[Histonet] Masson trichrome and H and E

[John Kiernan]

If you can't get two colours with H&E, don't expect to get the colour scheme right with Masson's trichrome, which needs more skill. If you are hoping to show basement membranes in the kidney, you would do better to use a technically simpler staining method such as picro-sirius red or periodic acid-Schiff. If for some reason you really need three colours, a one-step trichrome such as Gomori's, Cason's or Gabe's might be the way to go rather than Masson's or one of the other multi-step trichromes. Remember that all trichrome methods are greatly influenced by the fixative. A post-fixation treatment of the sections, usually with picric acid, is needed for formaldehyde-fixed tissues. Some alternative post-fixation treatments were proposed by Yu & Chapman (2003) J. Histotechnol. 26(2): 131-134, but their coloured photos were not very convincing.

Making up staining solutions in-house is always cheaper than buying pre-made solutions.

John Kiernan
London, Canada 
= = =
On 03/03/15, Emily Brown  <talulahgosh <@t> gmail.com> wrote:
> 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
> 
> 
> "By bitching and bitching and bitching, they could exhaust the drama of
> their own horror stories. Grow bored. Only then could they accept a new
> story for their lives. Move forward."
> 
> -Chuck Palahniuk, "Haunted"
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