|
Purpose:
|
The distribution of fluorescence signals measured with flow cytometry can be influenced by several factors, including qualitative and quantitative properties of the used fluorochromes, optical properties of the detection system, as well as the variability within the analyzed cell population itself. Most of the single cell samples prepared from in vitro cultures or clinical specimens contain a variable cell cycle component. Cell cycle, together with changes in the cell size, are two of the factors that alter the functional properties of analyzed cells and thus affect the interpretation of obtained results. |
|
Conclusion:
|
Here, we describe the association between cell cycle status and cell size, and the variability in the distribution of fluorescence intensity as determined with flow cytometry, at population scale. We show that variability in the distribution of background and specific fluorescence signals is related to the cell cycle state of the selected population, with the 10% low fluorescence signal fraction enriched mainly in cells in their G0/G1 cell cycle phase, and the 10% high fraction containing cells mostly in the G2/M phase. We, therefore, advise using caution and additional experimental validation when comparing populations defined by fractions of the same size at both ends of fluorescence signal distribution to avoid biases caused by the effect of cell cycle and cell size.
|
|
Funding:
|
This work was supported by the Czech Science Foundation, grant nr. 20-22984S and 21-11585S; the Ministry of Health of the Czech Republic, grant nr. 18-08-00245, NU20J-07-00028; the European Structural and Investment Funds, Operational Program Research, Development and Education – „Preclinical Progression of New Organic Compounds with Targeted Biological Activity” (Preclinprogress) - CZ.02.1.01/0.0/0.0/16_025/0007381; The project National Institute for Cancer Research (Programme EXCELES, ID Project No. LX22NPO5102) - Funded by the European Union – Next Generation EU. JR is supported by the Terri Brodeur Breast Cancer Foundation and MSKCC Support Grant P30 CA008748.
|
