Comparative investigation for auto-fluorescence sample detection by evaluating conventional and spectral flow cytometers performance

Abstract

Objective To compare the performance of conventional and spectral flow cytometers in detecting auto-fluorescence samples. Methods Single cell samples with auto-fluorescence were prepared and stained by annexin V(AV)binding and propidium iodure (PI) to detect apoptosis. The same auto-fluorescence sample was run on conventional and spectral flow cytometers side by side. Results By avoid using FITC in conventional cytometers, the better result was gotten in detecting auto-fluorescence samples. Full spectrum cytometry presented better resolution even there was spectral overlap. It was easy to discriminate the signals derived from auto-fluorescence and FITC. The protocol of annexinV binding and PI staining to analyze apoptosis was optimized. Conclusions Spectral flow cytometry shows relatively excellent performance under certain conditions in detecting of apoptosis derived from auto-fluorescence samples.

Keywords

• auto-fluorescence|flow cytometry|spectrum|fluorescein isothiocyanate(fitc)