CytoFlex Flow Cytometer Application Notes

4. Reagents to add to tubes:

No Ab, no ADP (tube 1)

CD62P, no ADP (tubes 2)

CD62P + ADP (tubes 3)

PBS

45 μ L

34 μ L

34 μ L

Antibody

-

5 μ L

5 μ L

ADP

-

-

6 μ L

5. Add 5 µ L of whole blood to each tube, swirl gently to mix then incubate 30 min at room temperature in the dark.

6. Add 750 µ L filtered 1x PBS to each tube.

7. Acquire data as soon as possible.

Results

No ADP: All Events

10 3 10 4 10 5 10 6 SSC-A

Platelets(8.21 %)

10 4

10 5

0 10 3

10 6

FSC-A

Overlay of platelets stained with CD62P-PE untreated (green) and treated with ADP (red).

Data Acquisition and Analysis

Initial Hardware Settings

Approximate gains:

• FSC (log scale) - 40

• SSC (log scale) - 90

• FL2 (log scale) – 290

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