Spectral Flow Cytometers
Overview and technology
Spectral Flow Cytometry is a new generation of flow cytometers, originally created by Sony Biotechnology, that collects the entire emission spectrum of fluorophores instead of just a band of the emission spectrum for each detector – as in done in conventional flow cytometry. Traditionally each fluorophore emission was detected by a separate detector thus “grabbing” this part of the spectrum using a bandpass optical filter.
With this methodology, if multiple fluorescence markers are to be detected simultaneously, careful planning of the fluorescent panel as well as complex compensation matrices are required.
Sony Biotechnology has dramatically innovated the detection method of FACS analyzers by collecting the entire spectrum of emitted light for each fluorophore, thus creating a spectral fingerprint for each marker that is unique. This method significantly simplifies the data collection and analysis process – thus allowing a much wider range of panels to be used, especially for high count multi-color experiments. The separation of the different colors is performed by a spectral unmixing algorithm, instead of compensation, which increases the sensitivity and quality of the data.
Use cases
Spectral flow cytometry is heavily used in the field of immunology, where there is a need for 10-40 markers simultaneously to analyze the different cell populations.
Spectral cytometry also enables the detection of the entire autofluorescent spectrum. For cells with high autofluorescence, this is used to either subtract the autofluorescence from the marker emission or as a fluorescent parameter of its own – to distinguish specific cell types and/or to indentify cell apoptosis and necrosis processes without additional markers.
Unique benefits:
Spectral cytometry increases simplicity, combination, and quantity of fluorescent panels, sensitivity, and quality of data over conventional flow cytometry.