Photo-activatable far-red dye for Zebrafish embryos

[Anton Kamnev]

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Dear colleagues,

On our site we are trying to set up a method for tracking of various components in live zebrafish embryos. Although methods of labelling and injection works well so far we, as everyone would expect, run into troubles with light scattering when imaging deep into the sample (40 um and deeper).

Therefore, I was wondering if you could share some of experience in that area.

So far our guesses to improve the quality of the signal is:

1. Use probes which could be activated (scattering still will be there, however contrast should be better)
2. Shift to fluorescent tags in far-red spectrum.

I guess ideally what I’m looking for is a dye which:
1. Could be attached to proteins in vitro
2. Could be photo activated by common laser lines, e.g. 405 or 488 nm
3. Emits in far-red (>700 nm)
4. Not toxic for live embryos.
5. Not very expensive (as we only starting up large number of trials will be needed)

The microscopes we have available are Yokogawa CSU-X spinning disk, LSMs (Zeiss 710/880), and wide-field systems all equipped with galvo FRAP systems.

Thanks in advance for you advises!

Sincerely,
Anton.

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Anton Kamnev, PhD
Imaging Manager
Mechanochemical Cell Biology Building
Division of Biomedical Sciences
Warwick Medical School
The University of Warwick
Coventry, CV4 7AL UK

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