We are using the confetti mouse and are trying to image on a Zeiss LSM 510 microscope. Does anyone know the appropriate settings to use to visualize all four colors?
Which lasers, which filters and how many cameras do you have in your system? And do you use the ZEN-software?
It is impossible to separate CFP, YFP, and GFP without spectral unmixing. This requires collection of reference spectra from samples expressing these colors separately and then mathematically separating the colors. You need to have a spectral microscope to collect the spectra with sufficient resolution (around 10 nm). Do you have a 510 meta? If so, the microscope can do this in principle but the sensitivity is low. The Zeiss LSM 710 or 780 are more likely to give you a satisfactory result without photobleaching. The Leica SP8 is also a good option but scans the spectrum one wavelength at a time so may be a bit slower.
Wow! Thanks. We have LSM 510 and LSM 710 scopes, both inverted. And we use Zen. I will check on lasers and filters.
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