leiker <@t> buffalo.edu
Fri Nov 18 11:59:34 CST 2011
Do you have access to a confocal microscope or one that does Z-stack imaging? You can find out the thickness of your tissue sections that way (after you've mounted them on a slide of course).
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Amos Brooks
Sent: Friday, November 18, 2011 8:55 AM
To: histonet <@t> lists.utsouthwestern.edu; ssegal2 <@t> slu.edu
Subject: [Histonet] microtomes
If you are concerned about the thickness of the sections being accurate to the setting, I would suggest picking up a cheap micrometer from a hardware store. (OK perhaps not cheap as you will want a quality one, but the cost of these isn't terrible.) You can't really measure 5 microns (or whatever you cut at) well, so you will need to take 10-50 sections of a blank block. Remember to keep a constant rhythm that you would use on a normal block. Measure the thickness before and after then divide these thicknesses by the number of sections taken. Remember, also, to take the measurements at the same temperature that you are cutting the sections at.
If your difference isn't exactly right how far off is it? You would then know if you should use a higher or lower setting on the microtome.
On Thu, Nov 17, 2011 at 10:33 PM, <histonet-request <@t> lists.utsouthwestern.edu
> Message: 13
> Date: Thu, 17 Nov 2011 16:02:50 -0600
> From: Salomao Segal <ssegal2 <@t> slu.edu>
> Subject: [Histonet] microtomes
> To: histonet <@t> lists.utsouthwestern.edu
> <CACdnK8HuZek7dGaePt--kjzy219YnwgmEpiWq5DkoRPjtN8Aog <@t> mail.gmail.com
> Content-Type: text/plain; charset=ISO-8859-1
> The settings in a rotary microtome may indicate the thickness of
> sections but...
> how do you know that it indeed cuts at the indicated thickness,
> particularly if it is say an old device that you inherit from somebody
> else's lab junk?
> Is there a way of measuring the magnitude of advances after each rotation?
> Solomon Segal
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