[Histonet] Xylene Substitutes

[ewj]

We have a small lab at a university in Beijing where we do diagnostic 
histopath for swine diseases.
The lab is shared with graduate students who make histomorphologic 
measurements on tissues like gut and
muscle as part of their research.  We have a Sakura autostainer.  Last 
summer one student left all of the
caps off of all of the containers in the DRS autostainer over a hot 
summer weekend and
vaporized a very large amount of xylene and ethanol into the
room and out into the hallway which is on a floor shared with clerical 
staff one of whom was in early
stage pregnancy.  There was quite a furor ensuing, and no matter what 
the cause, the foreigner in the
building and his heavy use of the laboratory was to blame.  I have the 
support of the leadership/management
at the building and weathered the incident but it was not a comfortable 
situation.

Reducing the use of xylene is very attractive to us.

We have now been using Rene's hot water detergent method for some time.  
I have noticed that
when I do the process we get more reliable removal of the paraffin than 
if my staff does it.  Also one
of my workers has small sensitive hands.  My hands are toughened up from 
growing up on a
farm and working in the oil fields, and working with cattle and horses 
for quite a few years,
so the hot water doesnt bother me much,
but it is a problem for her and she is an excellent and valuable staff 
member
and really I should be concerned about scalding any one.

Rene' had suggested that I could get one of those restaurant coffee 
'vats' with a spigot on the bottom
to make and hold the hot water detergent solution, but those are not so 
easy to come by.  I have found
some old sterilizers which are essentially glorified pressure cookers 
with some automatic control that were
cast aside. These I have rigged up with pt100 thermocouples and PID 
thermocontrollers and use them to heat the water.
They have a drain port so I have also affixed them with heat tolerant 
solenoid valves for input and to drain
the slide container, and water level sensors, all of which are connected
to a computer driven data access module via relays so that process can 
be automated  and repeatable and
have tweakable cycles.  It's easier for me to procure such stuff in 
Beijing than it is to find t-shirts that fit me.

This also provides ready access to crisp hot water which is great for 
cleaning up paraffin
  from tissue block molds and tables and floors,  clearing plugged 
cleaning lines from the VIP5 (like when students fail
to drain the retort prior to retrieving their cassettes), and what not.
We dont have hot running tap water in the university lab.  (It's a 
luxury in homes as well but
much more common in recent years.)

I havent got around to converting the VIP5 to 2-propanol and mineral 
oil.  It's on a different
floor with good ventilation and I have enough trouble getting the 
students to read the sign
to drain the retort (in chinese), so I am not needing another level of 
complexity there
just yet.

I have been fiddling with the detergent solution and have found what 
does not work and also
found a reasonable result with an imported powdered dishwasher soap.   
We used a water
bath at first which is a real pain to use, is slow, and unpredictable.  
Predictable hot water
is important.

We find much fewer problems with lost sections particularly brain
with this method than with the xylene-alcohol series.  We do pig work 
primarily
so we run lots of brain sections looking for PRV and CSF and strep.

I am glad that Rene' continues to push his xylene-free concept.  It has 
helped me
immensely

Wayne Johnson
Enable Ag Tech Consulting
Beijing

On 3:59, Rene J Buesa wrote:
> The best solution to eliminate xylene is to use isopropyl alcohol mixed with mineral oil.
> Xylene can be eliminated from staining by dewaxing with 2% aq. sol. of dishwasher soap.
> Before coverslipping oven dry the stained sections and cover directly.
> To clean tissue processors use a 2% mixture of strong lab lab-ware detergent.
> Do all of the above and your lab will be xylene free.
> René J.
>
> From: Tom McNemar<TMcNemar <@t> lmhealth.org>
> To: 'Adrienne Anderson'<rennie1108 <@t> yahoo.com>; "histonet <@t> lists.utsouthwestern.edu"<histonet <@t> lists.utsouthwestern.edu>
> Sent: Wednesday, February 6, 2013 5:36 AM
> Subject: RE: [Histonet] Xylene Substitutes
>
> Not familiar with SubX but we have used Americlear for many years with good results.  You just can't beat Xylene for some things though and still keep a little around.
>
> Tom McNemar, HT(ASCP)
> Histology Co-ordinator
> Licking Memorial Health Systems
> (740) 348-4163
> (740) 348-4166
> tmcnemar <@t> lmhealth.org
> http://www.lmhealth.org/
>
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Adrienne Anderson
> Sent: Tuesday, February 05, 2013 4:58 PM
> To: histonet <@t> lists.utsouthwestern.edu
> Subject: [Histonet] Xylene Substitutes
>
> Hello all,
>
> My lab is looking into xylene substitutes, and I'd love some feedback on what other labs are using. We currently use SubX, but are there other items out there more economical?
>
> Thanks,
> Adrienne
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