16 color panel on Human Peripheral Blood
Application data using 2 lasers
Fluorochromes: 488nm Laser
Fluorochromes: 405nm Laser
| HLA-DR | Pacific Blue |
| CD38 | BV421 |
| CD45RA | BV510 |
| CD33 | BV570 |
| CD16 | BV605 |
| CD11b | BV650 |
| CD11c | BV711 |
| CD14 | BV785 |
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).
16 color stained sample of Human Peripheral Blood
Application data using 3 lasers
Fluorochromes: 405nm Laser
Fluorochromes: 488nm Laser
| CD3 | Alexa Fluor 488 |
| CD8 | PE |
| CD4 | PE Dazzle 594 |
| CD24 | PE Alexa Fluor 610 |
| CD19 | PE Cy5 |
| CD20 | PE Cy5.5 |
| CD13 | PerCP efluor® 710 |
| CD45 | PE Cy7 |
Fluorochromes: 638nm Laser
| CD38 | APC |
| C11c | Alexa Fluor 700 |
| CD45RA | APC-Cy7 |
Example of the detection of several important cell populations using sixteen fluorochromes excited by three lasers. Unlike conventional flow cytometers that can detect up to five off the blue, in this example we demonstrate that with the SP6800 eight fluorochromes can be excited off the blue with clear sample resolution. Even highly overlapping fluorchromes such as PE-Cy5 and PE-Cy5.5 can be resolved.
Application Data: Brilliant Violet Dyes
Spectral Analysis allows multi-laser excited fluorochromes to be run without using a complicated compensation matrix.
| BV421 | BV605 |
| BV510 | BV650 |
| BV570 | BV711 |
| BV785 |
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
Example 5
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)
