[Histonet] ? why does my gallbladder control not stain for bile

[Lee & Peggy Wenk]

I think I can explain some of it, but I'd love to pathologist's input into
the exact type of "bile" in the wall of the gallbladder.

I'm assuming you are doing the Fouchet reaction, using trichloractic acid
and ferric chloride. This reaction take bilirubin (which is a brown-yellow
color) and oxidizes it back biliverdin (which is a green color).

When red blood cells die, they are broken down in the spleen and bone marrow
to heme and globin. The globin is further broken down to amino acids and are
recycled by the body into new proteins. The heme part is broken down into
iron and biliverdin. The iron is sent back to the bone marrow to make new
RBC's. This leave the biliverdin, which the macrophages turn into
water-insoluble unconjugated bilirubin. When this insoluble bilirubin
arrives in the liver, glucuronic acid is added, make it a water-soluble
conjugated bilirubin. The bilirubins are brown-yellow in color.

The bile that is found in gallbladder is a mixture of conjugated and
unconjugated bilirubin, with some biliverdin (and salts and cholesterol).

So when doing the Fouchet stain, the brown-yellow bilirubin that in trapped
in the liver due to cirrhosis or obstruction is converted back to green
biliverdin (with a van Gieson counterstain). Our control for the Fouchet
stain is a liver with bile duct obstruction. 

I've never tried this stain on a gall bladder. So, this is the part I'm not
sure about, and where I could use a pathologist's input - why is the
gallbladder wall green? I'm supposing that it is because it has absorbed
biliverdin, not the bilirubin. If that is the case, then the Fouchet stain
won't work, because there is no bilirubin to convert back to biliverdin.

I'd love to hear other explanations, theories, conjectures, etc.

Peggy A. Wenk, HTL(ASCP)SLS
William Beaumont Hospital
Royal Oak, MI 48073

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Kathy
Cormier
Sent: Friday, April 27, 2007 10:46 AM
To: histonet <@t> pathology.swmed.edu
Subject: [Histonet] ? why does my gallbladder control not stain for bile

Hello All,

Why does my gallbladder control not stain for bile?  ( the mouse livers do
not stain either, but that is not unexpected ) It is freshly cut, the
solutions are commercially bought and appropriately stored, the stain seems
very straightforward... Shouldn't this stain the bile in the gallbladder? I
can see it in the block... all nice dark green...Am I just missing the
obvious? Am I clueless? (Don't answer that!) I did this stain like, one
other time, in the early 90's and we had a nice piece of liver w/ bile for a
control. I have read Carson's and Bancroft's books, and they say the best
control is a piece of tissue w/ bile, so the gallbladder should work, right?
Help! (and thanks!)

Kathy
Div Comp Medicine
MIT
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