Author Topic: Picrosirius Red stain on cryosections  (Read 897 times)

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Offline CJurkiewicz

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Picrosirius Red stain on cryosections
« on: February 05, 2016, 10:55:53 AM »
Hi All

So I am staining for collagen using Picrosirius Red on some cryosections of rainbow trout ventricles.  I have done this before using paraffin sectioning before with no problem before.  When looking at the sections under the microscope the myocytes are also stained as well which is interfering with the collagen fibers.  I have done three different pre-treatments for these cryosections all to no avail.  The three methods I used were
1. Xylene: 2 x 3 minutes
2. Xylene 1:1 with 100% ethanol: 3 minutes
3. 100% ethanol: 2 x 3 minutes
4. 95% ethanol: 3 minutes
5. 70 % ethanol: 3 minutes
6. 50 % ethanol: 3 minutes
7. Running cold tap water to rinse

another method was just rinsing the slides with a 10% PBS solution multiple times and finally placing the slide in 10% NBF solution.  All three pre treatments yielded similar results with the myocytes interfering with the stain.  I was planning on looking at the compact layer of the ventricle in the rainbow trout heart and to compare it across some treatments that I did (if anyone was wondering)

thanks

(I can post a picture of the slides later once I get some more time on the microscope to better explain my situation)

Craig J   

Picrosirius Red stain on cryosections
« on: February 05, 2016, 10:55:53 AM »

Offline gula

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Re: Picrosirius Red stain on cryosections
« Reply #1 on: February 09, 2016, 02:41:46 PM »
xylen is only needed for deparaffination.
I would try to fix the slides directly in Bouin (30 min) after short air-drying. then rinse with tapwater until the sections are not yellow any more. then rinse in deion. water followed by the picrosirius-red procedure. then rinse in 96% ethanol, dehydrate, clear in xylene and coverslip.
gula

Offline CJurkiewicz

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Re: Picrosirius Red stain on cryosections
« Reply #2 on: February 09, 2016, 06:54:36 PM »
Thank you so much, I will let you know how it turns out


-Craig J

Re: Picrosirius Red stain on cryosections
« Reply #2 on: February 09, 2016, 06:54:36 PM »