Flow Cytometry/Cell Sorting
Transcript of Flow Cytometry/Cell Sorting
Flow Cytometry/Cell Sorting Shared Resource (FCSR)Karen Creswell, PhD, [email protected]
FCSR – Apoptosis Assays
• Sub G1 peak
• Annexin V (membrane flip-flop)
• TUNEL assay (DNA strand breaks)
• JC-1 staining (mitochondrial membrane potential)
• Caspase and caspase activity assays
FCSR: Stem Cell Assays
• Immunophenotyping
• Aldefluor analysis
• Side population (SP) detection
Any of these methods or a combination
of methods can be utilized to analyze
and sort stem cells
Aldefluor-A
SSC-A
100
101
102
103
104
0
256
512
768
1024
+DEAB
ALDH + 0.03 %
SSC-A
Aldefluor-A
SSC-A
100
101
102
103
104
0
256
512
768
1024
+DEAB
ALDH + 0.03 %
SSC-A
FCSR: Other Applications
•Cell Cycle
•Immunophenotyping
•Cytoxicity Assays
•Measurement of ROS
•Phagocytosis Assays
•Intracellular cytokine analysis
•Cell signaling antigen detection
•Fluorescent protein analysis (GFP, CFP, YFP, mCherry, etc.)
GG22MM GG00
GG11
ss
0 200 400 600 800 1000
GG00GG11
ss GG22 MM
DNA AnalysisDNA Analysis
DNA content
Count
2N2N 4N4N
Normal Cell Cycle Normal Cell Cycle
FCSR: Cell Sorting•Separation of cell populations
•Can sort up to 4 populations simultaneously•Can sort into 6-, 24, or 96-well plates
Shared Resources are partially supported by the Georgetown Howard University Center for Clinical and Translational Studies (GHUCCTS) 1UL1RR031975-01
In your publications, remember to mention support provided from the GHUCCTS.