Re: Fw: a thank "u" + another ?

<< Previous Message | Next Message >>
From:Barry Rittman <> (by way of histonet)
To:histonet <>
Content-Type:text/plain; charset="us-ascii"

		depends on what you need to demonstrate. If you are
interested in fine
cytological detail then thin sections are probably needed. However it
should be remembered that if you use thin sections at either 2 or 4 microns
you are only seeing a portion of each cell. Even if you count nuclei your
count will be way high (is that good english?). Using a 5 micron thock
section,  a count of 100 cells may actuallly reflect a total number in 5
m,icron volume of tissue to be in the 30-40s.
(If you are interested in relationships between tissue components then
thick sections are ideal. I used to use EDTA separated mouse ear epithelium
which is around 30 microns thick to show Langerhans cells (to demonstrate
Iad and Iak antigens). The procedure worked extremely well and as we were
examining the entire epithelial thickness, we could obtain accurate numbers
of cells per unit area/volume. The limiting factor is the penetration of
the reagents,the time you are willing to spend on the reaction, your marker
and if you have the tissue fixed or not. If fixed can use overnight
incubvations if necessary. If unfixed, then there are some problems with
some tissues with diffusion of components and swelling of tissues rendering
evaluation difficult.
Even if you are using a thick section, if you use a fluorescent marker,
most of the time will examine using reflected ligh fluorescence microscopy.
This in effect is only looking at the top few microns of the section.

At 08:12 PM 2/18/99 -0500, you wrote:
>-----Original Message-----
>From: Jim Ball <>
>To: <>
>Date: Thursday, February 18, 1999 5:26 PM
>Subject: a thank "u" + another ?
>>Thanks to every one that responded to the thyroid question, but I had
>>already tried most of the solutions suggested. The one thought that was
>>along the lines I was thinking along were the S. bodies mentioned by one
>>person on the net. I did'nt check the dx. today but I will make apoint to
>>find out if they were present tommorow.
>>       My second mind grabing question is what do most of you feel is the
>>right tickness for IHC tissue section. I have read with interest the fact
>>that IHC seems to work well on thin sections. I have had some luck with 2
>>micron sections but every one seem to think 4 micron sections are the way
>>go. I really hate cutting thick sections and losing cytologic detail. Any
>>thoughs along this line. Oh yea my speeling stinks, so all English major
>>need not respond it does nothing for the tinctorial quality of my work.

<< Previous Message | Next Message >>