fluorescent stain for glass

[Martin Wessendorf-2]

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Are aerosols from cryostat-sectioned tissue an issue?  I know that when
I cryosection rat brains, the particulates are sufficient to awaken my
rat allergy and set me sneezing.  I would guess that with particulates
containing viruses or prions could also be inhaled.

Martin Wessendorf

On 8/31/2011 11:21 AM, Danielle Crippen wrote:

> The need to cryosection most surely means the tissue won't be fixed...maybe they will need ORO or Betagal staining or something along those lines that requires fresh frozen tissue.
>
> As long as the tissue isn't positive for HIV (or some other virus)which might require additional guidelines, I would approach this situation in the following manner:
>
> 1. call the local hospital for their procedure--great suggestion of all who made it!
> 2. check with your Biosafety committee for their recommendations; check whether there's anything in your current BUA that can guide you -- maybe you need to amend your BUA??
> 3. once cleared, give the user his own blades and make sure they only use these
> 4. make sure to clean everything off after each use with unfixed human tissue with 70% EtOH, then 100% EtOh (the latter for complete drying in the chamber)
> 5. if you have a UV lamp in your cryostat, use it after each use for 30 minutes or so.
       

--
Martin Wessendorf, Ph.D.                   office: [phone]
Assoc Prof, Dept Neuroscience                 lab: [phone]
University of Minnesota             Preferred FAX: [phone]
6-145 Jackson Hall, 321 Church St. SE    Dept Fax: [phone]
Minneapolis, MN  55455                    e-mail: [hidden email]

[Mike Tighe]

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I can see situations where someone would want to do Formalin fixed frozen sections. One might be to avoid the harsh chemicals and heat of processing into Paraffin. People often formalin fix and then put tissues through a sucrose gradient and into OCT.

Aerosols could be a concern especially if some one is using those "freeze it" cans of aerosol to cool down embedded tissues. I think you just need an approved protocol for cleaning and cutting the tissue and proper PPE.

>>> "Jurkevic, Aleksandr" <[hidden email]> 8/30/2011 6:54 PM >>>
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Hello All,

This is not quite a microscopy question. At our core we have a sample preparation lab where clients can section their tissue samples. An investigator wants to cut cryosections of human tissue (skin). We are primarily a research facility and not a clinical lab but I do not see a problem letting him to cut the sections providing that samples are properly formalin-fixed and he has approved protocols for his procedures. Am I oversimplifying the issue and are there any biosafety aspects that have to be considered? I would appreciate if you could share your policies on dealing with human tissue samples.

Thank you.


Aleksandr Jurkevic, PhD
Associate Director
Molecular Cytology Core
University of Missouri-Columbia
120 Life Sciences Center
1201 E. Rollins St.
Columbia, MO 65211-7310

Phone:    [phone]
Fax:           [phone]
website  http://www.biotech.missouri.edu/mcc/ 

 

[Ray Gilbert]

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WHO/CDS/CSR/APH/2000.3 - WHO Infection Control Guidelines for Transmissible
Spongiform Encephalopathies (i.e. Prions which do survive formalin fixation) contains the following statement in its table of infectivity

Actual infectivity titres in the various human tissues other than the brain are extremely limited

It doesn't rate skin but finds no detectable activity in muscle, blood or sweat, so prions are highly unlikely to be an issue for your user, however your internal policies and local/national standards and codes of practice are your ultimate arbiter.  

Thanks

Ray


-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Martin Wessendorf
Sent: Thursday, 1 September 2011 4:30 a.m.
To: [hidden email]
Subject: Re: human tissue sections

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Are aerosols from cryostat-sectioned tissue an issue?  I know that when
I cryosection rat brains, the particulates are sufficient to awaken my
rat allergy and set me sneezing.  I would guess that with particulates
containing viruses or prions could also be inhaled.

Martin Wessendorf

On 8/31/2011 11:21 AM, Danielle Crippen wrote:

> The need to cryosection most surely means the tissue won't be fixed...maybe they will need ORO or Betagal staining or something along those lines that requires fresh frozen tissue.
>
> As long as the tissue isn't positive for HIV (or some other virus)which might require additional guidelines, I would approach this situation in the following manner:
>
> 1. call the local hospital for their procedure--great suggestion of all who made it!
> 2. check with your Biosafety committee for their recommendations; check whether there's anything in your current BUA that can guide you -- maybe you need to amend your BUA??
> 3. once cleared, give the user his own blades and make sure they only use these
> 4. make sure to clean everything off after each use with unfixed human tissue with 70% EtOH, then 100% EtOh (the latter for complete drying in the chamber)
> 5. if you have a UV lamp in your cryostat, use it after each use for 30 minutes or so.
       

--
Martin Wessendorf, Ph.D.                   office: [phone]
Assoc Prof, Dept Neuroscience                 lab: [phone]
University of Minnesota             Preferred FAX: [phone]
6-145 Jackson Hall, 321 Church St. SE    Dept Fax: [phone]
Minneapolis, MN  55455                    e-mail: [hidden email]