sure. but i guess that works best on thin sections...by the way, i am using 40 um brain sections.
2009-08-21 TF 发件人： Colleen Forster 发送时间： 2009-08-21 02:48:47 收件人： tifei 抄送： 主题： Re: [Histonet] anti-bromodeoxyuridine staining Using the antigen retrieval works well too. Colleen Forster U of MN TF wrote: > use 4N HCl, no trypsin! 20 min roomtemperature, works very > weeeeeeeeeellllllllllllllll > > > 2009-08-21 > > > > TF > > > > 发件人： Kitchell, Marianne > 发送时间： 2009-08-20 05:04:01 > 收件人： histonet@lists.utsouthwestern.edu > 抄送： > 主题： [Histonet] anti-bromodeoxyuridine staining > > I am having a problem with my anti-bromodeoxyuridine (BRDU) stain. I > have done this for many years on chicken tissues fixed in Notox. > Now I have rat tissues (Notox again) and sometimes I get everything > staining (all nuclei) and sometimes I get next to nothing to stain. I > have recently started cutting all my sections (paraffin) at 4 um. > Instead of 5um which may be a partial effect. > I am pretty sure that my problem is in the 2.0 N Hydrochloric acid (HCl) > step and/or the trypsin step. > It seems that if I use freshly prepared HCl, I get a better response > than if I have previously prepared and stored my 2.0 N HCl or buy 2.0 N > HCl prediluted. This in turn affects the amount of time required in the > subsequent trypsin step. Every time I get a new bottle of HCl I need to > retitrate the whole stain. Any help here so I am not redoing this into > infinity? I am starting to live the movie Groundhog Day. > I have never been able to perform this stain without an additional > trypsin step even though I have seen it done this way in the literature. > Thank you all in advance. > > > Marianne Kitchell > Novus International > St. Charles, Mo. > > CONFIDENTIALITY NOTE > The information contained in this message may be privileged and/or > confidential > and protected from disclosure. If the reader of this message is not the > intended recipient, or an employee or agent responsible for delivering this > message to the intended recipient, please be advised that any reading, review, > forwarding, dissemination, distribution or copying, of this communication or > any attachment(s) are strictly prohibited. If you have received this e-mail > in > error, please so notify the sender immediately. Also, please delete it and > all > attachments from any servers or other hard drives. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ------------------------------------------------------------------------ > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet >
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