Hi Tony, I have not been fixing them but will try the 1% acetic in ethanol and the methanol solutions, thank you for the suggestion!
Best, Vivian On Mon, Dec 5, 2016 at 2:52 PM, Tony Henwood (SCHN) < tony.henw...@health.nsw.gov.au> wrote: > Vivian, > > After cutting the frozen sections, how are you fixing them? > > If they are air-dried then they will tend to give a washed out nuclear > stain. > I would suggest immediate fixation of the sections in either methanol or > to make even a brighter H&E: 1% acetic acid in ethanol (1ml glacial acetic > acid + 99ml absolute ethanol), about 1 minute should suffice for both > fixations. > > Regards > Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) > Principal Scientist, the Children's Hospital at Westmead > Adjunct Fellow, School of Medicine, University of Western Sydney > Tel: 612 9845 3306 > Fax: 612 9845 3318 > Pathology Department > the children's hospital at westmead > Cnr Hawkesbury Road and Hainsworth Street, Westmead > Locked Bag 4001, Westmead NSW 2145, AUSTRALIA > > > -----Original Message----- > From: Vivian Hou via Histonet [mailto:histonet@lists.utsouthwestern.edu] > Sent: Tuesday, 6 December 2016 7:58 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Weak Hematoxylin Staining? > > Dear all, > > We are doing some immunofluorescent stains that we want to pair with H&E > slides. We are using fresh frozen sections for both (unfixed tissue, human > diseased coronary arteries) but I am seeing very weak hematoxylin staining > to the point where I have to dial back the eosin (3 min hematoxylin, 1 dip > eosin). > > > I am using Gills 2 (newly purchased), I know its not necessary to filter > it but we have been just in case using Whatman no. 5 filters. No > differentiation due to the weak staining right now. > > > I would appreciate any thoughts on why the staining might be so weak, we > do a great deal of processing and imaging on the arteries (about 12 hrs @ > 25C) prior to histo and I wonder if this has cause the tissue to decay? > > > Thank you for your time, > V > > -- > ------------------------------------------------------------ > ----------------------------- > V Hou > Bioengineering | Human Photonics Laboratory > e: ho...@uw.edu | c: 206-999-3708 > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > This message is intended for the addressee named and may contain > confidential information. If you are not the intended recipient, please > delete it and notify the sender. > > Views expressed in this message are those of the individual sender, and > are not necessarily the views of NSW Health or any of its entities. > > -- ----------------------------------------------------------------------------------------- V Hou Bioengineering | Human Photonics Laboratory e: ho...@uw.edu | c: 206-999-3708 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet