Núria Crua· Postdoc· University of Lausanne (UNIL)
Published Jul 31, 2026~30m total· 0 contributions
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JNAR-ID: JNAR-74736672
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PublishedLast updated Jul 31, 2026
Objective
Prepare single-color compensation controls for flow cytometry using antibody capture beads.
Classification
Sample typeSynthetic Sample
TechniqueFlow Cytometry / FACS
Experimental contextIn vitro
Reproducibility status
Reproducible
No reproducibility blockers or risks reported. The protocol runs end-to-end.
10 healthy
⏱ Total 30 min1 section10 steps
Preparation and Staining
30 min
1
Vortex the compensation beads vigorously for 5–10 seconds to ensure a homogeneous suspension.
2
Add one drop of beads to each well of a 96-well round-bottom plate (one well per fluorochrome).
3
Centrifuge at 1,000 × g for 5 minutes at 4°C.5 min
4
Carefully discard the supernatant.
5
Prepare the antibody dilutions in separate tubes using FACS buffer. Add 100 µL of the appropriate antibody solution to each well (e.g., 1 µL antibody in 100 µL FACS buffer for a 1:100 dilution).
6
Incubate for 20 minutes at 4°C, protected from light. 20 min
7
Centrifuge at 1,000 × g for 5 minutes at 4°C.5 min
8
Discard the supernatant and resuspend the beads in 200 µL FACS buffer.
9
Wash the beads once with FACS buffer, centrifuge under the same conditions, discard the supernatant, and resuspend in an appropriate volume of FACS buffer for acquisition.
10
Acquire each single-color compensation control individually and save the resulting .fcs files for compensation matrix calculation.
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