Verhoeff's Van Gieson Stain


Hello Histonetters. I had a question regarding collagen staining in VVG stain. I'm following a protocol where the slides are incubated for 30 minutes in Verhoeff's hematoxylin, differentiated in 2% ferric chloride, rinsed in hypo solution to remove iodine, then counterstained in Van Gieson's solution for 5 minutes. The results should show black elastic fibers and nuclei and red collage on a yellow background (other tissue elements). I do get those results, however the collagen stains a very faint red. The Van Gieson solution is made with 1ml 1% Acid fuchsin (1gm acid fuchsin in 100ml distilled water) and 45ml saturated picric acid, and allowed to stand overnight before staining. Now my question, is this the best way to make Van Gieson's solution, and if it is, should I just counterstain with it for longer than 5 minutes to get a brighter red? I know these might be trivial questions, but I've never done this stain but once recently and I don't have the time to troubleshoot sin
ce my PI wants the results for a paper ASAP. Thank you for your time.

Nidal E Muvarak
Associate Research Specialist
Department of Biomedical Engineering
University of Wisconsin-Madison
1550 Engineering Dr.; Rm. 2158
Madison, WI 53706-1609
Lab: (608) 265-8921; Office: (608) 265-4205; 
Home: (608) 256-7934; Cell: (608) 332-6068

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