PUBLICATION
Preparing a Single Cell Suspension from Zebrafish Retinal Tissue for Flow Cytometric Cell Sorting of Müller Glia
- Authors
- Allan, K., DiCicco, R., Ramos, M., Asosingh, K., Yuan, A.
- ID
- ZDB-PUB-191127-9
- Date
- 2019
- Source
- Cytometry. Part A : the journal of the International Society for Analytical Cytology 97(6): 638-646 (Journal)
- Registered Authors
- Keywords
- Müller glia, dissociation, flow cytometric cell sorting, papain, zebrafish retina
- MeSH Terms
-
- Animals
- Cell Proliferation
- Flow Cytometry
- Neuroglia
- Retina*
- Zebrafish*
- PubMed
- 31769194 Full text @ Cytometry A
Citation
Allan, K., DiCicco, R., Ramos, M., Asosingh, K., Yuan, A. (2019) Preparing a Single Cell Suspension from Zebrafish Retinal Tissue for Flow Cytometric Cell Sorting of Müller Glia. Cytometry. Part A : the journal of the International Society for Analytical Cytology. 97(6):638-646.
Abstract
Preparation of a single cell suspension from solid tissue is vital for a successful flow cytometry experiment. We report a detailed and reproducible method to produce a quality cell suspension from the zebrafish retina. Zebrafish retinas, especially their Müller glia cells, are of particular interest for their inherent regenerative capacity, making them a useful model for regenerative medicine and cell therapy research. Here, we detail a papain-based dissociation that is gentle enough to keep cells intact, but strong enough to disrupt cell-cell and cell-matrix interactions to yield a cell suspension that produces clean and reliable flow cytometric cell sorting results. This procedure consistently results in over 90% viability and three populations of cells based on GFP expression. The dissociation procedure described herein has been optimized for the collection of Müller glia from Tg(apoe:gfp) zebrafish retinas; however, the overall process may be applicable to other cell types in the fish retina, additional flow cytometric techniques, or preparing cell suspensions from similar tissues. © 2019 International Society for Advancement of Cytometry.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping