[Histonet] mouse perfusion rate

Thu Apr 1 08:02:48 CDT 2010

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/>
>
>
>
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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)
>>>
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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.
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>>
>> _______________________________________________
>> 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.