Re: motor end plates (Hermina Borgerink)

Hi Karen,

I have a method for motor end plates that is almost as ancient as I am.  It was
given to me by a former boss, in hand-written form in 1962, so I cannot give
you any references other than that it is called Asbury Method for Motor End
Plates.  And it does work (at least it did then).

1. Tissue is fixed for 24 hours in cold 10% neutral buffered formalin at 4
degrees C.
2. Sections are cut at 8 - 10 microns in the cryostat and picked up on
albuminized slides.
3. Slides are dried horizontally in a 37degree oven for one hour.
4.  A stock solution of saturated Alpha Naphtyl Acetate is kept in equal
quantities of distilled water and acetone (may be stored at 4 degrees C for
later use).
5. Add 1 - 2 drops of this solution to 30 ml of cold barbital buffer in a
coplin jar.
6.  Add 15 - 20 mg of Naphthyl Diazo Blue B.  Stir quickly and use immediately.

7. Place slides in coplin jar and stain at 4 degrees C for 30 - 60 minutes or
until stained.
8. Wash 3 minutes in running water.
9. Dehydrate rapidly in alcolhol, clear through xylene and mount in permount.

Background - yellow
Motor end plates - greenish black to black


0.05 M barbital buffer pH 8.0 (for 200 ml):
2.06 gm sodium barbital
100 ml of distilled water
34 ml of 0.1 N HCl
Mix; then dilute to 200 ml with 66 ml distilledwater.
Store at 4 degrees C

Diazo Blue B, also known as Naphthanil or O Dianisidine Tetrazotized Fast Blue

Good luck!


Karen Larison wrote:

> Meg,
> Molecular Probes has fluorescent alpha-bungarotoxins that are used to
> stain motor end plates.  Their technical assistance number is (541)
> 465-8353.  Their WEB address is
> Karen
> University of Oregon
> >I have two questions:
> >
> >1) Can anyone tell me about a stain for motor end plates that works and is
> >not too difficult to do in large quantities?
> >
> >2) I am doing whole mounts of hamster ears and am having trouble getting
> >them to lay flat on the slide.  Any suggestions?
> >
> >Thanks,
> >Meg

Hermina Borgerink, BA, HT(ASCP)IHQ
Department of Pathology
Wake Forest University School of Medicine
Winston-Salem, N.C. 27157
PH (336) 716-1538
Fax (336) 716-1515

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