Re: Immunohistology

From:Victoria Baker <vbaker60@yahoo.com>

<P> <BR> <P>  <B><I>Anila Syed <syedab@totalise.co.uk></I></B> wrote: <BR> <BLOCKQUOTE style="BORDER-LEFT: #1010ff 2px solid; MARGIN-LEFT: 5px; PADDING-LEFT: 5px"> <P>Hi all,<BR><BR>I am relatively new to immunohistology, but need to stain mouse liver for cysteine dioxygenase.<BR><BR>I would like to ask a few very basic questions please.<BR>If anyone has experience of using these techniques, I would be grateful of their assitance.<BR>First of all, what would be a good section thickness?</P> <P>I cut my sections at 4microns.  I also soak my mouse livers on ice for atleast an hour.<BR><BR>Secondly, is it best to use coated slides or not and if so, what is the best coating agent?</P> <P>I use positively charged slides.  Any major distributor has them.  They run about $35.00 a box.  I found that adding something to the waterbath left a background on the slides and the staining was distorted.  Maybe others have had better luck..If you have to go with coating their is an older procedure I once used called "Neoprene" and if you can get past the stench of it when coating the slides works very well.<BR><BR>Many thanks in advance for your help,<BR><BR>Anila Syed<BR><BR>Best of luck.  IHC isn't that hard to learn, but it can get frustrating at times.  Just change one "variable" at a time and don't try to rush something.  The internet has an excellent source of information procedures and many companies are very willing to help as well.</P> <P>Vikki Baker</P> <P>American Health Foundation</P> <P>Valhalla, New York</P></BLOCKQUOTE><p><br><hr size=1><b>Do You Yahoo!?</b><br> Send instant messages & get email alerts with <a href="http://im.yahoo.com/">Yahoo! Messenger</a>.
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