Wow these are awesome, thanks everybody for feedback! So appreciated Sent from my iPhone
> On Oct 2, 2014, at 7:05 PM, "Tony Henwood (SCHN)" > <tony.henw...@health.nsw.gov.au> wrote: > > The increased eosinophilia, in this scenario, is usually due to > over-decalcification. > > I would suggest: > > * Treat sections with 1% Periodic acid (as used in the PAS stain) for 10 > minutes before Haematoxylin staining. > * Use an iron-haematoxylin or Celestine blue-Haematoxylin > * decrease time in eosin (or use a neutral 1% eosin stain). > > This should increase nuclear staining and decrease the eosin reaction. > > Regards > Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) > Laboratory Manager & Senior 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: histonet-boun...@lists.utsouthwestern.edu > [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Wait, Trevor > Jordan > Sent: Friday, 3 October 2014 4:11 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Decalcified Bone Eosin Overstaining > > Hey histonetters! I'm currently doing some H&E Staining for EDTA Decalcified > Bone Tissue and it seems that the tissue is staining very intensely with > Eosin...perhaps too much. Was curious to see from some of the more > experienced histotechnicians out there which methods you guys have used in > the past to perhaps fix an issue like this. Here is my protocol that I have > used once I have sectioned the paraffin embedded tissue. I have omitted the 1 > second Acid Alcohol Dip in a few of my trials and it seemed that this method > worked great to keep the tissue from overstaining with Eosin. However, this > is an important step for differentiation of the Hematoxylin...so I'm a little > puzzled about what to do next. Perhaps wash with tap water again after the > Acid Alcohol? > > > 1. Xylene - 3 Minutes > > 2. Xylene 3- Minutes > > 3. Xylene - 3 Minutes > > 4. 100% EtOH - 3 Minutes > > 5. 100% EtOH - 3 Minutes > > 6. 95% EtOH - 3 Minutes > > 7. 70 % EtOH - 3 Minutes > > 8. Filtered Harris Hematoxylin (Commercial Purchase) - 6 Minutes > > 9. Wash with running Tap water > > 10. 1 second dip in 0.5% 12M HCL Acid Alcohol > > 11. Eosin Y Solution - 1 Minute > > 12. 70% EtOH - 2 Minutes > > 13. 70% EtOH - 2 Minutes > > 14. 95% EtOH - 2 Minutes > > 15. 95% EtOH - 2 Minutes > > 16. 100% EtOH - 2 Minutes > > 17. 100% EtOH - 2 Minutes > > 18. Xylene - 1 Minute > > 19. Xylene - 1 Minute > > 20. Xylene - 1 Minute > > > Trevor Jordan Wait > University of Texas Health Science Center, San Antonio Class of 2017 MD > Candidate Abilene Christian University Class of 2013 Graduate B.S. > Biochemistry _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ********************************************************************************* > This email and any files transmitted with it are confidential and intended > solely for the use of the individual or entity to whom they are addressed. If > you are not the intended recipient, please delete it and notify the sender. > > Views expressed in this message and any attachments are those of the > individual sender, and are not necessarily the views of The Sydney Children's > Hospitals Network. > > This note also confirms that this email message has been virus scanned and > although no computer viruses were detected, The Sydney Childrens Hospital's > Network accepts no liability for any consequential damage resulting from > email containing computer viruses. > ********************************************************************************* _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
