Hi,
    I pulled this off the archives. I thought I remembered it but wasn't sure
where. Archives save the day again. I don't know if this would stain yeast cells
though. All I can say is give it a try and find out. By the way thanks go to
Kathy Paton for the method. I certainly can't claim credit except for having a
memory for weird details ;-)
Amos Brooks

***********************************************************

Dear Deborah     <------- (that's who Kathy Paton was responding to AAB)
I have used the following method with excellent results.
Bergs Stain for spermatozoa
SOLUTIONS
Putts carbol-fuchsin
   New fuchsin            1gm
   Phenol                 5gm
   Distilled water       84ml
Absolute alcohol         10ml

Saturated Lithium Carbonate
   Lithium Carbonate      1gm
   Distilled water      100ml

Glacial acetic alcohol
   Glacial acetic acid    5ml
   Absolute alcohol      95ml

Methylene Blue
   Methylene blue        0.5gm
   Absolute ethanol      100ml

METHOD

1   Sections to water
2   Stain for 3min in carbol fuchsin (filtered)   3 min.
3   Place in lithium carbonate for  3min
4   Decolourize in glacial acetic alcohol for 5min (control microscopically)
5   Wash in two 1-minute changes of abs alc
6    Counterstain in methylene blue for 30 to 60 seconds.
7   Rinse rapidly in a couple of changes of abs alc
8   Clear in several changes of Xylol...mount.

RESULTS
Spermatozoa   brilliant red
Erythrocytes  pale pink

Postive control is required.......this in itself can prove to be fun even in
a busy lab.
I will comment no more

Kathy Paton
Surgical Pathology Unit
North Shore Hospital
Auckland
New Zealand

m.brinkenberg@nfi.minjus.nl wrote:

> To all,
>
> The problem we have in the forensic field is that nothing is constant, we
> work with stains from clothes and slides made from the vagina ect.
> These stains are not fresh(mostly more than 3 days old) and therefore very
> vonurable for extern invirement.
> Some even have alot of yeast cells(like from the anus), and if sperm loses
> there tail, it is some times difficult to differntiate between spermcells
> and yeastcells.
> It is very important to diagnose if there is sperm or not(in a court of law)
> On the other hand the stain canot take to long, since we have alot of these
> cases(daily) and we can't spend much time on a case.
> We are testing acridine orange stains at the moment but I can't get
> satisfactorry results, I searched for relevant articels and book in the
> forensic field but can't find nothing.
> Hoping you can give me an advise,
>
> Sincerly Yours,
>
> Brinkenberg MNA