Re: [Histonet] safranin o staining-rat cartilage-reg

From:Geoff McAuliffe

Routine formalin fixation will not maximize the preservation of 
cartilage proteoglycans. CPC or alcohol fixation will be better.
Before proceeding, consult the papers listed below by Shepard and Mitchell.

J. Histochem. Cytochem. 24(5):621-629, 1976.
J. Ultrastruc. Res. 54-451-460, 1976.
Histochemistry 70:107-114, 1981.
Anat. Rec. 187(4):463-476.
J. Elec. Micros. Tech. 5:17-44, 1987 by Thomopoulos et al.

Geoff wrote:

>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.
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Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583; fax: -4029

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