*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopyPost images on
http://www.imgur.com and include the link in your posting.
*****
Dear Tim,
There are several hundred references to the use of the counterstain DRAQ5 with Zebrafish embryos.
Although you may be familiar with far-red DRAQ5 (and real-time cell health / viability dye derivative DRAQ7) as being red-excited it can be usefully excited at shorter wavelengths. In fact it has a second peak of absorbance at ~599 nm.
I have data from an Opera user whose red laser had failed mid-screen and switched to the 561 laser and saw minimal change in the brightness of the DRAQ5-stained nuclei. I can share an image and legend for this if it helps.
As a further alternative, you might consider the related CyTRAK Orange which is preferentially excited at 515 nm but should excite well at 561 nm and is detected in a channel centred on 610 nm or with a LP filter. Usefully, it is not excited off the red line (633/647 nm) sources.
It is spectrally compatible with FITC, GFP and homologues.
Both DRAQ5 and CyTRAK Orange are live cell permeant and therefore suitable to live- or fixed-end point imaging (with the usual provisos of time, temp, conc, .. and tissue access limited by special structures).
The spectra and associated raw data files are found in the "key info'" tabs at the bottom of the respective product pages...
www.biostatus.com/draq5
www.biostatus.com/cytrak-orange
Please feel free to contact me off-list if you need more details or specific references.
However, one recent example in LS..
Pampaloni F, Ansari N, Girard P, Stelzer EH. Light sheet-based fluorescence microscopy (LSFM) reduces phototoxic effects and provides new means for the modern life sciences. InEuropean Conference on Biomedical Optics 2011 May 22 (p. 80860Y). Optical Society of America.
Kind regards,
Roy
Sent from my iPhone
Roy Edward
Biostatus Ltd.
roy(at)biostatus(dot)com
+44 (0)1509 558163
www.Biostatus.com
The home of DRAQ5 & DRAQ7
------------------------------
Hello all,
Does anyone have a favorite 561 nm-ex DNA dye for labeling tissue such as zebrafish embryos? I love To-Pro-3 for confocal DNA analysis; it works more reliably than exciting DAPI with a 405 nm laser and it scatters so much less in tissue. However this time I need to use a 561 nm laser.
Many thanks,
Tim
Timothy Feinstein, Ph.D.
Research Scientist
University of Pittsburgh Department of Developmental Biology
Locked
