We don’t have direct experience or customer feedback on CF dyes for 6-color imaging on LSM 800, but because this instrument has tunable excitation and linear unmixing, you should have a lot of flexibility in dye choice. We would also recommend contacting Zeiss support to see if they have advice for selecting dyes for multi-color experiments for the instrument.
The violet excited 405/430 dyes are generally less bright and should be used for more abundant targets. The 754 dye would also be recommended for more abundant targets. Even with optimal excitation, the GaAsp detectors on the Zeiss LSM 880 are expected to be less responsive to NIR wavelengths. The other dyes are very bright and would be better for less abundant targets to minimize cross-talk.
Below we’ve listed the dyes we’d usually recommend for the standard laser lines on the LSM 880.
405 nm laser line
CF®405S, Ex/Em 404/431 nm; recommended for abundant targets
458 nm laser line
CF®430, Ex/Em 426/498 nm; recommended for abundant targets
488 nm laser line
CF®488A, Ex/Em 490/515 nm; bright and photostable
Note: CF®488A and CF®514 must be separated by spectral imaging if used together
514 nm laser line
CF®514, Ex/Em 516/548 nm; dye for spectral imaging
Note: CF®488A and CF®514 must be separated by spectral imaging if used together
561 nm laser line
CF®568 Ex/Em 562/583 nm; very bright and photostable
Note: CF®568 and CF®594 must be separated by spectral imaging if used together
594 nm laser line
CF®594, Ex/Em: 593/614 nm; very bright and photostable
Note: CF®568 and CF®594 must be separated by spectral imaging if used together
633 nm laser line
CF®640R, Ex/Em 642/662 nm; CF®640 will be brighter with on-center excitation
CF®660R, Ex/Em 663/682 nm; CF®660R is off-center for this laser, but very bright and extremely photostable; less likely to have cross-talk with CF®594
Tunable 2P Laser 720-1020
CF®754, Ex/Em 745/786; recommended for abundant targets (LSM800 detectors are expected to be less sensitive for NIR dyes)
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