You could try using a red blood cell lysis buffer.
Protocol provided by most manufacturers is for whole blood, although this can be adapted for tissue.
Would need to wash tissue thoroughly afterwards to prevent deterioration of tissue and wash away any haemoglobin.
A typical blood protocol would be
Dilute one volume of cell suspension with 5-10 volumes of lysis buffer (155 mM NH4Cl; 10 mM KHCO3 and 0.1 mM EDTA) and incubate for 5 minutes at room temperature
This has been done on small tissue chunks satisfactorily.
Regards
Leigh Silvester
From: Confocal Microscopy List [mailto:
[hidden email]] On Behalf Of Moreto Elias, Jemina
Sent: 16 June 2010 12:38
To:
[hidden email]
Subject: red corpuscles autofluorescence and nuclear endothelial cells staining
Dear all,
I have two technical questions and I was wondering if you could have the solution.
1- Do you know a way to avoid the presence of red blood corpuscle in tissues before performing immunofluorescence staining? Or otherwise, is there any way to avoid their autofluorescence?
2- We are trying to stain the nucleus of endothelial retina vessels. We have tried TX100 permeabilization, as well as HCl with DMSO without success... DAPI is staining all kind of nucleus in this tissue but not endothelial cell ones.
Thank you in advance
Jemina Moretó Elias, PhD.
Confocal Microscopy
Cancer Epigenetics and Biology Program (PEBC)
IDIBELL, Catalan Institute of Oncology (ICO)
Avinguda Gran Via de l'Hospitalet, 199-203
08907, L'Hospitalet de Llobregat (Barcelona)
Tel. +34 932607500 ext 3176
[hidden email]
www.pebc.cat
This message has been checked for viruses but the contents of an attachment
may still contain software viruses which could damage your computer system:
you are advised to perform your own checks. Email communications with the
University of Nottingham may be monitored as permitted by UK legislation.
Locked
