[Histonet] Problems with Orange G staining

From:"van Damme, Sarah"

Hi guys

Is this the correct address to send questions to? If not, my apologies,
but here it is if so:


I am using Dane's staining and failing to replicate some
previously-obtained results for  vaginal cells. The stain procedure is
Mayer's haematoxylin, then bluing in lithium carbonate, then phloxin B,
then Alcian blue, and last, orange G (colour index 16230). The vaginal
samples originally came up orange, and buccal cells stained more

However, during the last few weeks, we have been unable to obtain these
results: the vaginal cells are now mostly 'dusky pink', what I would
call a light blue-based red. The buccal cells are now more orange. After
trying a change to every variable I could think of, I managed to get the
buccal cells staining more as they did in the previous staining runs
(pink/magenta): however, the vaginal samples are not staining orange as
they were previously. It seems to be a problem with the Orange G not
working as it did before.

Has anyone had experience with this stain not working in a similar
situation, or does anyone have a suggestion? I am attempting to make
sure that every variable is the same as it was during the time frame of
the earlier results, right down to checking the source of the alcohol
used for fixing slides, trying different lots of the stain powders, and
the brand of the slides used.

Any help is much appreciated.

Sarah Van Damme
Institute of Environmental and Scientific Research
New Zealand

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