In my experiment, the mice were given subcutaneous injections of GFP stable human tumor cells. Mice were also intravenous injection into tail vein of qDot655 labeled B cells. When I want to use in vivo imaging with Carl zess LSM710, the technician told me that qDot can`t use two-photo.Excuse me, do you know whether qDot655 and GFP staining cells can be seen using two-photon? if not, can you recommend other dye which can be seen together with GFP? thank you very much!
They reply:
Thanks for contacting technical support of Thermo Fisher Scientific.
Not all dyes can be used for TPE.
Molecular Probes® fluorophores have been tried and tested in TPE applications as conjugates of antibodies, proteins, dextrans, and other biomolecules.
You could refer to the link below:
http://www.lifetechnologies.com/cn/zh/home/life-science/cell-analysis/cellular-imaging/super-resolution-microscopy/two-photon-microscopy.html
From our website, Qdot655 is OK.
For GFP, you could refer to the link below:
http://www.sciencedirect.com/science/article/pii/S0006349599770657
The dyes are excited at their two-photon absorption maxima: TMR at 850 nm, EGFP at 920 nm.
it depends if the qdot655 fits your emission filters. Qdots are suppose to be very good 2P probes since they have a high quantum yield and thereby are very bright.
You can check the emission with your filter in the spectraviewer offered by lifetech (see link below). I would guess you would be able to excite both fluorophores at the same wavelength and would try something between 850 nm and 950 nm.
https://www.lifetechnologies.com/ch/en/home/life-science/cell-analysis/labeling-chemistry/fluorescence-spectraviewer.html
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