Hi Linda,
	First, get hold of an old portable sphygmomonometer [WHAT A GREAT
WORD!!!!  (did I spell it correctly?)] with a mercury manometer.  The
purpose is to enable yourself to track max pressure for perfusion.  The Hg
always falls as the perfusion continues and assures you that you are NOT
overpressurizing the system.
	Second, get hold of two used, glass, perfusion bottles (1L, with
hangers) and fit your own stoppers.  You will also need a ring stand with a
double burette clamp to hang the bottles on.
	Third, Tygon tubing, largest for elephant rats or caveys(sp???), but
nominally to fit over 5-8mm plastic tubing stubs stuck through the stoppers
(one long for air or pressure, and one short for outlet).
	Fourth, a three-way valve to select either of the bottles as outlet.
	Fifth, place a #20 hypodermic needle (blunted or not on a tuberculin
syringe barrel from which upper finger grip end has been removed (and
smoothed) and replaced with Tygon tubing from valve outlet.

	The procedure.
		1.  All solutions at 37C in the water bath until you are
ready to begin (but NOT to the second!!)
			a.  one=normal saline (Ringer's or buffer) flush (w/
or w/out ~1%(w/v) procaine (to relax vascular channels).
			b.  other= fixative or vascular casting
resin(surprise! - you need REAL pressure for that!)
		2.  For all posterior and anterior but heart (which requires
imaginative treatment) insert needle in left ventricle(?) and clamp
ventricle.  For lungs perfuse the other ventricle.  To concentrate on
posterior clamp or tie anterior arteries.  To concentrate on specific organ,
it is sometimes necessary to insert needle closer and to isolate organ from
adjacent systemic circulation (with substantial clamping but with care for
venous drainage).  Ligate around needle tip before perfusing with standard
heavy suture thread.
		3.  Set valve for saline flush and open appropriate venous
drainage with sharp scissors.
		4.  Add pressure slowly until blood begins to flow from the
venous drain.  Continue with MINIMUM pressure until drainage runs clear OR
until organ or area of interest shows signs of blood loss.
		5.  Switch valve to fixative and perfuse with minimum
pressure until fixation tremors are observed (sign that CNS is being
treated) or fixative flows from the venous drain.
		6.  Excise tissue and transfer to more fixative (cold) for
remainder of desired fixation period.

NOTE:  The sphygmo- thing is not required, but some pressure monitor is
preferred to insure that your setup is performing as it should.  There are
sphygmo's that don't use Hg as well.

	If you diagram this apparatus, you will realize that it is
essentially portable and can be assembled and removed at will.

Hope this helps.  

Fred Monson

Frederick C. Monson, PhD 
Center for Advanced Scientific Imaging(CASI) 
West Chester University of Pennsylvania 
Schmucker Science Center II  
South Church Street                                                    
West Chester, PA, 19383
eMail:  fmonson@wcupa.edu
http://darwin.wcupa.edu/casi/

  	

> ----------
> From: 	Linda Ryan
> Sent: 	Wednesday, October 31, 2001 11:38 AM
> To: 	HistoNet Server
> Subject: 	Setting up a perfusion lab
> 
> Hello,
> 
> I need to equip a lab from scratch for rodent perfusion.  The budget is
> limited.  Anyone using the ductless fume hoods?  I would appreciate any
> info
> from equipment(hoods, pumps, tools, anesthesia set-up, etc) to procedures.
> 
> Thanks,
> Linda Ryan, BS, HT(ASCP)HTL
> 
> HistoMax, Inc
> P.O. Box 3711
> Cary, NC 27519
> 
> 919-656-8620
> 
> 
>