RE: [Histonet] resin embedded tissue and sirius red staining

From:"Y. Wang"


We cut our sections at 6 microns although I think our microtome cuts 
thicker than it says. Someone from the list also recommended incubating at 
60 deg C before staining or using microwave heat. I'll try thinner 
sections and heat and let you know of the results (hopefully good).

Thank you both for your suggestions.

> I have done sirrus red staining on GMA sections with good results.  How
> thick are your sections.  I used 5 micron thick sections and stained with SR
> for 30 min.
> Patsy
> -----Original Message-----
> From:
> [] On Behalf Of Y. Wang
> Sent: Wednesday, June 01, 2005 2:29 PM
> To:
> Subject: [Histonet] resin embedded tissue and sirius red staining
> Dear histonetters,
> I have a question regarding the feasibility of staining resin embedded skin
> (in technovit) with Picosirius red (for collagen). We have some old samples
> embedded in resin that we would like to stain for collagen if at all
> possible. We normally do our sirius red staining on frozen and paraffin
> embedded tissue with good results. I tried running the same protocol (1hr
> sirius red followed by quick wash in acidified water) as well as at longer
> times (sirius red for 3 hours) with the resin sections but I get very dull
> staining (under polarized light I do not get the nice
> red/green/orange/yellow I normally see, instead it's rather murky). I
> haven't found anything online in terms of staining resin embedded samples
> with sirius red so I was wondering if this was one of those things that
> can't be done. Any comments would be much appreciated.
> Thank you
> Yak-Nam Wang
> Senior Fellow
> Department of Bioengineering
> University of Washington
> Box 357962
> Seattle, WA 98195
> Tel.: (206)-221-5873
> Fax.: (206)-221-5874
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