[Histonet] Masson trichrome and H and E

[Hugh Luk]

Hi Emily,

My $0. 02 by saying it's a challenge for research groups, like ours, to keep quality stains that are not used everyday/week/month.  

I agree with your intuition.  Buy or make new eosin.

As for stocking eosin (or hematoxylin), I buy premade by a reputable vendor and always keep an eye on it getting weak or the dye adhering to the bottle.  Oxidation hurts H&E stains, so I never return used dye to the "New" bottle once I have used them (I get rather oblique requests too).  Alcohols are similar, as they always pull water out of the air, so my final dehydrating alcohols are always fresh.  Xylenes are less fussy.

For manual Masson trichrome, I use Newcomer Supply as I appreciate Marcia Walsh's humor, but most vendors are pretty good.  250 ml of aniline blue is $28.90.  Works great.  I don't know why your kit was $800, but I would guess it would be the phosphomolybdic/phoshotungstic acid.  Her Masson kit (blue) is rather high too at $294 for the 250 ml.

For me, aniline blue solution crashes first in the trichrome (other than the Weigerts).  Have you any desire to make solution from powder?  $27.10 for 25 g of certified dye?  Just wondering.  Shipping to my state adds $50-$100 to the price, so I think about stuff like this.

Hugh
Hawaii


> ------------------------------
> Date: Tue, 3 Mar 2015 07:38:30 -0500
> From: Emily Brown <talulahgosh <@t> gmail.com>
> Subject: [Histonet] Masson trichrome and H and E
> To: "histonet <@t> lists.utsouthwestern.edu"
> 	<histonet <@t> lists.utsouthwestern.edu>
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> 
> Hello!
> 
> I am doing Masson trichrome manually (not with a machine) and I just found
> out the kit is $800!! I was thinking of buying the ingredients separately,
> but why the hell is it so expensive???
> Also, the people I was going to borrow their reagents from said their
> aniline blue is not very good and I wanted to replace it.  I only need
> about 250ml, what brand do you guys prefer?
> l know I can google this, but I want to know what you guys like and what
> works best.  This is for mouse kidney paraffin sections, 4 to 5 microns.
> Another question, I did H and E and there is no eosin staining.  I think
> the reagents are pretty old, so I thought that might be a problem.  Also
> because my lab is cheap, they were reusing the xylenes and EtOH for both
> rehydrating and rehydrating.  I told my boss this is probably not a good
> idea as the end steps will have stain in them.  And I also think this is
> why it didn't work! The EtOH is also really old so who know is the 100% is
> actually even close to 100% any more.  I'm buying new reagents, but if you
> guys think anything else would help, let me know.
> 
> Also, shoutout to Ann, I know you're reading these!! Join the list!!
> 
> Emily
> 
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