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BSL-2 specimens with a dipping lens

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Greg Martin-8

BSL-2 specimens with a dipping lens

Hey Folks --

Have any of you had experience dealing with BSL-2 specimens imaged in an open dish with a dipping lens? Adherent BSL-2 cultured cells are less of an issue as they are generally in a covered dish, which is then within an incubator (stage or 'scope enclosure) while being imaged from below on an inverted. But I have users wanting to do BSL-2 work on live, isolated tissue with 2-photon using a high-NA dipping lens, so of course the dish must be open. I take that back -- the Olympus 25X NA 1.05 has a correction collar for a coverglass between lens and specimen so we could make a chamber with a coverglass lid, but that seems pretty complicated and messy (no air between specimen and chamber lid). Anyway...

Are decontamination solutions safe for the lens (I have a question out to Olympus on this as well)? Media spills are the biggest issue and are inevitable. Seems like BSL-2 media spilled onto a 'scope table tapped with hundreds of holes would be a nightmare to clean up properly. Have folks tried putting a tray under the specimen dish (no DIC then) or under the entire 'scope? Any ideas/experiences for dealing with these sorts of experiments would be greatly appreciated!

Be peace! Greg.

Greg Martin

Keck Microscopy Facility
University of Washington School of Medicine
www.depts.washington.edu/keck

And

Lynn and Mike Garvey Cell Imaging Laboratory
University of Washington Institute for Stem Cell and Regenerative Medicine
www.depts.washington.edu/garveyic

206-685-8784 (office)
425-344-2632 (cell)

Phillips, Thomas E.

Re: BSL-2 specimens with a dipping lens

I hope you realize that this isn’t just a decision between you and your client. Using BSL-2 agents would undoubtedly require approval by some committee at your University. They approve specific protocols and locations. Just because my lab is BSL-2 approved, it doesn’t mean I can take my agents next door to the core facility I manage. My lab had to be inspected before I got approval. Presumably your question is fact gathering for the protocol you intend to submit.

Thomas E. Phillips, Ph.D

Professor of Biological Sciences

Director, Molecular Cytology Core

2 Tucker Hall

University of Missouri

Columbia, MO 65211-7400

573-882-4712 (office)

573-882-0123 (fax)

[hidden email]

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Greg Martin
Sent: Thursday, October 29, 2009 5:12 PM
To: [hidden email]
Subject: BSL-2 specimens with a dipping lens

Hey Folks --

Be peace! Greg.

Greg Martin

Keck Microscopy Facility
University of Washington School of Medicine
www.depts.washington.edu/keck

And

Lynn and Mike Garvey Cell Imaging Laboratory
University of Washington Institute for Stem Cell and Regenerative Medicine
www.depts.washington.edu/garveyic

206-685-8784 (office)
425-344-2632 (cell)

Kurt Thorn

Re: BSL-2 specimens with a dipping lens

In reply to this post by Greg Martin-8

I think this is pretty common, actually. I've read a number of
proposals where they propose to decontaminate objectives with 70%
ethanol. Given that we routinely clean objectives with methanol I don't
think this would be a problem. Spills are a potential issue, however,
and I'm not sure that there is a good solution to this.

Kurt

Greg Martin wrote:

> Hey Folks --
>
> Have any of you had experience dealing with BSL-2 specimens imaged
> in an open dish with a dipping lens? Adherent BSL-2 cultured cells
> are less of an issue as they are generally in a covered dish, which is
> then within an incubator (stage or 'scope enclosure) while being
> imaged from below on an inverted. But I have users wanting to do
> BSL-2 work on live, isolated tissue with 2-photon using a high-NA
> dipping lens, so of course the dish must be open. I take that back --
> the Olympus 25X NA 1.05 has a correction collar for a coverglass
> between lens and specimen so we could make a chamber with a coverglass
> lid, but that seems pretty complicated and messy (no air between
> specimen and chamber lid). Anyway...
>
> Are decontamination solutions safe for the lens (I have a question
> out to Olympus on this as well)? Media spills are the biggest issue
> and are inevitable. Seems like BSL-2 media spilled onto a 'scope
> table tapped with hundreds of holes would be a nightmare to clean up
> properly. Have folks tried putting a tray under the specimen dish (no
> DIC then) or under the entire 'scope? Any ideas/experiences
> for dealing with these sorts of experiments would be greatly appreciated!
>
> Be peace! Greg.
>
> Greg Martin
>
> Keck Microscopy Facility
> University of Washington School of Medicine
> www.depts.washington.edu/keck <http://www.depts.washington.edu/keck>
>
> And
>
> Lynn and Mike Garvey Cell Imaging Laboratory
> University of Washington Institute for Stem Cell and Regenerative Medicine
> www.depts.washington.edu/garveyic
> <http://www.depts.washington.edu/garveyic>
>
> 206-685-8784 (office)
> 425-344-2632 (cell)

Caroline Bass

Re: BSL-2 specimens with a dipping lens

In reply to this post by Greg Martin-8

Re: BSL-2 specimens with a dipping lens Hello Greg,

I can’t give you any specific advice since you haven’t stated the nature of the biohazard, but I have a lot of experiments working with biosafety issues. I think the most important aspect is that you need a biosafety protocol in place, or the person who is the PI on the project needs to add your facility to their protocol. BSL-2 generally requires a biosafety cabinet, but there are ways to get around this. I do a lot of stereotaxic surgeries, and they would prefer that they be done in a biosafety cabinet, but this is virtually impossible (unless I stick my head in there which kind of defeats the purpose!) so I have an exception for this aspect of my work.

Basically, you should be able to do this, but a biosafety application has to be made (or an existing one amended). You have to list the nature of the biohazard and how you will handle spills and possible contamination, emergency numbers, etc. If the committee doesn’t like it, they will tell you exactly what you need to change. Every institute is different, what may be considered BSL-1 at my institute could be BSL-2 at yours, so you really need to talk with your biosafety officer about your options. Think of it as an ACUC protocol, but for biohazards instead of animals!

Caroline Bass

On 10/29/09 6:11 PM, "Greg Martin" <keck@...> wrote:

Hey Folks --

Have any of you had experience dealing with BSL-2 specimens imaged in an open dish with a dipping lens? Adherent BSL-2 cultured cells are less of an issue as they are generally in a covered dish, which is then within an incubator (stage or 'scope enclosure) while being imaged from below on an inverted. But I have users wanting to do BSL-2 work on live, isolated tissue with 2-photon using a high-NA dipping lens, so of course the dish must be open. I take that back -- the Olympus 25X NA 1.05 has a correction collar for a coverglass between lens and specimen so we could make a chamber with a coverglass lid, but that seems pretty complicated and messy (no air between specimen and chamber lid). Anyway...

Are decontamination solutions safe for the lens (I have a question out to Olympus on this as well)? Media spills are the biggest issue and are inevitable. Seems like BSL-2 media spilled onto a 'scope table tapped with hundreds of holes would be a nightmare to clean up properly. Have folks tried putting a tray under the specimen dish (no DIC then) or under the entire 'scope? Any ideas/experiences for dealing with these sorts of experiments would be greatly appreciated!

Be peace! Greg.

Greg Martin

Keck Microscopy Facility
University of Washington School of Medicine
www.depts.washington.edu/keck <http://www.depts.washington.edu/keck>

And

Lynn and Mike Garvey Cell Imaging Laboratory
University of Washington Institute for Stem Cell and Regenerative Medicine
www.depts.washington.edu/garveyic <http://www.depts.washington.edu/garveyic>

206-685-8784 (office)
425-344-2632 (cell)

Greg Martin-8

Re: BSL-2 specimens with a dipping lens

In reply to this post by Phillips, Thomas E.

Hey Tom --

Yep, I'm well aware. We need to allow BSL-2 in this particular shared facility, and yes, the biosafety people of my university have inspected the microscopes and approved putting BSL-2 specimens on them. Meaning: gloves, gowns, signage and spill kit. Doesn't give me a lot of confidence. No protocols were requested. As you point out, users need approval of their own protocols for handling these specimens in their own lab long before they get to the microscope --but I don't know if their protocols specifically include the use of the microscopes in this facility. I would tend to think they often do not, and would never assume that they spelled things out about the microscopy. So, if their personal protocols have been approved, and my shared microscopes have been approved, are we good to go?

How has this been dealt with at other institutions? There are procedures in place at my institution for amending a previously approved protocol, maybe that's the way to go to include the microscopes if they weren't part of the original proposal.....

Be peace! Greg.

Greg Martin

Keck Microscopy Facility
University of Washington School of Medicine
www.depts.washington.edu/keck

And

Lynn and Mike Garvey Cell Imaging Laboratory
University of Washington Institute for Stem Cell and Regenerative Medicine
www.depts.washington.edu/garveyic

206-685-8784 (office)
425-344-2632 (cell)

----- Original Message -----

From: [hidden email]

To: [hidden email]

Sent: Thursday, October 29, 2009 3:17 PM

Subject: Re: BSL-2 specimens with a dipping lens

I hope you realize that this isnt just a decision between you and your client. Using BSL-2 agents would undoubtedly require approval by some committee at your University. They approve specific protocols and locations. Just because my lab is BSL-2 approved, it doesnt mean I can take my agents next door to the core facility I manage. My lab had to be inspected before I got approval. Presumably your question is fact gathering for the protocol you intend to submit.

Thomas E. Phillips, Ph.D

Professor of Biological Sciences

Director, Molecular Cytology Core

2 Tucker Hall

University of Missouri

Columbia, MO 65211-7400

573-882-4712 (office)

573-882-0123 (fax)

[hidden email]

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Greg Martin
Sent: Thursday, October 29, 2009 5:12 PM
To: [hidden email]
Subject: BSL-2 specimens with a dipping lens

Hey Folks --

Have any of you had experience dealing with BSL-2 specimens imaged in an open dish with a dipping lens? Adherent BSL-2 cultured cells are less of an issue as they are generally in a covered dish, which is then within an incubator (stage or 'scope enclosure) while being imaged from below on an inverted. But I have users wanting to do BSL-2 work on live, isolated tissue with 2-photon using a high-NA dipping lens, so of course the dish must be open. I take that back -- the Olympus 25X NA 1.05 has a correction collar for a coverglass between lens and specimen so we could make a chamber with a coverglass lid, but that seems pretty complicated and messy (no air between specimen and chamber lid). Anyway...

Are decontamination solutions safe for the lens (I have a question out to Olympus on this as well)? Media spills are the biggest issue and are inevitable. Seems like BSL-2 media spilled onto a 'scope table tapped with hundreds of holes would be a nightmare to clean up properly. Have folks tried putting a tray under the specimen dish (no DIC then) or under the entire 'scope? Any ideas/experiences for dealing with these sorts of experiments would be greatly appreciated!

Be peace! Greg.

Greg Martin

Keck Microscopy Facility
University of Washington School of Medicine
www.depts.washington.edu/keck

And

Lynn and Mike Garvey Cell Imaging Laboratory
University of Washington Institute for Stem Cell and Regenerative Medicine
www.depts.washington.edu/garveyic

206-685-8784 (office)
425-344-2632 (cell)