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  • DavidLeclerc

ChUG #8 - Dealing with autofluorescence in spectral flow cytometry

We host Monica Delay, US Manager for Technical Application Support at Cytek Biosciences, for a discussion on autofluorescence (AF) and the tools to deal with it in Spectral Flow Cytometry. We mean to provide an introduction to this concept, and links to further your understanding. Scroll below to find a list of webinars and publications dealing with AF.


Upcoming Webinar by Diana Bonilla Escobar: August 4, 2021

If you are an owner or user of a Cytek Aurora, you can find additional information in the user forum within the archives of both the April 2021 and January 2021 User Group Meetings:

Recorded Webinar by Alexis Gonzalez “Just Because You Don’t See It Doesn’t Mean It Isn’t There: Improved Assay Resolution with Autofluorescence Extraction”:

(UChicago) Video tutorial for setting up unstained controls in SpectroFlo:

(Cytek) Video tutorial for the SpectroFlo autofluorescence extraction feature:

(Cytek) Video tutorial for the SpectroFlo group specific unstained control feature:


- Roederer M, Murphy RF. Cell-by-cell autofluorescence correction for low signal-to-noise systems: application to epidermal growth factor endocytosis by 3T3 fibroblasts. Cytometry. 1986 Nov;7(6):558-65. doi: 10.1002/cyto.990070610. PMID: 3096673.

- Alberti S, Parks DR, Herzenberg LA. A single laser method for subtraction of cell autofluorescence in flow cytometry. Cytometry. 1987 Mar;8(2):114-9. doi: 10.1002/cyto.990080203. PMID: 3556100.

- Olson RJ, Zettler ER, Anderson OK. Discrimination of eukaryotic phytoplankton cell types from light scatter and autofluorescence properties measured by flow cytometry. Cytometry. 1989 Sep;10(5):636-43. doi: 10.1002/cyto.990100520. PMID: 2776580.

- Corsetti JP, Sotirchos SV, Cox C, Cowles JW, Leary JF, Blumburg N. Correction of cellular autofluorescence in flow cytometry by mathematical modeling of cellular fluorescence. Cytometry. 1988 Nov;9(6):539-47. doi: 10.1002/cyto.990090606. PMID: 3208620.

- Mosiman VL, Patterson BK, Canterero L, Goolsby CL. Reducing cellular autofluorescence in flow cytometry: an in situ method. Cytometry. 1997 Jun 15;30(3):151-6. PMID: 9222101.

- Shilova ON, Shilov ES, Deyev SM. The effect of trypan blue treatment on autofluorescence of fixed cells. Cytometry A. 2017 Sep;91(9):917-925. doi: 10.1002/cyto.a.23199. Epub 2017 Aug 30. PMID: 28857464.

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