Re: Spirochete controls?

From:Lee & Peggy Wenk

Three options I can think of:

1. MAKE THEIR OWN

Take fresh unfixed lung (slightly edematous is better) or fresh placenta
(with most of the blood squeezed/blotted out), and cut it into 3-5 mm cubes.

Take the pieces down to microbiology. Have them make a fresh broth of liquid
culture media and non-pathogenic spirochete in a large test tube/centrifuge
tube. (Micro will know which culture media is best to use for the
spirochete.) Put tissue pieces into the culture media/spirochete mixture.
Cover.

Incubate in 37 degree C. oven overnight to 24 hours.

Add fixative, and allow to fix for 30 minutes. Then pour out fixative and
culture media, and add fresh fixative. (The first fixative is diluted by the
culture media, which is why a 2nd "dose" is needed. Also, the culture media
has "bugs" in it, so it is better to fix the microorganisms, before dumping
it down the sink or wherever you disposal of it.)

Allow to fix overnight, then place in cassettes, process and embed as usual.

There won't be any inflammatory reaction in the tissue, but there will be
micro-organisms in the tissue, and both spirochetes and tissues will have
been fixed and processed as that lab's regular tissues.

P.S. This technique also works for gram + and - bacteria, AFB and fungus.

2. PLACENTA:

In some parts of the country, there is an increase in syphilis. There is
also an increase in women who are pregnant being infected.

Those who are in these areas, please have your pathologists notify you when
they receive a placenta with syphilis.

Then process the ENTIRE placenta. You can put several LARGE pieces in each
cassette, and process all of them.

Then, embed some of the block, after cutting the pieces to a more manageable
size and embedding them into more cassettes. (In other words, if the tissue
is 1.5 cm x 1.5 cm by 3 mm, use a razor blade or scalpel at the embedding
station, and trim it into 5 mm x 5 mm pieces, and embed each of these pieces
into a separate cassette. By my calculations, you should be able to get 9
more cassettes from this one piece of tissue.)

This saves on number of cassettes going through the tissue processor. Also,
you can then embed the "cut down" smaller pieces into just the bottoms of
the cassettes. That way, you can save money by using the "bottoms only"
cassettes and/or not having to clean the metal tops. Also, you will have few
cassettes to store, if you don't embed them all at that time (see next
paragraph).

The rest of the processed cassettes that you don't want to embed right now,
place 8 in a slide box (yes, 8 will fit in 2 layers of 4 cassettes each),
pour more paraffin into the box to cover the cassettes. Add a paper label on
top of the hardening paraffin as to what the tissue control is. Then write
the same information on the lid of the box, and store in a cupboard, until
you need more. Don't put on the bottom shelf of the cupboard, if there is a
light under the cupboard. Can melt the paraffin (been there, done that),

Of course, then contact Histonet, and be willing to share with the rest of
us.

3. ANIMAL LEPTOSPIRA

Become friends with your local veterinary lab.

If they get any cases of Leptospira in - have them contact you. There are
different Leptospira that can infect rodents, dogs, pigs, cattle and
raccoons. (Weil's disease is a jaundice or meningitis that occurs in humans
who are exposed to the urine of these infected animals. In the US, I think
the most common source is from dogs.)

Once you get a kidney from an infected animal, fix, process, and proceed the
same as #2.

Remember to share with Histonet, of course.

Good luck to all.

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



----- Original Message -----
From: "Morken, Tim" 
To: "'Histonet'" 
Sent: Friday, November 15, 2002 9:54 AM
Subject: Spirochete controls?


> Does anyone know a good source for spirochete controls? (we at CDC have
our
> own but don't give them out for controls. I am asking for someone who
needs
> controls but can't find a source).
>
> Tim Morken
> CDC, Atlanta
>




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