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Date: | Tue, 26 Jul 2011 10:04:05 -0500 |
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Dear Pedro--
On 7/26/2011 9:26 AM, Pedro Almada wrote:
> If you are curious what it's for, it's for some of our users who image whole
> drosophila brains for structural information. Their common complaint is that
> half-way through their z-stack they loose image brightness until the last
> slices are barely visible. Clearing methods like Methyl salicilate aren't
> really adequate for neuronal structure I'm told, which leads me to consider
> this method.
This is a long shot, but is there any chance that they're over-staining
their tissue? If they're using high concentrations of their
fluorophores, they may be absorbing most of the light in the top half of
the tissue, leaving no photons for the bottom half. Cutting back the
concentration of secondary antibodies would be expected to improve
penetration, if this were the case.
Good luck!
Martin Wessendorf
--
Martin Wessendorf, Ph.D. office: (612) 626-0145
Assoc Prof, Dept Neuroscience lab: (612) 624-2991
University of Minnesota Preferred FAX: (612) 624-8118
6-145 Jackson Hall, 321 Church St. SE Dept Fax: (612) 626-5009
Minneapolis, MN 55455 e-mail: [log in to unmask]
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