[3dem] is it possible to place vitrbo in biosafety Class II Bcabinet?

[Wanzhong/He]

Dear all,

Thank Dewight R. Williams, Paul Chipman  Jun Liu, Bill Tivol, Wei Zhang for
sharing their experience in the following !

Here I summarized the solutions for place vitrobot in biosafety cabinet
Class 2 type B:

 It is possible to place the vitrobot into the BSC 2B. However, we need to
solving following problems:
1. the ice contamination due to the air flow (need special care for grid
transfer) 
2. the damage of the electrial coatings of vitrobot (need reflective
coating) 
3. Avoid organics or acids in hood
4. the BSC 2B must be taller than 34² for vitrobot

Others tends to place the vitrobot on bench near the BSC 2A or 2B. Handle
pathogenic sample in BSC 2, but with some special protocol or accessory
attached to the vitrobot for handling  and cleaning. Or just purchased a
cheap home-made plunge freezer and place into the BSC 2B instead.

Best,

Wanzhong



On 1/12/09 11:46 PM, "Williams, Dewight R"
<dewight.r.williams at Vanderbilt.Edu> wrote:

> Dear Wanzong,
>  
> In the Stewart lab at Vanderbilt, our vitrobot resides in a BSC IIB (it looks
> like a Labgaurd 430/435) without any troubles (plenty of room). We are asked
> by FEI to limit the use of our UV sterilization as this will deteriorate the
> vitrobot tubing and electrical coatings. With some effort, reflective coating
> could overcome this limitation, if your biosaftey officer requires UV
> sterilization versus bleach or ethanol. We don't use many organics or acids in
> our hood. Since these can seriously compromise your mechanics on the vitrobot,
> I would contact FEI about compounds they don't recommend. Also, the air flow
> tends to cause ice contamination during transferring from the ethane cup to
> the liquid nitrogen for the newbie, but this can be overcome with practice.
> Hope this helps,
>  
>  
> Dewight  
>  
> Dewight R. Williams Ph.D.
> Department Molecular Physiology and Biophysics
> Vanderbilt University
> 708 Light Hall, 2215 Garland Ave
> Nashville, TN. 37232
> Lab: (615) 322-7898
 
Wanzhong,

Another thing to keep in mind is the interior height of the BSC vs the
height of the Vitrobot. Most BSCs have an interior height of 27" while
the height of our Vitrobot is ~34". Fume hoods usually have a greater
interior height. I think there are a couple BSCs that can be purchased
with an interior height just over 34" that might be sufficient. Anyone
have this set up?


-paul

-- 
Paul Chipman
Director, Biological Electron Microscopy Facility
Purdue University
765-494-1487




Hi Wanzong,
Our Vitrobot is on a lab bench with a small manifold for removing ethane
that can be placed above the location where the gas is used to fill the
cryogen cup.  Once the cryogen has cooled to nearly LN2 temperature, there
is very little gas evolved from it, so there is no need to put the Vitrobot
in a hood.  I would be more worried about contamination of the Vitrobot by
whatever must be used in the biosafety cabinet than by any potential danger
from the flamability of the ethane.
 
Yours,

Bill Tivol, PhD

EM Scientist

Ultrafast EM Facility

Noyes Laboratory, MC 127-72

California Institute of Technology

Pasadena CA 91125

(626) 395-8833

tivol at caltech.edu


Hi, Wanzhong,
 
We have a homemade plunge freezer (you can buy it with ~$5000) to prepare
cryo EM samples inside biosafty cabinet. It works well with limited
training.
 
Jun

> 
> Hi Wanzhong,

Yes, there is a safety issue if vitrobot is put in a biosafety cabinet
because air inside the cabinet is filtered and sent back into the
room. The lafety officer in our university specially pointed out that we
should not use ethane in a biosafety hood. I understand that only little
amount of ethane is used for freezing, so if do it carefully, nothing bad
will happen. But it is a good idea to follow the regulation policy of the
institution.

Best regards,

Wei
========
> Wei Zhang
> 
> From: Wanzhong He
> Sent: Mon 1/12/2009 1:28 AM
> To: 3dem at ncmir.ucsd.edu
> Subject: [3dem] is it possible to place vitrbo in biosafety Class II B
> cabinet?
> 
> Dear all,
>  
>  Due to the limitted room size, it is difficult to fit both biosafety cabinet
> class II A and fume hood ( proposed for placing  vitrobot) into our samll
> sample preparation room. So we are checking whether is possible to combine the
> the biosafety cabinet and fume hood as one?  For example, using Biosafety
> cabinet class II B for placing the vitrobot. Will it be a safety issue using a
> little flammable ethane into the Biosafety cabinet class II B?
>  
> Thanks in advance!
>  
> Wanzhong
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---------------------------
Assistant Professor He Wanzhong :: Cellular Electron Tomography & Cryo-EM
lab :: Blk S3 #02-06 :: National University of Singapore :: Department of
Biological Sciences,  14 Science Drive 4, Singapore 117543 :: 65-6516 7378
(Office) ::65-6516 7068(Lab):: 65-6779 2486 (Fax) ::dbshw at nus.edu.sg (E) ::
www.dbs.nus.edu.sg/staff/hewz.htm  (W) 

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