In flow cytometry fluorescent light emitted from each fluorochromes is distributed over some part of visible spectrum. Cells are often stained with more than one fluorochrome, each having its own spectral distribution. Usually each fluorescent emission spectra overlap. In conventional filtering systems, overlapping fluorescence is subtracted using color compensation, so less light is collected.
Instead, spectral flow cytometers sum the fluorescence together and then use unmixing technology to mathematically separate the colors. In the second video this feature is better shown by an intuitive figure with colored flasks!
The spectral flow streamlines workflow, including panel design, improves visualization of autofluorescence and yields better data by collecting the entire visual spectrum of light. As shown in the figure below, we can see the difference between Spectral Flow Cytometry and conventional flow cytometry.
In doing so it enhances dim signal detection for better visualization of rare populations, fluorescent proteins, and fluorochromes excited by multiple lasers.