SP6800 Spectral Analyzer

Application data using 2 lasers

Examples of the resolution of several important cell populations using only two lasers with the SP6800. Clear population resolution is obtained with highly overlapping fluorochomes such as PE-Cy5/PE-Cy5.5 (G) and Pacific Blue/BV421 (H).

Application Data: Brilliant Violet Dyes
Spectral Analysis allows multi-laser excited fluorochromes to be run without using a complicated compensation matrix.

Sample data from Human PBMCs stained with seven markers conjugated to Brilliant Violet dyes offered by Sony Biotechnology. D. All populations were clearly resolved including fluorochomes with significant spectral overlap such as BV605 and BV650 (D).

12-color Staining of Human Peripheral Blood Leukocytes
Example of Human Peripheral Blood Leukocytes was analyzed on the SP6800 and a BD LSRFortessa conventional flow cytometer. Spectral analysis was able to remove the autofluorescenece and use unmixing to separate each fluorochrome which resulted in clarity and resolution of each population in these density plots

Example 1

Example 2

Example 3

Example 4

Application Data: Dyes with issues
Common problems in flow cytometry occur when running fluorochromes with emission peaks that are too close to one another, multi-laser excitations, fluorescent proteins, and unstable tandems. The SP6800 is capable of analyzing all of these by looking at all photons from 420nm to 800nm and unmixing each unique spectral fingerprint.

Example 1: Pacific Blue (457nm emission) and Brilliant Violet 421 (422nm emission)

Example 2: PE Cy5 (excited with 488nm and 638nm lasers) and APC (excited with 638nm laser)

Example 3: Fluorescent GFP and YFP need no special bandpass filter set with the SP6800

Example 4: PE Cy7 tandem is prone to falling apart causing issues with resolution when using conventional flow cytometers. Since Spectral Analysis unmixes the spectral fingerprint, the SP6800 produces excellent resolution.