As a colleague just pointed out, I think I suffered a
serious "senior moment" when writing the attached. 
Obviously (I hope) I meant to say that Flex is
isopropyl alcohol that we use as a substitute for
ethanol.  Please don't flog too hard.  It was a Monday
after all.
Roger
--- Roger Moretz  wrote:
> 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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