Csaba, I can't answer the fluorescent question but for light microscopic IHC I would stain for the NeuN using peroxidase-DAB followed by a PAS for lipofuscin followed by Mayer's or Harris's haematoxylin.
Result is Brown, pink, blue Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 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 szi...@bio.u-szeged.hu Sent: Thursday, 2 June 2011 4:44 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Lipofuscin and counterstaining DAb IHC Dear Histonetters! I have human brain samples (age 70, HUntington disease) and my goal is to quantitate changes in the number of neurons/glia cells with nuclei staining. I use NeuN IHC and Höechst. My first question is: - Throughout the whole sample there are a lot of lipofuscin signal fading the NeuN signal, we are not able to make any microphtograph for cell counting. I tried Sudan black staining and Sigma autofluorescent reagent with the same result. Could you advise some tips and tricks to avoid this? An alternative solution could be DAB IHC for NeuN and counterstaining with nucleus staining. Second question is: - what type of counterstain do you recommend to be able to identify the IHC and normal nucleus staining in the same section? Thank you in advance, best regards Csaba ---------------------------------------------------------------- This message was sent using IMP, the Internet Messaging Program. _______________________________________________ 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 Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, The Childrens Hospital at Westmead 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
