Protocols

• 7-AAD on Fixed Cells
  Stain dead cells and then fix them so that only the dead cells keep the 7-AAD
  
  
• Adherent Cell DNA Staining
  Only use if the other protocols fail. The samples are stored before staining, but it's more complicated.
  
  
• Antibody Staining
  General Antibody Staining Protocol
  
  
• Apoptosis with PI Only
  To stain cells with only PI to measure apoptosis (Quick & Dirty)
  
  
• Cell Cycle with PI
  First choice protocol to stain most cell types with Propidium Iodide (PI) to measure DNA content
  
  
• DNAse I
  DNAse treatment of cells that tend to clump
  
  
• Ethidium Monoazide (EMA)
  Stain dead cells and then be able to fix them, as with the 7-AAD above.
  
  
• GFP and DNA
  Determine the cell cycle of GFP positive and/or negative cells. Unfortunately, GFP usually has a 48 half-life!
  
  
• Isolated Nuclei DNA Staining
  A quick way to stain suspension or adherent cells without storage
  
  
• Nuclear Antigen and DNA Content with PI (2)
  Use one of these protocols to stain cells for a nuclear antigen and DNA content
  
  
• Surface Antigen and DNA Content with PI
  To stain surface antigens and measure DNA Content
  
  
• Sorting Preparation
  This will help you prepare for sorting & estimate yields
  
  
• Titration
  To get the most out of your antibodies, TITRATE THEM
  
  
• Yeast Cell Cycle
  These dyes work better than Propidium Iodide:
  • SYTOX Green
  • SYBR Green I

Other Protocols:

• 20X Phosphate Buffered Saline (PBS) Stock Solution
• Erythrocyte Lysis
• Leukocytes via Dextran Sedimentation
• Para-Formaldehyde Recipe

Links to Other Protocol Pages:

• Current Protocols in Cytomety (UCSD Only)
• Cytometry Techniques and Protocols (Purdue University)
• Imperial Cancer Research Fund FACS Laboratory (UK)
• Molecular Probes Handbook (Invitrogen)
• UCLA Flow Cytometry Core
• University of Florida Flow Cytometry Core Sample Preparation Protocols
• Yeast Cell Cycle (Susan Forsburg at the Salk Institute)
  

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Dennis J. Young, UCSD Cancer Center Flow Cytometry Core Facility