Re: tissue embedding for microtomy

On Sun, 04 Mar 2007 09:08:07 -0500, Kevin Cunningham <smskjv@xxxxxxxxxxxxxx> wrote:



One question is how thick will the specimen be? In the past neuro people
have used quite thick specimens usually embedded in Epon or Epon like
materials. This was to allow the investigator to see the neural process.
Usually the thickness was about 100 to 125 microns. Now this is quite
thick, but neuro folks liked this process. You could reduce this to 10
microns as long as the process were straight.

Let us know!

Thanks,

Kevin Cunningham
SMS



Hi, and thanks for the response! I have a simple sliding microtome which claims to be able to cut .5 microns.I'm not sure if this tool can cut plastic materials which is why I'm thinking parrafin. I'm not really sure how thick to go but I figured I could get there experimentally based on what I could successfully cut and observe. I'm really trying to get a handle on how to process and embed specimens. The simplest and safest process I've found so far involves progessively purer grades of ethyl and isopropyl alcohol followed by alcohols blended with mineral oil and finally infiltrating and embedding in paraffin.I was thinking paraplast+ with dmso. The process involves no xylene, which I like. One pressing question I have concerns anhydrous alcohol: do I absoloutely have to have it or could I get by with 190 proof, at least in the beginning. I'm really talking about ethanol because I can get technical grade 99.5% isopropyl as cleaning fluid inexpensively and locally. If I could use 190 proof the local package store sells grain alcohol at that strength. Im picking nits here because I know how sensitive all this stuff is and I dont want to doom myself to failure at the start. Thank You,Harry.

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