Verhoeff's staining solution uses an iron mordasnt, which is an
oxidising agent, and iodine, another oxidising agent. It is very easy
to overoxidise the hematoxylin, particularly if a stock alcoholic
solution is used. I always used to preweigh 1 gram hematoxylin and keep
in test tubes, then add the required amount of ethanol at the time I
made the working solution. Problems I had had with poor staining then
stopped.
Have you considered switching to an iron resorcin dye type method? They
are progressive, much easier to control and work very consistently.
Their drawback is that making the solution is a nuisance and staining
often needs overnight. I liked Humberstone's variant (greeny blue black
elastic) as the solution is stable for several years and improves over
that time with shorter staining times. Others like Miller's (black
elatic). There are others as well.
Bryan Llewellyn
John Shelley wrote:
Hello Histonetters,
I have been asked to work on a project that will require me to do a
Verhoeff/Masson's stain. I have run some samples through the stain from a
protocol that I found on the archives and it is not working to my satisfaction.
I am not getting the nice elastin fibers to show black like they should. I have
looked at the slides just before going into 2% ferric chloride and it appears
to be dull or limited staining. I am using a kit from PolyScientific and not
sure if that might be my problem or not. My question is, are there another
people who are running this special stain combo and would you be willing to
share the procedure and also who you are buying your reagents from to
accomplish the staining. I appreciate any help with this request. Thanks!!!
Kind Regards!
John J Shelley
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