Re: [Histonet] Black pigment on Bronch Lavages

From:"Lee & Peggy Wenk"

I'm not certain that carbon birefringes when viewed with polarizing lenses. Formalin pigment will, as does silica and asbestos. But I don't think carbon does.
Carbon will resist "bleaching" with potassium permanganate and sulfuric acid, while this will bleach out melanin.
Try this:
2 slides of the patient tissue, on silane or poly-L-lysine, dried for a couple of hours at 60 degrees C. (This procedure tends to pull tissue off slides.) And 2 slides of melanin, also on the same type subbed slides and dried of same. Use another patient's case where there is carbon in lung tissue, prepare 2 slides of this the same way for your carbon control.
Deparaffinize all 6 slides to water. Keep 1 pt. slide and 1 control slide in water.
Place the other 1 pt. tissue and 1 each of the other controls (carbon lung and melanin) in a solution of 99.7 mL d. water + 0.3 mL sulfuric acid + 0.3 g potassium permanganate for about 1 hour at room temp.
Gently rinse in d. water.
Remove excess brown-colored potassium permanganate from tissue with 1% oxalic acid for 1-2 minutes.
Gently rinse in d. water.
Combine all 6 slides together. Stain with H&E.
The melanin will be gone from the treated melanin control slides, but will still be there on the untreated control slides.
The carbon control will remain positive in the treated and untreated.
Then look at the patient's tissue - treated and untreated.
That's all I can think of to do at a histology lab level.
An EM scope with metal analysis would work, as would micro-incineration analysis. But those involve specialized labs.
Let us know what happens.
Peggy A. Wenk, HTL(ASCP)SLS
William Beaumont Hospital
Royal Oak, MI 48073
----- Original Message -----
Sent: Thursday, October 30, 2003 4:02 PM
Subject: RE: [Histonet] Black pigment on Bronch Lavages

As our pathologist was explaining his problem to me while we looked at the slide in particular under his microscope, he said that Carbon was not the item in question as it does not become refractile when he polarizes/darkfield his microscope (to be honest he was whizzing objectives and filters so fast I could barely follow him!).  I know carbon is very common in lung, but to my eye as well, it is too fine and regular to resemble the stuff I usually see. 

-----Original Message-----
From: []
Sent: Wednesday, October 29, 2003 3:31 PM
Subject: Re: [Histonet] Black pigment on Bronch Lavages

dear kathy, how have you ruled out carbon, as this is so common. peter h. dohan, md

-----Original Message-----
Sent: Oct 29, 2003 1:30 PM
Subject: [Histonet] Black pigment on Bronch Lavages

One of our pathologists and myself have been trying to identify some black intracellular pigmentation in a bronch lavage.  We have ruled out carbon, and bleaching the section did not work, therefore is not melanin.  It is a very fine dark black pigment and appears quite uniform in shape and size.  Our pathologist is thinking that it is lead (the patient is a long time professional painter), but lead stains are negative.  My other thought is aluminum deposits but have not yet stained for this.
I am hoping someone on the "Net"  may have some idea of what this may be, and if there is a method for demonstrating it.
Thanks very much in advance!
Kathy Johnston
Tech II - Special Stains
Anatomic Pathology - FMC
Calgary Laboratory Services
1403-29 Street NW
Calgary AB, Canada T2N 2T9
403-290-4093 fax

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