[Histonet] safranin o staining-rat cartilage-reg

From:PKamalavenkatesh@wockhardtin.com


Dear histonetters,

This is with regard to safranin o staining of proteoglycans of immature rat
cartilage.  I have followed the protocol from Gayle Callis. Unfortunately I
couldn’t  get  any  type  of  red  color  staining  of proteoglycans in the
articular  cartilage. I am having doubt if all the proteoglycans got washed
away.  The  joints  are  fixed  in  10%  neutral buffered formalin and then
decalcified  using  10 % EDTA solution for a week. Can anybody experiencing
the  same  problem  like  this.  In  the literature, t was recommended that
addition  of  cetylpyridinium chloride in the fixative will reduce the loss
of proteoglycans during fixation and processing. If anybody having any sort
of  experience  regarding  this.  Also can anybody provide me the Toluidine
blue staining methodology for cartilage.

Regards
REGARDS
Dr.P.Kamalavenkatesh
Pre clinical safety assessment division
New Drug Discovery- Biology
Wockhardt Research Center
India

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