staining genitalia

[Eric or Pat Metzler]

Rich,

Safranin O works well for the sclerotized parts in combination with 
Chlorazole Black E.  I put the Safranin powder in water, I'm told others 
put the powder in alcohol.  The Safranin does not dissolve in the water, 
but when in the liquid the Safranin powder releases the red die.  A lot 
of particulate matter from the Safranin stays in the solution.

I mix up several drops of the Safranin O solution.  I use it, and then 
it dries out in the air.  When I need it again I just add water to the 
dried residue.  Eventually the dried powder stops producing red die, and 
I make another few drops.  I mount the genitalia of a lot of 
microlepidopterans, and I just place a drop or two of Safranin O stain 
on a glass slide.  I keep an eye on the genitalia with my stereoscope so 
that I do not lose sight of them in the mix of stain and particulate matter.

Whereas the Chlorazole Black E can turn membranes far too black in a 
just a few seconds, it takes a bit longer for the Safranin O to turn 
everything red.  I rinse the stained parts in about 10% ETOH, then I 
transfer them to a series of 100% ETOH solutions for dehydration.  I 
flatten the parts white going through the 100% baths.

I hope this helps.

Eric

Richard Worth wrote:
> All,
> I was curious if anyone out there has done differential staining of 
> geni's and what stains they might use.  I'm very familiar with 
> Chlorazole Black E and its action on membranesbut  was interested in 
> something that is selective for sclerotized / chitonous structures.  
> They are sometimes faintly sclerotized and am looking for a 
> contrasting stain to the CBE to highlight those structures. Maybe 
> something pink, red, or orange.
> TIA,  Rich
>
>
> Richard Worth
>
> Entomologist / Lepidopterist
>
> Plant Division
>
> Oregon Dept. of Agriculture
>
> 635 Capitol St. NE
>
> Salem, OR  97301
>
> 503-986-6461
>
> rworth at oda.state.or.us
>
> http://www.oregon.gov/ODA/index.shtml
>
>
>

-- 
Eric H. or Patricia A. Metzler
P.O. Box 45
Alamogordo, New Mexico 88311-0045  USA
metzlere at msu.edu


 
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