<P> <B><I>Anila Syed <firstname.lastname@example.org></I></B> wrote: <BR>
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<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>
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