Work With Histology Data

12/17/09
Goal: Reveal microstructure information of size around 1 micrometer from a 1 mm slab of tissue.

• Using 1 mm slab, only the top 10-15 micrometers will be clear enough for analysis.

Suggested solutions:

• Cut into 10-15 micrometer tissue. Pretty easy to achieve with instrument.
• Two-photon microscopy (very expansive, only 1 group in Brown own it. Possible collaboration)
• todo: check about the most suitable fluorescent dye for brain tissue with Prof. Berson.

12/30/09 from David
talked to Paul Kulesa about microscopy stuff for brain tissue. He thinks that the requesting histology and imaging are not too tricky and can probably be outsourced. He believes, also, that the tissue can be "cleared" in a way that will let confocal (as opposed to 2-photon) imaging work. He mentioned a particular stain TUJ1 that labels axons and neurons (he thinks). He thought that with clearing, even wide-field imaging could work. He sent a pointer to a paper presented by Glen MacDonald at the Stowers meeting we attended (Trevor and I). It may have been about clearing or just a pointer to that group. He thought that that UW group might be willing to collaborate, since it would be quite easy for them to do the imaging and possibly the prep. They do human stuff, which Paul doesn't handle as much.

Paper reference from the Rubel lab that Glen MacDonald presented at the Stowers microscopy workshop.
Three-dimensional imaging of the intact mouse cochlea by fluorescent laser scanning confocal microscopy.
<http://www.ncbi.nlm.nih.gov/pubmed/18573326?itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_RVDocSum&ordinalpos=10>
MacDonald GH, Rubel EW. Hear Res. 2008 Sep;243(1-2):1-10. Epub 2008 Jun 6.