[Histonet] mouse perfusion rate

Fri Apr 2 14:02:08 CDT 2010

I wonder what pressure you were working at?  You can't tell if you
control flow rate.  It may have been very high.

Certainly, it would be possible to put the pressure high enough to
produce all the problems you describe, and fluid would come out the nose
as a part of that.  But how high is that?  

Is it possible to work at a pressure where fluid comes out the nose,
without blowing out the lungs or the liver?  I examine only the brain,
but a pressure that has fluid coming out the nose and yields excellent
brain perfusions is definitely possible (300 mm Hg for about ten seconds
or less, then lower pressure fixative flow).  And we don't notice
anything looking amiss in the rest of the body, which is also perfused.

Cordially,

Charles W. Scouten, Ph.D

Product Manager, MNL

Biosystems Division

Leica Biosystems Richmond, Inc.
5205 Route 12
P.O. Box 528
Richmond, IL 60071
United States of America

Telephone 630 964 0501

facsimile +1 630 964 0576

www.MyNeuroLab.com <http://www.myneurolab.com/> 

www.leica-microsystems.com <http://www.leica-microsystems.com/> 

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From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Merced M
Leiker <leiker <@t> buffalo.edu>
Sent: Thursday, April 01, 2010 8:03 AM
To: "Andrea T. Hooper" <andreahooper <@t> rocketmail.com>;
saby_joseph_a <@t> yahoo.com; making <@t> ufl.edu;
histonet <@t> lists.utsouthwestern.edu;
Charles.Scouten <@t> leica-microsystems.com
Subject: RE: [Histonet] mouse perfusion rate

Someone I believe on Histonet a couple years ago told me the same thing.
The "perfusion circuit" is compromised if you have fluid coming out the 
nose and internal organs swelling up... 

Merced 

--On Wednesday, March 31, 2010 4:14 PM -0700 "Andrea T. Hooper" 
< andreahooper <@t> rocketmail.com> wrote: 

> 
> Very interesting! Coming out the nose is definitely bad for any work I
> have done in the past - lungs get blown out, liver doesn't perfuse
well 
> and bone marrow looks horrific. However, if you are working with PFA
and 
> doing a post-fix anyway, you will probably be fine. If you are using
GA 
> and counting on the perfusion to ensure excellent fixation for things 
> such as lacZ staining (b/c post-fix in GA never works well for bone or
> deep into tissues) then blowing it out the nose is bad. Very bad. 
> 
> Andrea 
> 
> 
> --- On Mon, 3/29/10, Charles.Scouten <@t> leica-microsystems.com 
> < Charles.Scouten <@t> leica-microsystems.com> wrote: 
> 
> 
> From: Charles.Scouten <@t> leica-microsystems.com 
> < Charles.Scouten <@t> leica-microsystems.com> 
> Subject: RE: [Histonet] mouse perfusion rate 
> To: leiker <@t> buffalo.edu, saby_joseph_a <@t> yahoo.com, making <@t> ufl.edu, 
> histonet <@t> lists.utsouthwestern.edu 
> Date: Monday, March 29, 2010, 6:26 PM 
> 
> 
> I have perfused mice and rats at 300 mm Hg, about double physiological
> level, don't know what that made the flow rate. All mammals have the 
> same blood pressure (within tolerances), so it is easier to select a 
> suitable pressure to use than a flow rate, which varies dramatically. 
> 
> 
> 
> I look at brain, never pay any attention to the gut. Clear fluid comes
> out the nose, that is a good sign. There are pressure release valves 
> across the cribiform plate to release CSF if there is too much. I am 
> flooding the system, fluid coming out the nose means the extracellular
> fluid and CSF is being replaced as well as vascular blood. Good. 
> 
> 
> 
> The tissue is quality is excellent, free of red blood cells, can be 
> unshrunk depending on the tonicity (should be sub isotonic) of the 
> fixative fluid. Have looked at Nissl and EM material, no evidence of 
> damage to the tissue. 
> 
> 
> 
> If gut is extended, might have something to do with the large
intestines 
> job of removing fluid from feces, and flooding the system swells the 
> tissue. But does it matter? Do you use that tissue? What is the 
> tissue quality if you use it after physiological pressure perfusion. 
> 
> 
> 
> Cordially, 
> 
> Charles W. Scouten, Ph.D 
> 
> Product Manager, MNL 
> 
> Biosystems Division 
> 
> 
> 
> Leica Biosystems Richmond, Inc. 
> 5205 Route 12 
> P.O. Box 528 
> Richmond, IL 60071 
> United States of America 
> 
> Telephone 630 964 0501 
> 
> facsimile +1 630 964 0576 
> 
> www.MyNeuroLab.com < http://www.myneurolab.com/> 
> 
> www.leica-microsystems.com < http://www.leica-microsystems.com/> 
> 
> 
> 
> IMPORTANT - This email and any attachments may be confidential. Any 
> retransmissions, dissemination or other use of 
> 
> these materials by persons or entities other than the intended
recipient 
> is prohibited. If received in error, please contact 
> 
> us and delete all copies. Before opening or using attachments, check 
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> opinions expressed in this email are those of the 
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> individual sender]. 
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> 
> 
> 
> 
> From: Merced M Leiker < leiker <@t> buffalo.edu> [mailto:Merced M Leiker 
> < leiker <@t> buffalo.edu>] 
> Sent: Monday, March 29, 2010 9:05 AM 
> To: Joseph Saby < saby_joseph_a <@t> yahoo.com>; 
> Charles.Scouten <@t> leica-microsystems.com; making <@t> ufl.edu; 
> histonet <@t> lists.utsouthwestern.edu 
> Subject: Re: [Histonet] mouse perfusion rate 
> 
> 
> 
> Hi Joe, 
> 
> Thanks for that notice about flow rates. But I think for the mouse you
> meant 1-3mls/min (not per 10min?)... 
> 
> Regards, 
> Merced 
> 
> --On Saturday, March 27, 2010 5:03 PM -0700 Joseph Saby 
> < saby_joseph_a <@t> yahoo.com> wrote: 
> 
>> 
>> 
>> All- 
>> 
>> From previous work with rat perfusions, the flow rate was about 10 
>> ml/minute. If I had to guess, the equivalent flow rate for a mouse 
> would 
>> be closer to 1-3 mls/10 minutes. If you go 10 ml/minute, you will 
>> definitely cause blowout artefacts. 
>> 
>> Joe Saby, BA HT 
>> 
>> 
>> 
>> 
>> __________________________________________________ 
>> From: Merced M Leiker < leiker <@t> buffalo.edu> 
>> To: Charles.Scouten <@t> leica-microsystems.com; making <@t> ufl.edu; 
>> histonet <@t> lists.utsouthwestern.edu 
>> Sent: Fri, March 19, 2010 9:21:38 AM 
>> Subject: RE: [Histonet] mouse perfusion rate 
>> 
>> The vasculature will leak too much and the mouse will get bloated - 
>> you'll 
>> see it first in either the intestines blowing up like a balloon or 
> fluid 
>> coming out of the nose. Just not the same as the heart pumping when 
> the 
>> mouse is alive with intact physiology and normal functioning. Don't 
> know 
>> exactly why, but that's what happens when you go too fast. Perhaps
the 
>> vasculature has lost its control to compensate for the pressure? I'm 
> not 
>> a 
>> physiologist so I'm not sure why...maybe someone on the Histonet can 
>> answer 
>> that? 
>> 
>> Regards, 
>> Merced 
>> 
>> --On Thursday, March 18, 2010 5:49 PM -0500 
>> Charles.Scouten <@t> leica-microsystems.com wrote: 
>> 
>>> 
>>> 
>>> Why not? What happens? One would think the mammalian cardiovascular 
>>> system could withstand physiological pressures and flow rates, at 
> least 
>>> for one lifetime? 
>>> 
>>> 
>>> 
>>> 
>>> Cordially, 
>>> 
>>> Charles W. Scouten, Ph.D 
>>> 
>>> Product Manager, MNL 
>>> 
>>> Biosystems Division 
>>> 
>>> 
>>> 
>>> Leica Biosystems Richmond, Inc. 
>>> 5205 Route 12 
>>> P.O. Box 528 
>>> Richmond, IL 60071 
>>> United States of America 
>>> 
>>> Telephone 630 964 0501 
>>> 
>>> facsimile +1 630 964 0576 
>>> 
>>> www.MyNeuroLab.com 
>>> 
>>> www.leica-microsystems.com 
>>> 
>>> 
>>> 
>>> IMPORTANT - This email and any attachments may be confidential. Any 
>>> retransmissions, dissemination or other use of 
>>> 
>>> these materials by persons or entities other than the intended 
> recipient 
>>> is prohibited. If received in error, please contact 
>>> 
>>> us and delete all copies. Before opening or using attachments, check
> them 
>>> for viruses and defects. Our liability is limited 
>>> 
>>> to resupplying any affected attachments. [Any representations or 
> opinions 
>>> expressed in this email are those of the 
>>> 
>>> individual sender]. 
>>> 
>>> 
>>> 
>>> 
>>> 
>>> From: histonet-bounces <@t> lists.utsouthwestern.edu 
>>> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of 
> Merced M 
>>> Leiker < leiker <@t> buffalo.edu> 
>>> Sent: Thursday, March 18, 2010 12:38 PM 
>>> To: MKing < making <@t> ufl.edu>; histonet <@t> lists.utsouthwestern.edu 
>>> Subject: Re: [Histonet] mouse perfusion rate 
>>> 
>>> 
>>> 
>>> That may be mouse cardiac output, but I can assure you, from 
> experience, 
>>> you do not want to perfuse at 17ml/min. 
>>> 
>>> Regards, 
>>> Merced 
>>> 
>>> --On Thursday, March 18, 2010 1:32 PM -0400 MKing < making <@t> ufl.edu> 
>>> wrote: 
>>> 
>>>> Li, 
>>>> 
>>>> Mouse cardiac output seems to be about 17 ml/min (e.g. 
>>>> www.transonic.com/mice1.shtml), you probably want to try for that
to 
>>>> keep pressures close to physiological. 
>>>> A syringe pump is pretty inexpensive and probably all you need. 
>>>> 
>>>> Mike 
>>>> 
>>>> ----- Original Message ----- 
>>>> From: Li Zhang < dancingwing <@t> yahoo.com> 
>>>> Date: Wednesday, March 17, 2010 14:59 
>>>> Subject: [Histonet] question about mouse perfusion 
>>>> To: histonet <@t> lists.utsouthwestern.edu 
>>>> 
>>>> > > My question is: can anyone give me a rough idea of how fast I 
>>>> > > should inject ( like ml/min). I think I've tried like 30 ml in
3 
>>>> > > min, and I suspect that it's too fast because I do observe 
>>>> > > tissue swelling sometimes. 
>>>> 
>>>> 
>>>> 
>>>> _______________________________________________ 
>>>> Histonet mailing list 
>>>> Histonet <@t> lists.utsouthwestern.edu 
>>>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet 
>>>> 
>>> 
>>> 
>>> 
>>> Merced M Leiker 
>>> Research Technician III 
>>> Cardiovascular Medicine 
>>> 348 Biomedical Research Building 
>>> State University of New York at Buffalo 
>>> 3435 Main St, Buffalo, NY 14214 USA 
>>> leiker <@t> buffalo.edu 
>>> 716-829-6118 (Ph) 
>>> 716-829-2665 (Fx) 
>>> 
>>> No trees were harmed in the sending of this email. 
>>> However, many electrons were severely inconvenienced. 
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>>> 
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>> 
>> 
>> 
>> Merced M Leiker 
>> Research Technician III 
>> Cardiovascular Medicine 
>> 348 Biomedical Research Building 
>> State University of New York at Buffalo 
>> 3435 Main St, Buffalo, NY 14214 USA 
>> leiker <@t> buffalo.edu 
>> 716-829-6118 (Ph) 
>> 716-829-2665 (Fx) 
>> 
>> No trees were harmed in the sending of this email. 
>> However, many electrons were severely inconvenienced. 
>> 
>> 
>> _______________________________________________ 
>> Histonet mailing list 
>> Histonet <@t> lists.utsouthwestern.edu 
>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet 
>> 
>> 
> 
> 
> 
> Merced M Leiker 
> Research Technician III 
> Cardiovascular Medicine 
> 348 Biomedical Research Building 
> State University of New York at Buffalo 
> 3435 Main St, Buffalo, NY 14214 USA 
> leiker <@t> buffalo.edu 
> 716-829-6118 (Ph) 
> 716-829-2665 (Fx) 
> 
> No trees were harmed in the sending of this email. 
> However, many electrons were severely inconvenienced. 
> 
> 
> ______________________________________________________________________
> This email has been scanned by the MessageLabs Email Security System. 
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> 
> 

Merced M Leiker 
Research Technician III 
Cardiovascular Medicine 
348 Biomedical Research Building 
State University of New York at Buffalo 
3435 Main St, Buffalo, NY 14214 USA 
leiker <@t> buffalo.edu 
716-829-6118 (Ph) 
716-829-2665 (Fx) 

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