Flow cytometry for quantification of retrogradely labeled retinal ganglion cells by Fluoro-Gold

Junjie Yang, Gülgün Tezel, Rajkumar V. Patil, Martin B. Wax

Research output: Contribution to journalArticlepeer-review

9 Scopus citations

Abstract

Purpose. To count retrogradely labeled retinal ganglion cells by Fluoro-Gold. Methods. Retinal ganglion cells were retrogradely labeled using bilateral injections of Fluoro-Gold into the superior colliculus. One week after injections, retinas were dissociated and immunolabeled using specific antibody against Fluoro-Gold. The Fluoro-Gold labeled cells were then counted using flow cytometry. Results. Flow cytometry revealed that approximately 7% of the dissociated retinal cells were ganglion cells retrogradely labeled by Fluoro-Gold (Fig. 1B). Based on the total count of retinal cells per eye, the total number of retinal ganglion cells was estimated at approximately 131,250 ± 2,542 per rat eye. The coefficient of variation of counts was calculated as 1.98%. Conclusions. The use of flow cytometry facilitates simple, reproducible and rapid quantification of virtually all of the retinal ganglion cells labeled by Fluoro-Gold in a single rat eye.

Original languageEnglish (US)
Pages (from-to)981-985
Number of pages5
JournalCurrent Eye Research
Volume21
Issue number6
DOIs
StatePublished - Dec 1 2000

Keywords

  • Anti-Fluoro-Gold antibody
  • Flow cytometry
  • Fluoro-Gold
  • Retinal ganglion cells
  • Retrograde labeling

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

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