[Histonet] yeast and bacteria on slides

[JOHN PHILLIPS]

May I suggest you consider this for starters.

The most likely cause is your water floatation bath. How many are used? Do you meticulously clean each one at the end of each day and fill up with fresh deionised water? Can you identify the water bath or baths(if several were used). Here, operator marks their slides with initials for audit purposes so we know who picked up the section, labelled the slide and so which water bath they were using. Hey, bloody audit coming in handy!! Water baths filled with water and left on for several days prior to use will grow micro-organisms, especially if they have been used and not cleaned. If you can pin point sections from each bath and have spare unstained sections from  each bath from the "day of recogning", then i suggest you stain a couple of spares from each with PAS and you may get your answer. It has happened here a couple of times in the past.

Hope this helps.

John, Wrexham, Wales.

-----Original Message-----
From: Melissa Jans [mailto:histomjans <@t> yahoo.com]
Sent: 01 February 2005 01:40
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] yeast and bacteria on slides


I need some help!!
We have been experiencing some yeast and bacteria showing up on some of our special stains.  It started about 2 weeks ago and seems to be sporadic (one day...no problems, the next day, several stains are affected).  The yeast and bacteria (rods) are clumped together all over the slide, on the tissue sections, around the sections and on the slide in areas where there is no tissue.  The stains we have seen the contamination are the Gram, PAS/D, PAS and GMS.  All of these stains are automated except the Gram, so I do not think the special stainer is the problem.
Here is what I have eliminated as problem areas:
-Processor solutions and paraffin (we cut and stained a block from another institute and had contamination) 
-Waterbaths, slide drying boards, water containers (used to fill waterbathes), stainer containers, ice pans (all of the mentioned have been thoroughly bleached)
-Tap water and DI water (all water supplies have been tested by sending a sample to micro...there was no bacteria or yeast detected)
-Stay-on used as a slide adhesive (smeared some on a slide and stained it)
-Bad batch of slides (we have had contamination on two different slide sources (vendors))
 
Does anybody have any ideas?  What am I missing?  Any suggestions would be much appreciated.
 
Melissa Jans
University of Iowa Hospitals and Clinics

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