Re: [Histonet] Movat's stain/hematoxylin

From:John Kiernan

Welcome to Histonet! You have several questions.
Try again, asking just one with each email 

If you are "new in the world of histology" you
have been thrown in at the deep end with Movat's
pentachrome (even deeper with someone's modification).
Does your boss know anything about these methods?

There are plenty of simple (3 or 4 steps) ways to
stain collagen. You don't need a method such as
Movat's pentachrome (10 solutions and 14 steps) to 
do such a simple stainng job. 

You should persuade your boss to buy the lab one 
or more books about staining methods. Books in
this field cost about $100. How many slides stained 
by the Movat method would you get from a commercial 
lab for $100? Ten? Five? (Will someone tell us?)

The van Gieson stain shows collagen very well
using only two staining solutions. The submucosa
of the intestine always stands out very well. If you 
need to see very thin fibres, the related picro-sirius 
red method is just as easy to do. For multiple colors
(if that's a necessity) one of the one-step trichrome 
techniques will probably be adequate. All the
methods mentioned in this paragraph are suitable
for a beginner and can be completed in 15-20 minutes.

John Kiernan
London,  Canada.
Nora Berghoff wrote:
> Hello everyone!
> I am new to this list, so first I would like to say hi to everyone!! I
> have been reading the archives and find that they are a wonderful source
> of knowledge!
> I am also new in the world of histology - which is why I am asking for
> your help:
> I want to stain canine intestinal biopsies for collagen using Russell's
> modification of Movat's pentachrome stain. I have stained my first
> sections today and have encountered some problems:
> First, most of my muscle tissue is not really red, but rather
> orange/yellowish from the saffron, as it seems. But some areas of muscle
> have stained red.
> Fibrin does seem to be stained well (if what I have identified as
> fibrin is in fact fibrin...;))
> What can cause such irregular staining?  How long after cutting of the
> sections are they "stable" (meaning they don't lose any staining
> ability) on the slide?
> Could it be that my cut sections are old and "dried out"? I have had a
> histo lab embed and cut them, which is about five weeks I
> figured something may have happened to the sections in the meantime?
> Also, my elastic fibers and nuclei are not really black (I stained them
> according to the protocol for 15 minutes in hematoxylin sol.). I found
> it difficult to find the right hematoxylin (there are so many of them!),
> so maybe I have the wrong one after all...
> Does anyone know how long the individual staining solutions are stable
> after preparing them? Maybe my solutions are bad...
> So many questions and "maybe's" - I would really appreciate it very
> much if I could get some suggestions on any of them!
> Thank you so much in advance!!
> Nora Berghoff
> Research Assistant
> Gastrointestinal Laboratory
> Dept of Small Animal Clinical Sciences
> College of Veterinary Medicine
> Texas A&M University
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