Suitable fluorophore combinations for Tausense lifetime-based dye separation

Hello,

I wondered if anyone would be able to share fluorophore combinations that are appropriate for making the most of the Tausense module in multi-channel immunofluorescence experiments for imaging on a (new!) Leica Stellaris system. I’ve had a look for values of lifetime for common IF fluorophores (e.g. ISS Data Tables | Lifetime Data of Selected Fluorophores),
but since these are dependent on solvent etc and measured in vitro, how useful are they for work in tissue sections?
Can you, for example, stain Cy3 and AF555 together?

I’m obviously going to ask Leica for their suggestions on this, and see if they publish any materials to help design for Tausense, but I’d be grateful to hear from anyone with experience of using this.

Presumably this could look quite different from the way you would conventionally go about designing antibody panels for systems using spectral detector or unmixing-based methods of dye separation.
Or is it not sensible to use Tausense in this way at all - i.e. should you only use it for removing autofluorescent components, and try to keep a more conventional wavelength separation between your fluors (especially now with all that space out in near-IR on Stellaris :slight_smile: ?

I’d be grateful for any advice and examples. Thanks!

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