Does anyone have experience using Invitrogen's Countess II FL to detect and count cells that have been immunolabeled with fluorescently-conjugated antibodies? Thanks!
I think you need to know the excitation/emission wavelength of your fluorescent compound and use the matching light cube on the countess. Then you should be able to dig out the menu and 'gate' on fluorescent cells. You might need to manually adjust the cut off on to determine fluorescent+ and fluorescent- cells. Hope this helps.
I'm with Tech Support for the Countess II. We've tried surface antigen detection, for instance (such as anti-CD antibodies) but with poor detection; the signal just isn't bright enough to get the sort of data you would get for expression using flow cytometry. Conjugated primaries likely will be too dim. But if the antigen is strongly expressed or highly abundant, then there is a good chance that the signal will be bright enough with secondary detection. An even better chance of detection would be possibly if amplified, such as with biotin-streptavidin or tyramide signal amplification. In any case, we can't recommend antibody detection since we don't have validated in-house data to support it. If you need further tech support, though, please contact us at 541-335-0353 or [email protected].
- Badges
- Science topic
- Similar topics
- Physical Sciences
- Physical Phenomena
- Luminescence
- Fluorescence