RE: automated IHC - Dab disposal
From: | Philip Oshel <peoshel@facstaff.wisc.edu> |
How about reducing the volume with a roto-vap to say, one liter, then
running the potassium permanganate/sulfuric acid reaction on that? I
wouldn't think the DAB is volatile enough to come off before the
water, but am I wrong?
Phil
>Hi Hazel,
>thanks for your reply to my query. Just a 2 things that need clarification.
>Firstly, when you say "working DAB" is this the amount of DAB in the waste
>disposal container or the amount of liquid dab as used per run-excluding all
>the water used for washing out the machine at the end of a clean cycle ? i.e
>a run of 48 slides would use 200ul each = 9600ul, but then would be diluted
>by water for the "wash cycle?" am I getting you confused?
>
>Secondly, what volume are you dealing with each day? I get about 10l per
>week, and I am concerned that I will end up with huge carboys of pot permang
>and sulphuric acid and gently bubbling solutions lying all over the place!
>This is getting to be quite a problem!!!
>
>Best regards
>Louise
>
>-----Original Message-----
>From: Horn, Hazel [mailto:HornHazelV@exchange.ach.uams.edu]
>Sent: 13 November 2000 09:07
>To: 'Louise Taylor'; histonet
>Subject: RE: automated IHC - Dab disposal
>
>
>> Louise,
>> This is the procedure approved by our state water department for disposal
>> of used DAB waste from our Dako autostainer. You might need to check
>> with your state water department before using this procedure.
> [Horn, Hazel]
> 1. Prepare the following stock solutions:
> 0.2 M Potassium Permanganate (31.6 g KMnO4/liter)
> 2.0 M Sulfuric Acid (112 ml concentrated acid/liter)
> 2. For each 10 ml of working DAB solution, add:
> 5 ml 0.2 M Potassium Permanganate
> 5 ml 2.0 M Sulfuric Acid
> 3. Allow the mixture to stand for at least 10 hours. This will
>allow it
> to become noncarcinogenic.
> 4. To decolorize the mixture, add powdered ascorbic acid until the
> color disappears.
> 5. Neutralize the decolorized mixture with sodium bicarbonate.
>Test
> the neutrality (pH meter, pH paper or dipsticks)
> 6. Discard by pouring down the drain with copious amounts of
>running
> water.
>
> If anyone has a simpler method we would sure love to have it. By
>the way, our water department will not allow us to use the procedure using
> bleach.
>
> I hope this helps!!!
> Hazel
>
>
>> I asked this question last week, but due to technical errors received only
>> one reply, so I'm asking it again. How do labs using the Dako autostainer
>> deal with their DAB waste? Have it collected by a waste company? If so -
>> what do they do with it?
>> Deal with it in-house? Pour it down the drain?
>>
>> Best regards
>> Louise Renton
>>
>>
>>
>> Research Laboratory
>> Department of Anatomical Pathology
>> South African Institute for Medical Research
>> Johannesburg
>> South Africa
>>
>>
>>
--
}}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{
Philip Oshel
Supervisor, AMFSC and BBPIC microscopy facilities
Department of Animal Sciences
University of Wisconsin
1675 Observatory Drive
Madison, WI 53706 - 1284
voice: (608) 263-4162
fax: (608) 262-5157 (dept. fax)
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