Bob:
Flex is an alcohol substitute, Clear Rite is a xylene
substitute.  We get both from Richard Allan.  Both
have lower vapor pressures than either ethanol or
xylene; therefore occupational exposure is reduced;
disposal or reuse of xylene is not an issue either. 
If you use ethanol and xylene, reduce the times I gave
you--probably by 25 to 33%--it will take some trials. 
I think the books reference by the other respondents
might give you some possible times.  The most critical
tissues are liver, kidney, brain and skin.  Our use of
the alcohol and xylene substitutes are as much related
to safety and disposal issues as anything else.  We
use large volumes of chemicals because of the number
of studies that are carried out in the environment.
Roger
--- r-meyer2@northwestern.edu wrote:
> Roger, 
> 
> I have never used Flex and Clearite 3 although I
> would be open to trying it.  
> Just wondering why you use those reagents, and do
> you think I could use your 
> processing schedule with ethanol and xylene?  We
> also use a VIP processor. 
> > 
> > Bob: 
> > We process animal tissue almost exclusively--mouse
> and 
> > rat being the two primary small animal species. 
> The 
> > primary thing is not to overprocess.  For mice
> (and 
> > maybe hamster), most processing steps are 30min,
> with 
> > two 1hour steps in paraffin (Shandon Hypercenter
> XP) 
> > or four 30min steps in a VIP E300.  (Of course,
> you 
> > could set up the VIP identical to the Shandon.)  I
> 
> > really like the Shandon for mouse tissue--cuts
> like 
> > butter, with no overprocessing issues.  With rat 
> > tissue, we go to a 45min step for dehydration and
> then 
> > into clearing for 1 hour.  Paraffin is two 1.5hr
> steps 
> > on the Shandons or four 45min steps on the VIP. 
> We do 
> > occasionally get some overprocessing of liver on
> the 
> > VIP, but brains and skins are generally
> underprocessed 
> > on the Shandon.  If I had the time I would try to 
> > refine the rat schedule a bit, but our workload
> rather 
> > mitigates against such things.  A couple of notes 
> > about our processing:  we use Richard Allan's Flex
> and 
> > ClearRite 3 for processing, not Ethanol and
> Xylene; on 
> > the VIP's, we use the "agitation" setting, but not
> the 
> > P/V on dehydration/clearing steps.  P/V is used
> only 
> > on the paraffin steps.  I have had some people
> play 
> > with the programs using P/V during dehydration,
> and 
> > was that tissue ever overdried and crunchy!! 
> Other 
> > specifics:  we use a 70, 80, 95, 95, 100, 100%
> Flex, 
> > 1:1 Flex:CR3 and 2 steps of CR3.  (we start with
> NBF). 
> > Hope this helps. 
> > Roger 
> > --- r-meyer2@northwestern.edu wrote: 
> > > Could anyone guide me to some good books on
> animal 
> > > processing.  We work mostly 
> > > with mouse, rat, and hamster tissues.  We get a 
> > > variety of organs as well so I 
> > > need a book or books that would cover how to
> process 
> > > different organs.  Also, 
> > > just wondering if there is a lab that has a lot
> of 
> > > experience in this area that 
> > > I could get in contact with for help in getting
> good 
> > > processing schedules going 
> > > for the variety of tissues we work with. 
> > > 
> > > Thanks 
> > > Bob Meyer, HTL 
> > > Northwestern University 
> > > 
> > > 
> > 
> > 
> > __________________________________________________
> 
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> > 
> 
> 


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