Flow cytometry has long been considered a tool of hematologists and immunologists who primarily work with the hematopoetic system.  Flow cytometry is capable of performing the simultaneous detection of more than 20 parameters; however it requires the sample to be prepared into a single cell suspension. The need for a single cell suspension makes the sample preparation more intensive for cells derived from tissues. Many tissues are also highly autofluorescent, which results in background noise that limits detection of low levels of expression on conventional flow cytometers.

Researchers from the Institut Pasteur, Immunology Department in Paris France have published a study in PLoS One which simultaneously measured 21 parameters (19 fluorochromes) on cells derived from mouse embryonic heart and intestine. Both tissue types contain autofluorescent cells. With conventional cytometers, specific band pass filters must be carefully selected to enable compensation for spectral overlap. The more overlap between fluorescent dyes, the more difficult to perform conventional compensation. Furthermore, cells such as those described in the article have larger amounts of inherent autofluorescence complicating the data analysis. With the SP6800 a spectral unmixing algorithm is used to determine the abundance of signal from each fluorochrome. This allows for the separation of highly overlapping signals including GFP and FITC as well as effective management of autofluorescence. Directly comparing results on two conventional flow cytometers and the SP6800, only the SP6800 was capable of effective phenotypic resolution of the target populations.

References

Schmutz S, Valente M, Cumano A, Novault S "Spectral Cytometry Has Unique Properties Allowing Multicolor Analysis of Cell Suspensions Isolated from Solid Tissues." PLoS One 11(8): e0159961 2016. PubMed