Flow Cytometry/Cell Sorting Shared Resource (FCSR)Karen Creswell, PhD, Directorcreswelk@georgetown.edu202-687-2233

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.