Re: Gram Stain problem

From:Connie McManus <>


I'm having the same problem with my gram stain.  I'm using Brown and
Hopps. All the bacteria stain blue as if the differention never
happened.  I've been experimenting with Rosanaline vs pararosaniline to
see if that makes a difference, but so far nothing has changed.  I tried
Brown and Brenn, using both versions of Basic Fuchsin, and that didn't
work, either.  

Aidan, what are you using to differientiate with (I'm not familiar with
the Bancroft & Stevens method)?  I'm using pure Acetone.  I tried the
old microbiologist's standby of Abs ETOH:Abs acetone, 1:1 and that made 
no difference either.  

Sorry, but I'm as baffled as you!  

Connie M.

Aidan Schurr wrote:
> Hi all,
> We have been having troubles with our gram stain (paraffin) procedure.  The results are not giving sufficient distinction between Gram +ve's and -ve's.  It has worked in the past, and nothing should have changed.  I have tried numerous variations, new reagents, different differentiation times, and different people doing the stain, all with no luck.  What is the crucial part of this stain?  Where are we going wrong?!  I've been using the protocols from Bancroft and Stevens.
> Many thanks in advance for your help,
> Aidan, New Zealand.
> _____________________________________
> a i d a n   c   s c h u r r
>  section head, histology
>   department of pathology
> hutt valley health
> high street, lower hutt
> wellington
> new zealand
> telephone ++64 4 5709173
> facsimile ++64 4 5709214


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