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Optimizing Reference Controls for Multicolor Flow Cytometry

Best practices for preventing compensation or unmixing errors.

Created in collaboration with
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The quality of reference controls is critical for generating accurate compensation or unmixing matrices and correctly interpreting conventional or spectral flow cytometry data, respectively. Follow this checklist to help prevent common errors due to mismatch between reference and sample, differences in autofluorescence, inadequate gating, insufficient cell number, and use of untested and unvalidated cell surrogates.

Reference control checklist
Modified from © ISTOCK.COM, BGBLUE, RAMBO182, NADIINKO, PHOTOTECHNO; Designed by Ashleigh Campsall

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  1. Spectral flow cytometry. BD Biosciences. Accessed May 8, 2024.