCl₂, 1.0 MgCl₂, 10 HEPES, 28 glucose. Adjust pH to 7.4 with NaOH. Keep the solution in a refrigerator up to 1 week.
2. The rec....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
mice (28-60 days old, male)	Jackson laboratory	C57BL/6J strain
Ames' medium powder	Sigma	A1420	excellent
infrared viewer	Night Owl Optics	NOBG1	It shows bright view. Focusing small objects is an issue.
infrared pocket scopes	B.E. Meyers	OWL Gen 3 NV pocketscope	excellent view
puller	Sutter	P-1000	excellent. Make consistent size pipettes.
dark box	Pelican	dark box	excellent
patch clamp system	Scientifica	slice scope 2000	Excellent setup. Most key components are included in one package. Micromanipulators are excellent.
amplifier	Molecular Devices	multiclamp 700B	Excellent and easy control.
acquiring software	Molecular Devices	pClamp software	Excellent and easy control.
light source (LED)	Cool LED	pE-2 4 channel system	Excellent
CCD camera	Q-imaging	Retiga 2000	Excellent
Faraday cage	handmade