Introduction to Spectral Flow Cytometry

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Below is an excerpt from an article published on the Sony Biotechnology blog on an introduction to spectral cytometry.
Read the full article here.

Introduction to Spectral Flow Cytometry

Spectral flow cytometry has been a part of the Sony Biotechnology portfolio since 2013, and the ID7000™ Spectral Cell Analyzer is the latest addition to our range. This innovative system enables user-friendly multiparametric detection with a number of key benefits, including improved flexibility in panel design, the ability to re-use spectral references, and tools for removal of autofluorescence. Below are some key areas where this spectral technology delivers advantages over conventional flow cytometry.

Spectral flow cytometry

Signals from all detected channels are used, regardless of the number of fluorochromes analyzed. The fluorescence intensity is additive, and sums up wherever the spectra overlap.

There is no signal subtraction. Instead, fluorescence from each cell is measured at many points throughout the spectrum to define the overall fluorescence

This spectrum includes the sum of the fluorescence from all of the fluorochromes, as well as autofluorescence. The spectral data is the fluorescence intensity measured by each of the detectors for each and every cell.
In spectral cell analyzers, each fluorochrome uses all the detection channels to produce a spectral emission signal. This signal is converted into a reference spectrum for each dye, which can then be used in spectral unmixing calculations. Using a WLS-based algorithm, spectral unmixing takes into account the actual noise detected in a sample to ensure accurate estimation of each dye’s intensity, and allows auto-fluorescence to be handled as a separate color.

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