Sort Cells Using Data In Real Time

Researchers working in flow cytometry require the highest-quality data from every sample in order to achieve increasingly challenging scientific goals. The demand for higher-resolution, multidimensional data has amplified interest in spectral data unmixing and analysis as compared to the traditional method of compensation for spectral spillover. Investigators frequently find it difficult or impossible to resolve dim, rare, or highly autofluorescent populations using standard compensation. Although some companies offer spectral analysis and post-acquisition sort solutions, workflows are cumbersome, and experiments consume large volumes of precious sample. Despite the shortcomings of current options, researchers still desire the power of spectral data handling. The Invitrogen™ Bigfoot Spectral Cell Sorter was developed with innovative hardware and software specifically to sort cells using spectrally unmixed data in real time.

However, many instruments that are produced today are uniquely configured to primarily perform spectral analysis, with limited options to perform the accepted, reliable, and trusted technique of compensation, which some researchers still want due to familiarity or for comparison to other methods. The Bigfoot Spectral Cell Sorter’s acquisition and sorting application, Sasquatch Software (SQS), allows the operator to sort and analyze either spectrally unmixed data or traditional compensated data.

In this paper, we will demonstrate the capability of the Bigfoot Spectral Cell Sorter using a fifteen-color panel. This panel was selected to show proof of principle for this cutting-edge technique of combining spectral workflow and multiparameter six-way cell sorting.