Is spectral flow cytometry the solution to all problems?

Spectral flow cytometry has made a significant advance by expanding the number of parameters that can be evaluated simultaneously in a single panel. This has made it possible to explore the heterogeneity of cell populations present in our samples much more deeply. Furthermore, this technology has increased the resolution of labeling, as it offers more powerful detectors and the possibility of extracting autofluorescence.

However, even though it is an important milestone in the field of flow cytometry, it is not, by itself, the solution to all problems. It is still necessary to plan and optimize the panels (often with greater rigor than in conventional cytometry) to guarantee quality results.

In conventional flow cytometry, only a portion of the fluorescence emitted by a fluorochrome is detected in a specific channel (a channel is understood as the combination of optical filters that selects a narrow band of the emission spectrum and directs it to a detector). Therefore, since each channel selectively detects only a band of fluorescence, the number of fluorochromes that can be used simultaneously is limited by the number of channels available in the flow cytometer.

In spectral flow cytometry, a large portion of the fluorescence spectrum emitted by fluorochromes is detected. To achieve this, multiple detectors cover a wide range of wavelengths, allowing the determination of the spectral signature of each fluorochrome. In this way, it is possible to distinguish fluorochromes even when they have very similar emission spectra – which would be detected in the same channel in conventional flow cytometry and could not be used simultaneously.

We can see that this technology offers numerous benefits, but only when implemented following best practices. With that in mind, in the coming weeks we will be launching a series of posts called "10 Days, 10 Tips for Using Spectral Flow Cytometry." Follow along and learn more!

Series of posts: 10 days, 10 tips on spectral flow cytometry