[Histonet] Safranine o staining and Problem in tissues

[Connie McManus]

What are you looking for in the joint staining you want?  If you want
something with safranine O, Movat's Pentachrome comes to mind. Not
having worked with bone histology for over 20 years, I'm not the best
resource for this.  I do remember Movat's being a big favorite during
the time I worked specifically in bone histology.  It was done on
decalcified bone as well a MMA embedded bones. 

DECALCIFIER:  
Decal-Stat   from the Decal Corp.  PO Box 236, Congers, NY  10920;
Phone:  800/428-5856;  fax:  845/268-383
This is a rapid decalcifier.  We use it on all our tissues regardless of
anmal.  It works great.


As for your liver, kidney and brain sections, --- I'm going out on a
limb here --- but it sounds like it could be heat artifact.  Are you
putting your sections on a heating tray before staining them?  If so,
check the temperature... it may be way too hot.  My water bath is set at
37 to 38 C and I keep my heating tray set to 40 C.  That's pretty warm,
but it works well for tissues like gut, heart, skin, ovary, uterus
kidney, liver.  I don't leave any tissue on this tray for more than 10
or 15 minutes and haven't had any problems like this.  Generally, I
don't put brain, or fetal tissues, anything frail on the tray except if
they seem to need a little relaxing of the tissue.  In that case, I
watch the sections carefully and remove them as soon as they smooth out.


For what it's worth...
Connie McManus
Utah Veterinary Diagnostics Laboratory
Utah State University
Logan, UT
Phone:  435/797-1891
fax: 435/797-2805


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
balajimr <@t> drreddys.com
Sent: Tuesday, April 13, 2004 9:15 PM
To: histonet <@t> lists.utsouthwestern.edu
Cc: histonet-bounces <@t> lists.utsouthwestern.edu
Subject: [Histonet] Safranine o staining and Problem in tissues
Importance: High

Can anyone give me the detailed procedure for safranine o staining for
the 
joints (ankle. tarsal and metatarsal joints in rats). I would like to
know 
a good  decalcifying solution for the same and what is the duration of 
bone to be kept in the decalcifier. 


Secondly, of late I am facing  peculiar problem now with my liver,
kidney 
and brain sections of late. These sections are cracking after the
staining 
(Mayer's H& E). It is something like parched earth appearance. This I am

noticing when the tissue s are coming to last xylenes. I donot see the 
cracking before this step. This doesn't happen in all the slides. Could 
you please suggest some solution for this. We are using chloroform for 
clearing and histoplast from  Shandon for infiltration. I am unable to 
locate where exactly is the problem. I have tried changing the makes of 
chloroform and xylenes used with out much help. 



Dr. Balaji  (M. V. Sc. Path)
Dept. Pre clinical safety evaluation,
Discovery research, 
Dr. Reddys Laboratories ltd. Bollaram Road, 
Miyapur, Hyderabad, 500 049
 Andhra Pradseh, INDIA

Tel: 23045439 - Ext.432.
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