[Histonet] PSR

[bettina.hutz <@t> orionpharma.com]

ello,

we have some difficulties with PSR staining and so far no trouble = shooting
advice helped us finding the reason for our problem. We use PSR as routine
staining in CV reserch studies in order to = measure the quantitive amount
of connective tissue in heart slides. Therefore the staining has to be
proper with a big contrast between yellow and red = areas. In the beginning
all went well, but now the staining quality has = decreased, the connective
trissue is not as bright stained as for example one year = ago. I use the
same batch of picric acid and Pirco Siriusred and have not = changed
anything in the staining procedure.=20 Does anyone has any idea what can be
the reason? My staining instruction here:

PSR (Picro-Sirius Red) staining of paraffin slides
(modified by Bettina Hutz and Ritva Inkinen)
1.	Staining procedure
	Deparaffination
	3x xylene				each min. 3 minutes
	2x ethanol 100%				short
	ethanol 90%				short
	ethanol 80%				short
	ethanol 70%				short
	aqua dest.				Short
	Staining
	Weigert=B4s Hematoxylin		10 minutes
	Running tap water			10 minutes
	Rinse with aqua-dest.
	Picro-Sirius Red			30-60 minutes
	Dehydration & mounting
	3x ethanol 100%			short
	2x xylene	 			short
	Mount with xylene based medium
2.	Solutions
2.1.	Weigert=B4s Hematoxylin

2.1.1	Stock solution A
Hematoxylin (Natural Black 1, C.I. 75290)	10,0 g
Ethanol 95%					1000 ml
Produce app. 2 weeks before use

2.1.2.	Stock solution B
Ferric chloride, 29% aqueous		40 ml
Aqua-dest.					950 ml
HCl conc.					10 ml

2.1.3	Usage solution
Equal parts of solution A and solution B, use immediately.
2.2  Picro-Sirius Red

2.2.1	Stock solution
Sirius Red F3BA					5,0 g
Picric acid, saturated, aqueous		500 ml
Allow to stand for 48 hours before producing the usage solution.

Thanks for your help:)


Bettina Hutz


Bettina Hutz
Research Assistant
Department of Discovery Biology
Orion Corporation, ORION PHARMA
Orionintie 1, P.O.Box 65, FIN-02101 Espoo, Finland
Tel. +358 10 429 2607,  Fax +358 10 429 2924
Email: bettina.hutz <@t> orionpharma.com