Identification of Mouse Retinal Ganglion Cell Subtypes Through Fluorescent Tracing

Abstract

Source: Laboissonniere, et. al. Single-cell RNA-Seq of Defined Subsets of Retinal Ganglion Cells. J. Vis. Exp. (2017).

This video demonstrates a fluorescent tracing technique to identify retinal ganglion cell (RGC) subtypes in a transgenic mouse retina. The process involves securing the isolated retina in a recording chamber, maintaining cell viability with oxygenated perfusion, and targeting fluorescently labeled RGCs using a pipette with a tracer. By analyzing dendritic morphology and stratification, the method enables classification of distinct RGC subtypes.

Protocol

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

Results

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Diethyl pyrocarbonate (DEPC)	Sigma Aldrich	D5758
Alexa Fluor 594 Hydrazide	Invitrogen	A10442
Petri dish (35mm diameter)	Thermo Fisher Scientific	153066
Ophthalmologic scissors	Fine Science Tools	15000-00
#5 Forceps	Fine Science Tools	11252-30
Glass Micropipette	Sutter	BF120-69-10
Micropipette Puller	Sutter		P-1000 horizontal pipette puller
1mL syringe	Fisher Scientific	14-823-2F