Re: Histogel

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From:amos brooks <>
To:Jeffrey S Crews <>
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<!doctype html public "-//w3c//dtd html 4.0 transitional//en"> <html> Hi, <br>    Jeffrey Crews said that when using histogel " <i>It doesn't really stain</i> <br><i>with H&E; there's just a little ghost of the Histogel surrounding the</i> <br><i>sample on the slide.</i>" <br>    I was wondering if anyone had done any Immunohistochemistry  or special staining (particularly silver stains) using this stuff. (I haven't tried it.) And if they had was there any background staining. We avoid using excess proteins in our float baths for this reason, and I assume the same concern for this product might apply. <br>Happy Cupid Day <br>Amos Brooks <p>Jeffrey S Crews wrote: <blockquote TYPE=CITE>Yep, we use it for processing islet cells that have been spun down in a <br>collagen gel. We get a little pellet that is mostly water and turns into <br>*nothing* when processed. But, the investigator wants to see an H&E of <br>the pellet. <br>        So, we put a vial of Histogel in a beaker of water and heat it up to <br>about 60C until it melts. Then we put a little puddle (about the size of <br>a nickel)  of Histogel on a plastic dish, and orient the pellet in the <br>puddle. If it sticks up above the top of the puddle we wait until the gel <br>thickens and then add more Histogel on top. Then we put the plastic dish <br>on an ice tray until the gel hardens. When it's hard we take a razor <br>blade and cut out a strip containing the sample, then process that in a <br>cassette between two biopsy pads. (It's easiest to pick up the strip on <br>the side of the blade.) <br>        The histogel shrinks dramatically when dehydrated and becomes a little <br>flat membrane. We just embed the whole thing. It doesn't really stain <br>with H&E; there's just a little ghost of the Histogel surrounding the <br>sample on the slide. <br>        I would guess that since you have samples of more substance, you may <br>need to adjust your processing schedule a bit. I hope this works for you; <br>with a little thought you should be able to work out your orientation so <br>that you see what you want after processing. Perhaps if you worked fast <br>you could do your orientation under a dissecting scope before the gel <br>hardens. <br>        I think that Richard Allen is selling Histogel now, unless it's changed <br>hands again. We could use something else, by the way; we could make up <br>agar or gelatin, but these days we don't have the time to make up <br>anything that we could buy instead. <p>Jeffrey Crews, HTL (ASCP) <br>Organogenesis, Inc. <p>On Wed, 09 Feb 2000 14:59:39 -0600 "Molinari, Betsy" <br><> writes: <br>>Hi Histonetters, <br>> Has anyone had any experience using Histogel? I have some very small <br>>mouse vessels That are very difficult to orientate after processing <br>>and <br>>I thought Histogel may be something to try. <br>>Thanks <br>>Betsy Molinari <br>>Texas Heart Institute <br>>713-794-6524 <br>> <p>________________________________________________________________ <br>YOU'RE PAYING TOO MUCH FOR THE INTERNET! <br>Juno now offers FREE Internet Access! <br>Try it today - there's no risk!  For your FREE software, visit: <br><a href=""></a>.</blockquote> </html>
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