Previously I did some surface marker expression (CD40, CD86) of RAW macrophages and DC2.4 cells using flow cytometry. For RAW cells, results were not satisfactory. Now, I am optimizing apoptosis assay with a pancreatic cancer cell.
Designing a panel in flow cytometry means selecting your fluorochrome-conjugated antibodies so that their signal won't overlap while taking account of your cytometer specification and allowing you to distinguish your important cell populations.
Panel desigining in the cytometry means selecting antobiodies so that their emission spectra dont overlap with other antibodies in that assay. Here, excitation can be same as the dectetor will only detect the emission of that fluorochrome conjugated antibody.
In addition, whether your cytometer has the laser to excite those antibodies.
Further, you can also check the expression of antigen and pair fluorochrome accordingly, for example if the marker is less expressed on cell you can use bright fluorochrome and if the marker is abudantly expressed you can pair it with dim fluorochrome.
- Badges
- Science topic