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Cell Cycle Analysis and GFP Expression of Zebrafish Embryos by FACS
Published: Feb 20, 2012 DOI: 10.21769/BioProtoc.185 Views: 17175
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Abstract
Cell cycle analysis in combination of GFP expression has been well used to study the cell cycle distribution of GFP labeled cells. This protocol is a method to analyze the cell cycles of GFP marked cells from zebrafish embryos.
Materials and Reagents
- Phosphate buffered saline (PBS) (Life Technologies, Invitrogen™, catalog number: 14040 )
- Fetal bovine serum (FBS)
- Ethanol
- PFA (USB)
- EtOH
- Formaldehyde
- RNaseA
- Hoechst 33342 solution (Life Technologies, Molecular Probes, catalog number: H3570 )
- 40 μm cell strainer (BD Biosciences, catalog number: 352340 )
- Polystyrene FACS tube (BD Biosciences, Falcon®, catalog number: 352054 )
- 50 ml Falcon tube (BD Biosciences, catalog number: 352070 )
- PI solution (see Recipes)
- PBS + 5% FBS (see Recipes)
Equipment
- FACS
- Standard tabletop centrifuges
- Water bath
- Polystyrene FACS tube
- Cell strainer
Procedure
Category
Cell Biology > Cell imaging > Fluorescence
Cell Biology > Cell-based analysis > Flow cytometry
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