Instrumentation - Flow cytometers and Cell sorters
|Flow cytometer Instrument||Laser||Marker||Comments|
|BD LSR Fortessa||4||17|
|Sony SP6800ZE Spectral Analyzer||3||20||full spectrum analyzer uses spectral unmixing instead of conventional compensation|
|Cell sorter Instrument||Laser||Marker||Comments|
|BD FACS Aria Fusion||4||18||Sort 4 cell populations simultaneously|
|BD FACS Aria III||5||21||Sort 4 cell populations simultaneously|
|Beckman Coulter MoFlo Astrios EQ||4||21||Sort 6 cell populations simultaneously|
Examples of Flow Cytometry Applications
Investigating the expression level of the target protein after tumor treatment
Human JIMT-1 breast cancer tumors were grown in mice treated with vehicle or test substance. At the study endpoint, tumors were isolated and tumor material was disrupted with gentleMACS C tubes containers and the Tumor Dissociation Kit (Miltenyi Biotec) before antibody staining and flow cytometry analysis.
Gating: Single cells were selected for live events via live/dead dye. Human tumor cells were identified as negative for mouse CD45 marker and investigated for target expression on the cell surface. Untreated tumor cells served as positive control expressing the target on 99% of cells. Whereas only 14% of the tumor cells from treated animals were stained positive for the target.
In conclusion, treatment of JIMT-1 breast cancer with the test agent reduced the number of target expression tumor cells to 14%.