*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****
Hi Doug,
Aside from TDE and glycerol for ~aqueous RI range, there are
Sca/e; Hama etal, 2011
Benzyl alcohol-glycerol; Clendenon etal 2011
ClearT; Kuwajima etal 2013
the last 3 can be adjusted for final RI. I've used Sca/e with only fair results, but not yet tried the BA-Gly or ClearT. To-Pro-3 can withstand dehydration and solvents, I've used with methyl salicylate-benzyl benzoate clearing.
The retina is challenging, you will need to verify and correct for axial distortions.
Glen MacDonald
Core for Communication Research
Virginia Merrill Bloedel Hearing Research Center
Cellular Morphology Core
Center on Human Development and Disability
Box 357923
University of Washington
Seattle, WA 98195-7923 USA
(206) 616-4156
[log in to unmask]
On Jun 19, 2013, at 2:29 PM, "Cromey, Douglas W - (dcromey)" <[log in to unmask]> wrote:
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> I have a lab that is starting a project that will involve confocal imaging of whole mount retinas (small rodents). We were able to image to-pro3 labeled nuclei through several layers to a depth of approximately 100um using a 63x water immersion objective.
>
> Is there anyone that does something similar that would be willing to offer up some sample prep tips?
>
> I recognize that spherical aberration becomes a significant issue when imaging that deep. Any suggestions for possible clearing techniques (hopefully something not too complicated)?
>
> Doug
>
> ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^
> Douglas W. Cromey, M.S. - Associate Scientific Investigator
> Dept. of Cellular & Molecular Medicine, University of Arizona
> 1501 N. Campbell Ave, Tucson, AZ 85724-5044 USA
>
> office: AHSC 4212 email: [log in to unmask]
> voice: 520-626-2824 fax: 520-626-2097
>
> http://swehsc.pharmacy.arizona.edu/micro
> Home of: "Microscopy and Imaging Resources on the WWW"
|