Purification of retinal ganglion cells from differentiation through adult via immunopanning and low-pressure flow cytometry

Sean M Riordan, Afnan M Aladdad, Kiran J McLoughlin, Karl E Kador*

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapter (peer-reviewed)peer-review

Abstract

The isolation and culturing of rodent retinal ganglion cells (RGC) is a key step in studying the function and cellular response of this crucial cell type. Typical methods used for isolation of RGCs include immunopanning or magnetic bead separation with antibodies targeting RGC specific protein markers. However, in developmental research, many of the most common markers, such as Thy-1, are not expressed in early stages of development. To help study these crucial early stage RGCs, we have developed a novel method that utilizes a transgenic mouse with a GFP tag on the protein BRN3 and a low-pressure fluorescence-activated cell sorter (FACS) system.
Original languageEnglish
Title of host publicationRetinal ganglion cells. Methods and protocols
EditorsBen Mead
PublisherSpringer
Pages11-24
Number of pages14
ISBN (Electronic)9781071634097
ISBN (Print)9781071634110
DOIs
Publication statusPublished - 10 Aug 2023

Publication series

NameMethods in Molecular Biology
Volume2708
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Mice
  • Retinal Ganglion Cells - metabolism
  • Retinal Ganglion Cells (RGC)
  • Mice, Transgenic
  • Fluorescence Activated Cell Sorting (FACS)
  • Antibodies - metabolism
  • Cell Differentiation
  • Immunopanning
  • Flow Cytometry
  • Brn3
  • Animals

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