Mayer's hemaotoxylin (a "progressive" hematoxylin) is not adequate for FS. Try Harris hematoxylin BUT stain first with the hematoxylin and with ORO after wards to obtain better results. René J.
From: Stacey Barrick <barricksta...@yahoo.com> To: "Histonet@lists.utsouthwestern.edu" <Histonet@lists.utsouthwestern.edu> Sent: Wednesday, February 27, 2013 11:20 AM Subject: [Histonet] Mayer's Hematoxylin on frozen tissue Hi everyone, We are trying to stain frozen cut sections of aorta with Mayer's Hematoxylin following Oil Red O staining. We cannot get hematoxylin staining to work. We are mainly seeing blue background but not labeling of nuclei. Tissue is fixed with 4% PFA prior to sectioning. After tissue is cut; sections are stained with Oil Red O · ddH2O 2’ · 60% isopropanol 30s · Oil Red O 18’ · 60% isopropanol 30s · 2x ddH2O 1’ Sections are then stained with Sigma Mayer's Hematoxylin · Rinse in deionized water · Stain in Mayer's Hematoxylin 1-5 min · Rinse in running tap water until nuclei are blue · Rinse in deionized water We have tried staining in Hematoxylin for 3 min up to 15 mins We have also tried rinsing 1 min up to 15 mins (checking at for staining at various timepoints) and always see the same result -some blue staining but no clear nuclear staining. Does anyone have any suggestions as to why this isn't working? Thanks!! Stacey _______________________________________________ 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
