BSL3 microscopy

[Watkins, Simon C]

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Folks, I am about to install an advanced imaging system (sweptfield confocal and widefield imaging combo) in a biosafety level 3 facility.  To work in the facility one has to suit up and wear a rigid face mask, as the cells/pathogens will be on the scope and may be open to the environment.  This means that the eyepieces of the scope are essentially useless.  I wonder if any other listers have dealt with this problem and what their solution was? Obviously the widefield camera will help a lot, but it doesnt allow survey of the full field of view, as we are doing mostly flourescence a video camera isnt much use.... back in the day, there were some screen solutions....
Looking for creative ideas
S.

Simon C. Watkins Ph.D, FRC Path
Professor and Vice Chair Cell Biology and Physiology
Professor Immunology Director Center for Biologic Imaging
BSTS 225
University of Pittsburgh
3500 Terrace St
Pittsburgh PA 15261
412-352-2277
www.cbi.pitt.edu<http://www.cbi.pitt.edu>

[Peter Humphreys]

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Hi, vision engineering produce isis eyepieces which have a large exit
pupil, we've used them through the glass window of a cabinet.
they work using a spinning projection screen, so their use in
fluorescence may be in question as it may not be bright enough. I'd be
very interested if they work for fluorescence.

Peter

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Folks, I am about to install an advanced imaging system (sweptfield confocal and widefield imaging combo) in a biosafety level 3 facility.  To work in the facility one has to suit up and wear a rigid face mask, as the cells/pathogens will be on the scope and may be open to the environment.  This means that the eyepieces of the scope are essentially useless.  I wonder if any other listers have dealt with this problem and what their solution was? Obviously the widefield camera will help a lot, but it doesnt allow survey of the full field of view, as we are doing mostly flourescence a video camera isnt much use.... back in the day, there were some screen solutions....
> Looking for creative ideas
> S.
>
> Simon C. Watkins Ph.D, FRC Path
> Professor and Vice Chair Cell Biology and Physiology
> Professor Immunology Director Center for Biologic Imaging
> BSTS 225
> University of Pittsburgh
> 3500 Terrace St
> Pittsburgh PA 15261
> 412-352-2277
> www.cbi.pitt.edu<http://www.cbi.pitt.edu>

[Johannes Helm]

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Good afternoon,

the straight forward way, if an additional camera port is available on the
microscope, might be to mount a CCD camera and between the microscope and
the CCD camera a C-mount adapter including a 0.32x or 0.5x scaler. The
disadvantage is that the image produced by the camera will show "black
corners" around the image of the FOV since the entire FOV is projected
onto the CCD chip, which is rectangular, while the FOV is cicular in
shape.

If you canNOT proceed this way:
You might try what just is an idea and I do not know whether this would work:

The eyepiece, as many of the group members know, is used as a
magnification glass to observe the real intermediate image, which, in
most, possibly all, standards is located 10mm inside the tube which holds
the eyepiece(s), distance measured from the edge to which the eyepiece is
set.

If your preps produce images with sufficiently large contrasts (otherwise
use DIC or PC or another contrast method):
If you would place (a) circular screen(s) made from a whitish,
structureless material or possibly frozen glass into the tube(s) holding
the eyepiece(s) so that the screens are positioned 10mm inside the
tube(s), you would be able to view the real intermediate on these small
screens. Of course, the image is much too small for eye observation, but
you might place a digital photoapparatus in "near view mode" so that it
will image this real intermediate image. I've just tried with my Olympus
my 810, and it easily images the 25mm which is, as far as I know, the
largest diameter of the FOV in the plane of the real intermediate image.


Best wishes,

Johannes

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Folks, I am about to install an advanced imaging system (sweptfield
> confocal and widefield imaging combo) in a biosafety level 3 facility.  To
> work in the facility one has to suit up and wear a rigid face mask, as the
> cells/pathogens will be on the scope and may be open to the environment.
> This means that the eyepieces of the scope are essentially useless.  I
> wonder if any other listers have dealt with this problem and what their
> solution was? Obviously the widefield camera will help a lot, but it
> doesnt allow survey of the full field of view, as we are doing mostly
> flourescence a video camera isnt much use.... back in the day, there were
> some screen solutions....
> Looking for creative ideas
> S.
>
> Simon C. Watkins Ph.D, FRC Path
> Professor and Vice Chair Cell Biology and Physiology
> Professor Immunology Director Center for Biologic Imaging
> BSTS 225
> University of Pittsburgh
> 3500 Terrace St
> Pittsburgh PA 15261
> 412-352-2277
> www.cbi.pitt.edu<http://www.cbi.pitt.edu>
>

--
P. Johannes Helm

Voice: ([phone] (office)
Fax: [phone] (office)

[Emmanuel Gustin]

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I've never used them myself, but a number of manufacturers sell eyepiece

cameras, which can be inserted in the place of a standard eyepiece. You
may
need to buy one which supports a fairly high frame rate, and indeed
sensitivity on these simple cameras may be an issue.

On our Leica confocal, which also is in BSL-3, you can easily remove the

outer rings from the eyepieces. This exposes the surface of the lenses,
which are surrounded by soft rings, so that you can indeed use them
while
wearing standard safety glasses: We don't use full face masks, however.


Best Regards,

Emmanuel



-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]]
On Behalf Of Johannes Helm
Sent: vrijdag 14 januari 2011 16:49
To: [hidden email]
Subject: Re: BSL3 microscopy

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Good afternoon,

the straight forward way, if an additional camera port is available on
the
microscope, might be to mount a CCD camera and between the microscope
and
the CCD camera a C-mount adapter including a 0.32x or 0.5x scaler. The
disadvantage is that the image produced by the camera will show "black
corners" around the image of the FOV since the entire FOV is projected
onto the CCD chip, which is rectangular, while the FOV is cicular in
shape.

If you canNOT proceed this way:
You might try what just is an idea and I do not know whether this would
work:

The eyepiece, as many of the group members know, is used as a
magnification glass to observe the real intermediate image, which, in
most, possibly all, standards is located 10mm inside the tube which
holds
the eyepiece(s), distance measured from the edge to which the eyepiece
is
set.

If your preps produce images with sufficiently large contrasts
(otherwise
use DIC or PC or another contrast method):
If you would place (a) circular screen(s) made from a whitish,
structureless material or possibly frozen glass into the tube(s) holding
the eyepiece(s) so that the screens are positioned 10mm inside the
tube(s), you would be able to view the real intermediate on these small
screens. Of course, the image is much too small for eye observation, but
you might place a digital photoapparatus in "near view mode" so that it
will image this real intermediate image. I've just tried with my Olympus
my 810, and it easily images the 25mm which is, as far as I know, the
largest diameter of the FOV in the plane of the real intermediate image.


Best wishes,

Johannes


> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Folks, I am about to install an advanced imaging system (sweptfield
> confocal and widefield imaging combo) in a biosafety level 3 facility.
To
> work in the facility one has to suit up and wear a rigid face mask, as
the
> cells/pathogens will be on the scope and may be open to the
environment.
> This means that the eyepieces of the scope are essentially useless.  I
> wonder if any other listers have dealt with this problem and what
their
> solution was? Obviously the widefield camera will help a lot, but it
> doesnt allow survey of the full field of view, as we are doing mostly
> flourescence a video camera isnt much use.... back in the day, there
were

> some screen solutions....
> Looking for creative ideas
> S.
>
> Simon C. Watkins Ph.D, FRC Path
> Professor and Vice Chair Cell Biology and Physiology
> Professor Immunology Director Center for Biologic Imaging
> BSTS 225
> University of Pittsburgh
> 3500 Terrace St
> Pittsburgh PA 15261
> 412-352-2277
> www.cbi.pitt.edu<http://www.cbi.pitt.edu>
>

--
P. Johannes Helm

Voice: ([phone] (office)
Fax: [phone] (office)

[Caroline Bass]

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Hi,

I've worked in a BSL3 lab before, and did use a general inverted
microscope. Are you sure that the specimen can't be contained and that you
need the face shield? The purpose of a BSL3 facility is to maintain
negative air pressure in case of an incident. A face shield protects
against accidental splashes but doesn't do anything for an aerosolized or
airborne agent (they easily get around a mask). So it seems if there is a
way to contain the specimen either in a sealed environment (flask, or
slide chamber) or to set up a shield between the user and the specimen,
that would serve the same purpose.

Also, what about a monitor situation? If you can contain the specimen from
the user then it seems your typical confocal setup, where most of the work
is done on a computer, contained within the BSL3 facility should be
sufficient.

Caroline Bass




On 1/14/11 9:32 AM, "Watkins, Simon C" <[hidden email]> wrote:

>*****
>To join, leave or search the confocal microscopy listserv, go to:
>http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>*****
>
>Folks, I am about to install an advanced imaging system (sweptfield
>confocal and widefield imaging combo) in a biosafety level 3 facility.
>To work in the facility one has to suit up and wear a rigid face mask, as
>the cells/pathogens will be on the scope and may be open to the
>environment.  This means that the eyepieces of the scope are essentially
>useless.  I wonder if any other listers have dealt with this problem and
>what their solution was? Obviously the widefield camera will help a lot,
>but it doesnt allow survey of the full field of view, as we are doing
>mostly flourescence a video camera isnt much use.... back in the day,
>there were some screen solutions....
>Looking for creative ideas
>S.
>
>Simon C. Watkins Ph.D, FRC Path
>Professor and Vice Chair Cell Biology and Physiology
>Professor Immunology Director Center for Biologic Imaging
>BSTS 225
>University of Pittsburgh
>3500 Terrace St
>Pittsburgh PA 15261
>412-352-2277
>www.cbi.pitt.edu<http://www.cbi.pitt.edu>

[Steffen Dietzel]

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Simon,

if you put a little screen in front of the eye piece, some centimeters
away, at the right distance you'll get a sharp image. You can
demonstrate the effect easily with a bright field microscope and a piece
of paper. You probably could borrow a screen from somebody teaching
basic optics at the physics department.

With similar strategy, you could mount a digital consumer camera in
front of the eye piece. A respective adapter is shown in the image on
this page (bottom image):
http://www.doerrfoto.de/Produkte/Article.aspx?productline=192&article=566556
(make sure its all in one line in your browser's address field)
The adapter is clamped around the eye piece and the camera then
centered. I got such an adapter from a local photography/optics shop.
Works nice for bright field, e.g. for preparation microscopes. No
experience with fluorescence though.

Of course, for fluorescence sensitivity will be an issue in both cases
and I wouldn't be too optimistic. But since the attached costs are
rather modest, maybe it's worth trying.

Steffen



On 14.01.2011 15:32, Watkins, Simon C wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Folks, I am about to install an advanced imaging system (sweptfield confocal and widefield imaging combo) in a biosafety level 3 facility.  To work in the facility one has to suit up and wear a rigid face mask, as the cells/pathogens will be on the scope and may be open to the environment.  This means that the eyepieces of the scope are essentially useless.  I wonder if any other listers have dealt with this problem and what their solution was? Obviously the widefield camera will help a lot, but it doesnt allow survey of the full field of view, as we are doing mostly flourescence a video camera isnt much use.... back in the day, there were some screen solutions....
> Looking for creative ideas
> S.
>
> Simon C. Watkins Ph.D, FRC Path
> Professor and Vice Chair Cell Biology and Physiology
> Professor Immunology Director Center for Biologic Imaging
> BSTS 225
> University of Pittsburgh
> 3500 Terrace St
> Pittsburgh PA 15261
> 412-352-2277
> www.cbi.pitt.edu<http://www.cbi.pitt.edu>
>

--
------------------------------------------------------------
Steffen Dietzel, PD Dr. rer. nat
Ludwig-Maximilians-Universität München
Walter-Brendel-Zentrum für experimentelle Medizin (WBex)
Head of light microscopy

Mail room:
Marchioninistr. 15, D-81377 München

Building location:
Marchioninistr. 27,  München-Großhadern

[Craig Brideau]

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I've had good success with a cheap machine vision camera and a standard tube
lens.  If you have a camera port the tube lens will probably already be
installed there.  The trick will be getting a wide enough field of view.
 Generally a larger CCD array on the camera will help with this.

Craig


On Fri, Jan 14, 2011 at 10:12 AM, Steffen Dietzel <[hidden email]>wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Simon,
>
> if you put a little screen in front of the eye piece, some centimeters
> away, at the right distance you'll get a sharp image. You can demonstrate
> the effect easily with a bright field microscope and a piece of paper. You
> probably could borrow a screen from somebody teaching basic optics at the
> physics department.
>
> With similar strategy, you could mount a digital consumer camera in front
> of the eye piece. A respective adapter is shown in the image on this page
> (bottom image):
>
> http://www.doerrfoto.de/Produkte/Article.aspx?productline=192&article=566556
> (make sure its all in one line in your browser's address field)
> The adapter is clamped around the eye piece and the camera then centered. I
> got such an adapter from a local photography/optics shop. Works nice for
> bright field, e.g. for preparation microscopes. No experience with
> fluorescence though.
>
> Of course, for fluorescence sensitivity will be an issue in both cases and
> I wouldn't be too optimistic. But since the attached costs are rather
> modest, maybe it's worth trying.
>
> Steffen
>
>
>
>
> On 14.01.2011 15:32, Watkins, Simon C wrote:
>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> *****
>>
>> Folks, I am about to install an advanced imaging system (sweptfield
>> confocal and widefield imaging combo) in a biosafety level 3 facility.  To
>> work in the facility one has to suit up and wear a rigid face mask, as the
>> cells/pathogens will be on the scope and may be open to the environment.
>>  This means that the eyepieces of the scope are essentially useless.  I
>> wonder if any other listers have dealt with this problem and what their
>> solution was? Obviously the widefield camera will help a lot, but it doesnt
>> allow survey of the full field of view, as we are doing mostly flourescence
>> a video camera isnt much use.... back in the day, there were some screen
>> solutions....
>> Looking for creative ideas
>> S.
>>
>> Simon C. Watkins Ph.D, FRC Path
>> Professor and Vice Chair Cell Biology and Physiology
>> Professor Immunology Director Center for Biologic Imaging
>> BSTS 225
>> University of Pittsburgh
>> 3500 Terrace St
>> Pittsburgh PA 15261
>> 412-352-2277
>> www.cbi.pitt.edu<http://www.cbi.pitt.edu>
>>
>>
>
> --
> ------------------------------------------------------------
> Steffen Dietzel, PD Dr. rer. nat
> Ludwig-Maximilians-Universität München
> Walter-Brendel-Zentrum für experimentelle Medizin (WBex)
> Head of light microscopy
>
> Mail room:
> Marchioninistr. 15, D-81377 München
>
> Building location:
> Marchioninistr. 27,  München-Großhadern
>

[Watkins, Simon C]

*****
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*****

This little question is proving quite fruitful for discussion, this is not a face shield but totally enclosed nonflexible mask with filtered air.  to stop any accidents aerosolized or otherwise, the bugs are right on the edge of a level 4 in my mind. we tried screens this am, frosted glass, not enough light.  video cameras not enough light, CCD cameras with a reducing adaptor will work but difficult and expensive.  The most interesting suggestion so far has been the vision engineering eyepieces.. They superficially look perfect http://www.visioneng.us/isis-ergonomic-microscope-eyepieces-overview.php but.... they stopped selling them 6 months ago... (still on the website, and they are hunting for some)


Simon C. Watkins Ph.D, FRC Path
Professor and Vice Chair Cell Biology and Physiology
Professor Immunology Director Center for Biologic Imaging
BSTS 225
University of Pittsburgh
3500 Terrace St
Pittsburgh PA 15261
412-352-2277
www.cbi.pitt.edu


-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Caroline Bass
Sent: Friday, January 14, 2011 11:10 AM
To: [hidden email]
Subject: Re: BSL3 microscopy

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Hi,

I've worked in a BSL3 lab before, and did use a general inverted
microscope. Are you sure that the specimen can't be contained and that you
need the face shield? The purpose of a BSL3 facility is to maintain
negative air pressure in case of an incident. A face shield protects
against accidental splashes but doesn't do anything for an aerosolized or
airborne agent (they easily get around a mask). So it seems if there is a
way to contain the specimen either in a sealed environment (flask, or
slide chamber) or to set up a shield between the user and the specimen,
that would serve the same purpose.

Also, what about a monitor situation? If you can contain the specimen from
the user then it seems your typical confocal setup, where most of the work
is done on a computer, contained within the BSL3 facility should be
sufficient.

Caroline Bass




On 1/14/11 9:32 AM, "Watkins, Simon C" <[hidden email]> wrote:

>*****
>To join, leave or search the confocal microscopy listserv, go to:
>http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>*****
>
>Folks, I am about to install an advanced imaging system (sweptfield
>confocal and widefield imaging combo) in a biosafety level 3 facility.
>To work in the facility one has to suit up and wear a rigid face mask, as
>the cells/pathogens will be on the scope and may be open to the
>environment.  This means that the eyepieces of the scope are essentially
>useless.  I wonder if any other listers have dealt with this problem and
>what their solution was? Obviously the widefield camera will help a lot,
>but it doesnt allow survey of the full field of view, as we are doing
>mostly flourescence a video camera isnt much use.... back in the day,
>there were some screen solutions....
>Looking for creative ideas
>S.
>
>Simon C. Watkins Ph.D, FRC Path
>Professor and Vice Chair Cell Biology and Physiology
>Professor Immunology Director Center for Biologic Imaging
>BSTS 225
>University of Pittsburgh
>3500 Terrace St
>Pittsburgh PA 15261
>412-352-2277
>www.cbi.pitt.edu<http://www.cbi.pitt.edu>

[leoncio vergara]

*****
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The purpose of the negative pressure in the BSL3 room is not to protect the users inside the room but to prevent escape of aerosols outside of the room. The full face shield is part of a complete suit, not just a face mask as Simon points out, the purpose of the suit is to protect the user in case of accident happen that releases aerosols, you can encapsulate the sample in a box while in the microscope, but the sample has to get there from the biosafety cabinet, there is when accidents can happen...

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Watkins, Simon C
Sent: Friday, January 14, 2011 11:55 AM
To: [hidden email]
Subject: Re: BSL3 microscopy

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

This little question is proving quite fruitful for discussion, this is not a face shield but totally enclosed nonflexible mask with filtered air.  to stop any accidents aerosolized or otherwise, the bugs are right on the edge of a level 4 in my mind. we tried screens this am, frosted glass, not enough light.  video cameras not enough light, CCD cameras with a reducing adaptor will work but difficult and expensive.  The most interesting suggestion so far has been the vision engineering eyepieces.. They superficially look perfect http://www.visioneng.us/isis-ergonomic-microscope-eyepieces-overview.php but.... they stopped selling them 6 months ago... (still on the website, and they are hunting for some)


Simon C. Watkins Ph.D, FRC Path
Professor and Vice Chair Cell Biology and Physiology
Professor Immunology Director Center for Biologic Imaging
BSTS 225
University of Pittsburgh
3500 Terrace St
Pittsburgh PA 15261
412-352-2277
www.cbi.pitt.edu


-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Caroline Bass
Sent: Friday, January 14, 2011 11:10 AM
To: [hidden email]
Subject: Re: BSL3 microscopy

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Hi,

I've worked in a BSL3 lab before, and did use a general inverted
microscope. Are you sure that the specimen can't be contained and that you
need the face shield? The purpose of a BSL3 facility is to maintain
negative air pressure in case of an incident. A face shield protects
against accidental splashes but doesn't do anything for an aerosolized or
airborne agent (they easily get around a mask). So it seems if there is a
way to contain the specimen either in a sealed environment (flask, or
slide chamber) or to set up a shield between the user and the specimen,
that would serve the same purpose.

Also, what about a monitor situation? If you can contain the specimen from
the user then it seems your typical confocal setup, where most of the work
is done on a computer, contained within the BSL3 facility should be
sufficient.

Caroline Bass




On 1/14/11 9:32 AM, "Watkins, Simon C" <[hidden email]> wrote:

>*****
>To join, leave or search the confocal microscopy listserv, go to:
>http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>*****
>
>Folks, I am about to install an advanced imaging system (sweptfield
>confocal and widefield imaging combo) in a biosafety level 3 facility.
>To work in the facility one has to suit up and wear a rigid face mask, as
>the cells/pathogens will be on the scope and may be open to the
>environment.  This means that the eyepieces of the scope are essentially
>useless.  I wonder if any other listers have dealt with this problem and
>what their solution was? Obviously the widefield camera will help a lot,
>but it doesnt allow survey of the full field of view, as we are doing
>mostly flourescence a video camera isnt much use.... back in the day,
>there were some screen solutions....
>Looking for creative ideas
>S.
>
>Simon C. Watkins Ph.D, FRC Path
>Professor and Vice Chair Cell Biology and Physiology
>Professor Immunology Director Center for Biologic Imaging
>BSTS 225
>University of Pittsburgh
>3500 Terrace St
>Pittsburgh PA 15261
>412-352-2277
>www.cbi.pitt.edu<http://www.cbi.pitt.edu>

[Craig Brideau]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Ideally you want the suits to be an overpressure system, and the room to be
negative pressure.  That way if a suit is breeched air blows OUT of it into
the room, hopefully preventing anything from entering the suit.  Also, if
the room is breeched (a leak or something) air is drawn INTO the room, again
hopefully containing anything pathological...
I can see how the masks typically used in the suits can be a bit problem for
trying to use a microscope.

Craig


On Fri, Jan 14, 2011 at 11:00 AM, Vergara, Leoncio A. <[hidden email]>wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> The purpose of the negative pressure in the BSL3 room is not to protect the
> users inside the room but to prevent escape of aerosols outside of the room.
> The full face shield is part of a complete suit, not just a face mask as
> Simon points out, the purpose of the suit is to protect the user in case of
> accident happen that releases aerosols, you can encapsulate the sample in a
> box while in the microscope, but the sample has to get there from the
> biosafety cabinet, there is when accidents can happen...
>
> -----Original Message-----
> From: Confocal Microscopy List [mailto:[hidden email]]
> On Behalf Of Watkins, Simon C
> Sent: Friday, January 14, 2011 11:55 AM
> To: [hidden email]
> Subject: Re: BSL3 microscopy
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> This little question is proving quite fruitful for discussion, this is not
> a face shield but totally enclosed nonflexible mask with filtered air.  to
> stop any accidents aerosolized or otherwise, the bugs are right on the edge
> of a level 4 in my mind. we tried screens this am, frosted glass, not enough
> light.  video cameras not enough light, CCD cameras with a reducing adaptor
> will work but difficult and expensive.  The most interesting suggestion so
> far has been the vision engineering eyepieces.. They superficially look
> perfect
> http://www.visioneng.us/isis-ergonomic-microscope-eyepieces-overview.phpbut.... they stopped selling them 6 months ago... (still on the website, and
> they are hunting for some)
>
>
> Simon C. Watkins Ph.D, FRC Path
> Professor and Vice Chair Cell Biology and Physiology
> Professor Immunology Director Center for Biologic Imaging
> BSTS 225
> University of Pittsburgh
> 3500 Terrace St
> Pittsburgh PA 15261
> 412-352-2277
> www.cbi.pitt.edu
>
>
> -----Original Message-----
> From: Confocal Microscopy List [mailto:[hidden email]]
> On Behalf Of Caroline Bass
> Sent: Friday, January 14, 2011 11:10 AM
> To: [hidden email]
> Subject: Re: BSL3 microscopy
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hi,
>
> I've worked in a BSL3 lab before, and did use a general inverted
> microscope. Are you sure that the specimen can't be contained and that you
> need the face shield? The purpose of a BSL3 facility is to maintain
> negative air pressure in case of an incident. A face shield protects
> against accidental splashes but doesn't do anything for an aerosolized or
> airborne agent (they easily get around a mask). So it seems if there is a
> way to contain the specimen either in a sealed environment (flask, or
> slide chamber) or to set up a shield between the user and the specimen,
> that would serve the same purpose.
>
> Also, what about a monitor situation? If you can contain the specimen from
> the user then it seems your typical confocal setup, where most of the work
> is done on a computer, contained within the BSL3 facility should be
> sufficient.
>
> Caroline Bass
>
>
>
>
> On 1/14/11 9:32 AM, "Watkins, Simon C" <[hidden email]> wrote:
>
> >*****
> >To join, leave or search the confocal microscopy listserv, go to:
> >http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> >*****
> >
> >Folks, I am about to install an advanced imaging system (sweptfield
> >confocal and widefield imaging combo) in a biosafety level 3 facility.
> >To work in the facility one has to suit up and wear a rigid face mask, as
> >the cells/pathogens will be on the scope and may be open to the
> >environment.  This means that the eyepieces of the scope are essentially
> >useless.  I wonder if any other listers have dealt with this problem and
> >what their solution was? Obviously the widefield camera will help a lot,
> >but it doesnt allow survey of the full field of view, as we are doing
> >mostly flourescence a video camera isnt much use.... back in the day,
> >there were some screen solutions....
> >Looking for creative ideas
> >S.
> >
> >Simon C. Watkins Ph.D, FRC Path
> >Professor and Vice Chair Cell Biology and Physiology
> >Professor Immunology Director Center for Biologic Imaging
> >BSTS 225
> >University of Pittsburgh
> >3500 Terrace St
> >Pittsburgh PA 15261
> >412-352-2277
> >www.cbi.pitt.edu<http://www.cbi.pitt.edu>
>

[Caroline Bass]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

The point is a physical separation of user from specimen. Is there a way to separate the main body of the microscope, that will hold the specimen, from the eyepieces? A sort of mini biosafety cabinet? You can always fabricate an transport enclosure for transfer of the specimen and guard against accidents.

What organisms are you working with, and what specimens/procedures are you doing? I've worked in two different BSL-3 labs and both could easily accommodate a microscope. I'm also curious what you are working with/procedures outside the biosafety cabinets that would necessitate a respirator (just curious). Is this a BSL3+ facility (if such a beast exists, I've worked in a BSL2+).

Caroline

 

On Jan 14, 2011, at 1:00 PM, "Vergara, Leoncio A." <[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> The purpose of the negative pressure in the BSL3 room is not to protect the users inside the room but to prevent escape of aerosols outside of the room. The full face shield is part of a complete suit, not just a face mask as Simon points out, the purpose of the suit is to protect the user in case of accident happen that releases aerosols, you can encapsulate the sample in a box while in the microscope, but the sample has to get there from the biosafety cabinet, there is when accidents can happen...
>
> -----Original Message-----
> From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Watkins, Simon C
> Sent: Friday, January 14, 2011 11:55 AM
> To: [hidden email]
> Subject: Re: BSL3 microscopy
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> This little question is proving quite fruitful for discussion, this is not a face shield but totally enclosed nonflexible mask with filtered air.  to stop any accidents aerosolized or otherwise, the bugs are right on the edge of a level 4 in my mind. we tried screens this am, frosted glass, not enough light.  video cameras not enough light, CCD cameras with a reducing adaptor will work but difficult and expensive.  The most interesting suggestion so far has been the vision engineering eyepieces.. They superficially look perfect http://www.visioneng.us/isis-ergonomic-microscope-eyepieces-overview.php but.... they stopped selling them 6 months ago... (still on the website, and they are hunting for some)
>
>
> Simon C. Watkins Ph.D, FRC Path
> Professor and Vice Chair Cell Biology and Physiology
> Professor Immunology Director Center for Biologic Imaging
> BSTS 225
> University of Pittsburgh
> 3500 Terrace St
> Pittsburgh PA 15261
> 412-352-2277
> www.cbi.pitt.edu
>
>
> -----Original Message-----
> From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Caroline Bass
> Sent: Friday, January 14, 2011 11:10 AM
> To: [hidden email]
> Subject: Re: BSL3 microscopy
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hi,
>
> I've worked in a BSL3 lab before, and did use a general inverted
> microscope. Are you sure that the specimen can't be contained and that you
> need the face shield? The purpose of a BSL3 facility is to maintain
> negative air pressure in case of an incident. A face shield protects
> against accidental splashes but doesn't do anything for an aerosolized or
> airborne agent (they easily get around a mask). So it seems if there is a
> way to contain the specimen either in a sealed environment (flask, or
> slide chamber) or to set up a shield between the user and the specimen,
> that would serve the same purpose.
>
> Also, what about a monitor situation? If you can contain the specimen from
> the user then it seems your typical confocal setup, where most of the work
> is done on a computer, contained within the BSL3 facility should be
> sufficient.
>
> Caroline Bass
>
>
>
>
> On 1/14/11 9:32 AM, "Watkins, Simon C" <[hidden email]> wrote:
>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> *****
>>
>> Folks, I am about to install an advanced imaging system (sweptfield
>> confocal and widefield imaging combo) in a biosafety level 3 facility.
>> To work in the facility one has to suit up and wear a rigid face mask, as
>> the cells/pathogens will be on the scope and may be open to the
>> environment.  This means that the eyepieces of the scope are essentially
>> useless.  I wonder if any other listers have dealt with this problem and
>> what their solution was? Obviously the widefield camera will help a lot,
>> but it doesnt allow survey of the full field of view, as we are doing
>> mostly flourescence a video camera isnt much use.... back in the day,
>> there were some screen solutions....
>> Looking for creative ideas
>> S.
>>
>> Simon C. Watkins Ph.D, FRC Path
>> Professor and Vice Chair Cell Biology and Physiology
>> Professor Immunology Director Center for Biologic Imaging
>> BSTS 225
>> University of Pittsburgh
>> 3500 Terrace St
>> Pittsburgh PA 15261
>> 412-352-2277
>> www.cbi.pitt.edu<http://www.cbi.pitt.edu>

[Peter Humphreys]

We just demo'd some ivis pieces through Olympus uk if that helps. Peter
Sent from my HTC

----- Reply message -----
From: "Watkins, Simon C" <[hidden email]>
To: <[hidden email]>
Subject: BSL3 microscopy
Date: Fri, Jan 14, 2011 17:54


*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

This little question is proving quite fruitful for discussion, this is not a face shield but totally enclosed nonflexible mask with filtered air.  to stop any accidents aerosolized or otherwise, the bugs are right on the edge of a level 4 in my mind. we tried screens this am, frosted glass, not enough light.  video cameras not enough light, CCD cameras with a reducing adaptor will work but difficult and expensive.  The most interesting suggestion so far has been the vision engineering eyepieces.. They superficially look perfect http://www.visioneng.us/isis-ergonomic-microscope-eyepieces-overview.php but.... they stopped selling them 6 months ago... (still on the website, and they are hunting for some)


Simon C. Watkins Ph.D, FRC Path
Professor and Vice Chair Cell Biology and Physiology
Professor Immunology Director Center for Biologic Imaging
BSTS 225
University of Pittsburgh
3500 Terrace St
Pittsburgh PA 15261
412-352-2277
www.cbi.pitt.edu


-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Caroline Bass
Sent: Friday, January 14, 2011 11:10 AM
To: [hidden email]
Subject: Re: BSL3 microscopy

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Hi,

I've worked in a BSL3 lab before, and did use a general inverted
microscope. Are you sure that the specimen can't be contained and that you
need the face shield? The purpose of a BSL3 facility is to maintain
negative air pressure in case of an incident. A face shield protects
against accidental splashes but doesn't do anything for an aerosolized or
airborne agent (they easily get around a mask). So it seems if there is a
way to contain the specimen either in a sealed environment (flask, or
slide chamber) or to set up a shield between the user and the specimen,
that would serve the same purpose.

Also, what about a monitor situation? If you can contain the specimen from
the user then it seems your typical confocal setup, where most of the work
is done on a computer, contained within the BSL3 facility should be
sufficient.

Caroline Bass




On 1/14/11 9:32 AM, "Watkins, Simon C" <[hidden email]> wrote:

>*****
>To join, leave or search the confocal microscopy listserv, go to:
>http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>*****
>
>Folks, I am about to install an advanced imaging system (sweptfield
>confocal and widefield imaging combo) in a biosafety level 3 facility.
>To work in the facility one has to suit up and wear a rigid face mask, as
>the cells/pathogens will be on the scope and may be open to the
>environment.  This means that the eyepieces of the scope are essentially
>useless.  I wonder if any other listers have dealt with this problem and
>what their solution was? Obviously the widefield camera will help a lot,
>but it doesnt allow survey of the full field of view, as we are doing
>mostly flourescence a video camera isnt much use.... back in the day,
>there were some screen solutions....
>Looking for creative ideas
>S.
>
>Simon C. Watkins Ph.D, FRC Path
>Professor and Vice Chair Cell Biology and Physiology
>Professor Immunology Director Center for Biologic Imaging
>BSTS 225
>University of Pittsburgh
>3500 Terrace St
>Pittsburgh PA 15261
>412-352-2277
>www.cbi.pitt.edu<http://www.cbi.pitt.edu>

[S. Brunet]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Hello:

We are not BSL3, but we set up a camera + adapter at the eyepiece from Martin
Microscope (no financial affiliation) on our LSM410.  I can see many
fluorescent samples this way.
You could try a higher end camera and send the output to a monitor (some cameras
can be controlled using the computer software).
My biggest problem with the camera I have is the automatic focussing which can
correct for mis-focussing the objective (I can control that to an extent but I
prefer manual camera sometimes).
Vignetting will happen for a large field of view.  I can't remember off hand how
the field view compares to the eyepiece (probably a bit less).

I am not sure if this is the option you referred to in your post (that you say
would not work fo you).

I would assume that most new microscopes with a digital imaging camera would
suit your purpose... so I am trying to understand the problem...

Good luck,
Sophie
____________________________________________________
Sophie M. K. Brunet, Ph. D.
Research Officer
Optical Spectroscopy, Laser Systems and Applications
[hidden email]
306-966-1719 (office)   306-966-1702 (fax)
____________________________________________________
Saskatchewan Structural Sciences Centre
University of Saskatchewan
Thorvaldson Bldg.
110 Science Place
Saskatoon, Sk   S7N 5C9
____________________________________________________


Quoting "Watkins, Simon C" <[hidden email]>:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Folks, I am about to install an advanced imaging system (sweptfield confocal
> and widefield imaging combo) in a biosafety level 3 facility.  To work in the
> facility one has to suit up and wear a rigid face mask, as the
> cells/pathogens will be on the scope and may be open to the environment.
> This means that the eyepieces of the scope are essentially useless.  I wonder
> if any other listers have dealt with this problem and what their solution
> was? Obviously the widefield camera will help a lot, but it doesnt allow
> survey of the full field of view, as we are doing mostly flourescence a video
> camera isnt much use.... back in the day, there were some screen
> solutions....
> Looking for creative ideas
> S.
>
> Simon C. Watkins Ph.D, FRC Path
> Professor and Vice Chair Cell Biology and Physiology
> Professor Immunology Director Center for Biologic Imaging
> BSTS 225
> University of Pittsburgh
> 3500 Terrace St
> Pittsburgh PA 15261
> 412-352-2277
> www.cbi.pitt.edu<http://www.cbi.pitt.edu>
>

[Watkins, Simon C]

and did they work peter? (brightfield and fluorescence)
s

Simon C. Watkins Ph.D, FRC Path
Professor and Vice Chair Cell Biology and Physiology
Professor Immunology Director Center for Biologic Imaging
BSTS 225
University of Pittsburgh
3500 Terrace St
Pittsburgh PA 15261
412-352-2277
www.cbi.pitt.edu


-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Peter Humphreys
Sent: Friday, January 14, 2011 1:19 PM
To: [hidden email]
Subject: Re: BSL3 microscopy

We just demo'd some ivis pieces through Olympus uk if that helps. Peter
Sent from my HTC

----- Reply message -----
From: "Watkins, Simon C" <[hidden email]>
To: <[hidden email]>
Subject: BSL3 microscopy
Date: Fri, Jan 14, 2011 17:54


*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

This little question is proving quite fruitful for discussion, this is not a face shield but totally enclosed nonflexible mask with filtered air.  to stop any accidents aerosolized or otherwise, the bugs are right on the edge of a level 4 in my mind. we tried screens this am, frosted glass, not enough light.  video cameras not enough light, CCD cameras with a reducing adaptor will work but difficult and expensive.  The most interesting suggestion so far has been the vision engineering eyepieces.. They superficially look perfect http://www.visioneng.us/isis-ergonomic-microscope-eyepieces-overview.php but.... they stopped selling them 6 months ago... (still on the website, and they are hunting for some)


Simon C. Watkins Ph.D, FRC Path
Professor and Vice Chair Cell Biology and Physiology
Professor Immunology Director Center for Biologic Imaging
BSTS 225
University of Pittsburgh
3500 Terrace St
Pittsburgh PA 15261
412-352-2277
www.cbi.pitt.edu


-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Caroline Bass
Sent: Friday, January 14, 2011 11:10 AM
To: [hidden email]
Subject: Re: BSL3 microscopy

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Hi,

I've worked in a BSL3 lab before, and did use a general inverted
microscope. Are you sure that the specimen can't be contained and that you
need the face shield? The purpose of a BSL3 facility is to maintain
negative air pressure in case of an incident. A face shield protects
against accidental splashes but doesn't do anything for an aerosolized or
airborne agent (they easily get around a mask). So it seems if there is a
way to contain the specimen either in a sealed environment (flask, or
slide chamber) or to set up a shield between the user and the specimen,
that would serve the same purpose.

Also, what about a monitor situation? If you can contain the specimen from
the user then it seems your typical confocal setup, where most of the work
is done on a computer, contained within the BSL3 facility should be
sufficient.

Caroline Bass




On 1/14/11 9:32 AM, "Watkins, Simon C" <[hidden email]> wrote:

>*****
>To join, leave or search the confocal microscopy listserv, go to:
>http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>*****
>
>Folks, I am about to install an advanced imaging system (sweptfield
>confocal and widefield imaging combo) in a biosafety level 3 facility.
>To work in the facility one has to suit up and wear a rigid face mask, as
>the cells/pathogens will be on the scope and may be open to the
>environment.  This means that the eyepieces of the scope are essentially
>useless.  I wonder if any other listers have dealt with this problem and
>what their solution was? Obviously the widefield camera will help a lot,
>but it doesnt allow survey of the full field of view, as we are doing
>mostly flourescence a video camera isnt much use.... back in the day,
>there were some screen solutions....
>Looking for creative ideas
>S.
>
>Simon C. Watkins Ph.D, FRC Path
>Professor and Vice Chair Cell Biology and Physiology
>Professor Immunology Director Center for Biologic Imaging
>BSTS 225
>University of Pittsburgh
>3500 Terrace St
>Pittsburgh PA 15261
>412-352-2277
>www.cbi.pitt.edu<http://www.cbi.pitt.edu>

[leoncio vergara]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

If I understand well, the dilemma is that the camera in the side port would not let you see the complete field of view to efficiently search the sample as you can do with eye pieces... using the Confocal monitor to search the sample has the same problem of limited field of view and adds the problem of slow refresh rate and unnecessary photobleaching... plus is annoying...

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of S. Brunet
Sent: Friday, January 14, 2011 12:50 PM
To: [hidden email]
Subject: Re: BSL3 microscopy

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Hello:

We are not BSL3, but we set up a camera + adapter at the eyepiece from Martin
Microscope (no financial affiliation) on our LSM410.  I can see many
fluorescent samples this way.
You could try a higher end camera and send the output to a monitor (some cameras
can be controlled using the computer software).
My biggest problem with the camera I have is the automatic focussing which can
correct for mis-focussing the objective (I can control that to an extent but I
prefer manual camera sometimes).
Vignetting will happen for a large field of view.  I can't remember off hand how
the field view compares to the eyepiece (probably a bit less).

I am not sure if this is the option you referred to in your post (that you say
would not work fo you).

I would assume that most new microscopes with a digital imaging camera would
suit your purpose... so I am trying to understand the problem...

Good luck,
Sophie
____________________________________________________
Sophie M. K. Brunet, Ph. D.
Research Officer
Optical Spectroscopy, Laser Systems and Applications
[hidden email]
306-966-1719 (office)   306-966-1702 (fax)
____________________________________________________
Saskatchewan Structural Sciences Centre
University of Saskatchewan
Thorvaldson Bldg.
110 Science Place
Saskatoon, Sk   S7N 5C9
____________________________________________________


Quoting "Watkins, Simon C" <[hidden email]>:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Folks, I am about to install an advanced imaging system (sweptfield confocal
> and widefield imaging combo) in a biosafety level 3 facility.  To work in the
> facility one has to suit up and wear a rigid face mask, as the
> cells/pathogens will be on the scope and may be open to the environment.
> This means that the eyepieces of the scope are essentially useless.  I wonder
> if any other listers have dealt with this problem and what their solution
> was? Obviously the widefield camera will help a lot, but it doesnt allow
> survey of the full field of view, as we are doing mostly flourescence a video
> camera isnt much use.... back in the day, there were some screen
> solutions....
> Looking for creative ideas
> S.
>
> Simon C. Watkins Ph.D, FRC Path
> Professor and Vice Chair Cell Biology and Physiology
> Professor Immunology Director Center for Biologic Imaging
> BSTS 225
> University of Pittsburgh
> 3500 Terrace St
> Pittsburgh PA 15261
> 412-352-2277
> www.cbi.pitt.edu<http://www.cbi.pitt.edu>
>

[Craig Brideau]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

The field of view depends on the optics used to couple the camera to the
microscope, and the size of the CCD array.  CCD arrays are square, and the
image from a microscope is round.  To prevent unsightly black corners from
appearing in your image they usually overfill the CCD array.  That is, the
CCD is only imaging a portion of the full field of view to deal with the
circle vs rectangle problem.  If you are willing to accept not using the
whole CCD array and throw out those black corners, you can set the optics to
only image the specimen plane onto the middle part of the CCD.  This will
give you the full field of view the microscope is capable of, at a cost of
some resolution.  If you are just trying to locate your samples in
preparation for confocal imaging though, this shouldn't be a big deal.

Craig



On Fri, Jan 14, 2011 at 12:14 PM, Vergara, Leoncio A. <[hidden email]>wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> If I understand well, the dilemma is that the camera in the side port would
> not let you see the complete field of view to efficiently search the sample
> as you can do with eye pieces... using the Confocal monitor to search the
> sample has the same problem of limited field of view and adds the problem of
> slow refresh rate and unnecessary photobleaching... plus is annoying...
>
> -----Original Message-----
> From: Confocal Microscopy List [mailto:[hidden email]]
> On Behalf Of S. Brunet
> Sent: Friday, January 14, 2011 12:50 PM
> To: [hidden email]
> Subject: Re: BSL3 microscopy
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hello:
>
> We are not BSL3, but we set up a camera + adapter at the eyepiece from
> Martin
> Microscope (no financial affiliation) on our LSM410.  I can see many
> fluorescent samples this way.
> You could try a higher end camera and send the output to a monitor (some
> cameras
> can be controlled using the computer software).
> My biggest problem with the camera I have is the automatic focussing which
> can
> correct for mis-focussing the objective (I can control that to an extent
> but I
> prefer manual camera sometimes).
> Vignetting will happen for a large field of view.  I can't remember off
> hand how
> the field view compares to the eyepiece (probably a bit less).
>
> I am not sure if this is the option you referred to in your post (that you
> say
> would not work fo you).
>
> I would assume that most new microscopes with a digital imaging camera
> would
> suit your purpose... so I am trying to understand the problem...
>
> Good luck,
> Sophie
> ____________________________________________________
> Sophie M. K. Brunet, Ph. D.
> Research Officer
> Optical Spectroscopy, Laser Systems and Applications
> [hidden email]
> 306-966-1719 (office)   306-966-1702 (fax)
> ____________________________________________________
> Saskatchewan Structural Sciences Centre
> University of Saskatchewan
> Thorvaldson Bldg.
> 110 Science Place
> Saskatoon, Sk   S7N 5C9
> ____________________________________________________
>
>
> Quoting "Watkins, Simon C" <[hidden email]>:
>
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > *****
> >
> > Folks, I am about to install an advanced imaging system (sweptfield
> confocal
> > and widefield imaging combo) in a biosafety level 3 facility.  To work in
> the
> > facility one has to suit up and wear a rigid face mask, as the
> > cells/pathogens will be on the scope and may be open to the environment.
> > This means that the eyepieces of the scope are essentially useless.  I
> wonder
> > if any other listers have dealt with this problem and what their solution
> > was? Obviously the widefield camera will help a lot, but it doesnt allow
> > survey of the full field of view, as we are doing mostly flourescence a
> video
> > camera isnt much use.... back in the day, there were some screen
> > solutions....
> > Looking for creative ideas
> > S.
> >
> > Simon C. Watkins Ph.D, FRC Path
> > Professor and Vice Chair Cell Biology and Physiology
> > Professor Immunology Director Center for Biologic Imaging
> > BSTS 225
> > University of Pittsburgh
> > 3500 Terrace St
> > Pittsburgh PA 15261
> > 412-352-2277
> > www.cbi.pitt.edu<http://www.cbi.pitt.edu>
> >
>

[James Pawley]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Hi all,

Up until recently, it was hard to improve on the dark-adapted human
eye as a detector for live-time widefield LM images.

Other image sensors might be better able to integrate weak signals
over time or to perform well over a limited field of view, but in
real-time, the rotatable eyeball was really hard to beat. Until last
year the closest challenger was the EM-CCD, but this had relatively
few pixels, only moderate readout speed and no color.

It is my impression that the new scientific CMOS detectors (such as
those recently offered by Hamamatsu and Andor, no connection) may
soon overcome these limitations. While it is true that, so far, they
do not provide color images, they do have a wide field of view as
well as fast readout, very low read noise and quantum efficiency
significantly higher than that of the human eye.

I see no reason why, in the future, they could not be coated with
color filters to produce a single-chip color sensor. Although the
color filters will reduce the QE to white light by a factor of 3 or
4, it is also true that the cones which allow human eyes to sense
color are far less sensitive than the monochrome rods. So I think
that, as long as we can figure out how to handle the massive
information flow associated with sorting and displaying 5Mpixel,
3-color, at video rates, we will still come out ahead.

Such image sensors may eventually replace the oculars and prism heads
now common on scientific microscopes. Just compare the cost of these
optics with that of a potentially mass-produced chip. Not only this,
but such a system greatly facilitates keeping the specimen in the
dark while the operator works in ambient light.

Indeed, we already see several manufactures (such as AMG, Mill Creek
Washington. No connection) offering fluorescence microscopes in which
all imaging is carried out via CCD sensors and LCD displays.

Until we come up with a "jack" to send such images directly into the
optical centers of the brain (Heaven Forbid!), this may be the best
we can do.

Cheers,

Jim P.

NOTE: I have no connection with Hamamatsu, AMG or Andor except that,
like virtually all other microscope manufacturer, they have in the
past provided instruments for use by students use at the UBC
Live-Cell Course. I mention these companies only as examples that I
happen to know about but I do so in the awareness that many other
companies will soon be applying these and other sCMOS sensors in much
the same way.
***************************************************************************
Prof. James B. Pawley,                          Ph.
608-238-3953                        
21. N. Prospect Ave. Madison, WI 53726 USA
[hidden email]
3D Microscopy of Living Cells Course, June 11-23, 2011, UBC, Vancouver Canada
Info: http://www.3dcourse.ubc.ca/            Applications due by March 15, 2011
               "If it ain't diffraction, it must be statistics." Anon.

>*****
>To join, leave or search the confocal microscopy listserv, go to:
>http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>*****
>
>Good afternoon,
>
>the straight forward way, if an additional camera port is available on the
>microscope, might be to mount a CCD camera and between the microscope and
>the CCD camera a C-mount adapter including a 0.32x or 0.5x scaler. The
>disadvantage is that the image produced by the camera will show "black
>corners" around the image of the FOV since the entire FOV is projected
>onto the CCD chip, which is rectangular, while the FOV is cicular in
>shape.
>
>If you canNOT proceed this way:
>You might try what just is an idea and I do not know whether this would work:
>
>The eyepiece, as many of the group members know, is used as a
>magnification glass to observe the real intermediate image, which, in
>most, possibly all, standards is located 10mm inside the tube which holds
>the eyepiece(s), distance measured from the edge to which the eyepiece is
>set.
>
>If your preps produce images with sufficiently large contrasts (otherwise
>use DIC or PC or another contrast method):
>If you would place (a) circular screen(s) made from a whitish,
>structureless material or possibly frozen glass into the tube(s) holding
>the eyepiece(s) so that the screens are positioned 10mm inside the
>tube(s), you would be able to view the real intermediate on these small
>screens. Of course, the image is much too small for eye observation, but
>you might place a digital photoapparatus in "near view mode" so that it
>will image this real intermediate image. I've just tried with my Olympus
>my 810, and it easily images the 25mm which is, as far as I know, the
>largest diameter of the FOV in the plane of the real intermediate image.
>
>
>Best wishes,
>
>Johannes
>
>
>>  *****
>>  To join, leave or search the confocal microscopy listserv, go to:
>>  http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>>  *****
>>
>>  Folks, I am about to install an advanced imaging system (sweptfield
>>  confocal and widefield imaging combo) in a biosafety level 3 facility.  To
>>  work in the facility one has to suit up and wear a rigid face mask, as the
>>  cells/pathogens will be on the scope and may be open to the environment.
>>  This means that the eyepieces of the scope are essentially useless.  I
>>  wonder if any other listers have dealt with this problem and what their
>>  solution was? Obviously the widefield camera will help a lot, but it
>>  doesnt allow survey of the full field of view, as we are doing mostly
>>  flourescence a video camera isnt much use.... back in the day, there were
>>  some screen solutions....
>>  Looking for creative ideas
>>  S.
>>
>>  Simon C. Watkins Ph.D, FRC Path
>>  Professor and Vice Chair Cell Biology and Physiology
>>  Professor Immunology Director Center for Biologic Imaging
>>  BSTS 225
>>  University of Pittsburgh
>>  3500 Terrace St
>>  Pittsburgh PA 15261
>>  412-352-2277
>>  www.cbi.pitt.edu<http://www.cbi.pitt.edu>
>>
>
>
>--
>P. Johannes Helm
>
>Voice: ([phone] (office)
>Fax: [phone] (office)

--
***************************************************************************
Prof. James B. Pawley,                          Ph.
608-238-3953                        
21. N. Prospect Ave. Madison, WI 53726 USA
[hidden email]
3D Microscopy of Living Cells Course, June 11-23, 2011, UBC, Vancouver Canada
Info: http://www.3dcourse.ubc.ca/            Applications due by March 15, 2011
               "If it ain't diffraction, it must be statistics." Anon.

[Peter Humphreys]

Brightfield was great,did not get opportunity to try fluorescence,we are buying for stereo scope in a cat2 cabinet.
But for fluorescence they are of interest to us also.
Peter

----- Reply message -----
From: "Watkins, Simon C" <[hidden email]>
To: <[hidden email]>
Subject: BSL3 microscopy
Date: Fri, Jan 14, 2011 18:50


and did they work peter? (brightfield and fluorescence)
s

Simon C. Watkins Ph.D, FRC Path
Professor and Vice Chair Cell Biology and Physiology
Professor Immunology Director Center for Biologic Imaging
BSTS 225
University of Pittsburgh
3500 Terrace St
Pittsburgh PA 15261
412-352-2277
www.cbi.pitt.edu


-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Peter Humphreys
Sent: Friday, January 14, 2011 1:19 PM
To: [hidden email]
Subject: Re: BSL3 microscopy

We just demo'd some ivis pieces through Olympus uk if that helps. Peter
Sent from my HTC

----- Reply message -----
From: "Watkins, Simon C" <[hidden email]>
To: <[hidden email]>
Subject: BSL3 microscopy
Date: Fri, Jan 14, 2011 17:54


*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

This little question is proving quite fruitful for discussion, this is not a face shield but totally enclosed nonflexible mask with filtered air.  to stop any accidents aerosolized or otherwise, the bugs are right on the edge of a level 4 in my mind. we tried screens this am, frosted glass, not enough light.  video cameras not enough light, CCD cameras with a reducing adaptor will work but difficult and expensive.  The most interesting suggestion so far has been the vision engineering eyepieces.. They superficially look perfect http://www.visioneng.us/isis-ergonomic-microscope-eyepieces-overview.php but.... they stopped selling them 6 months ago... (still on the website, and they are hunting for some)


Simon C. Watkins Ph.D, FRC Path
Professor and Vice Chair Cell Biology and Physiology
Professor Immunology Director Center for Biologic Imaging
BSTS 225
University of Pittsburgh
3500 Terrace St
Pittsburgh PA 15261
412-352-2277
www.cbi.pitt.edu


-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Caroline Bass
Sent: Friday, January 14, 2011 11:10 AM
To: [hidden email]
Subject: Re: BSL3 microscopy

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Hi,

I've worked in a BSL3 lab before, and did use a general inverted
microscope. Are you sure that the specimen can't be contained and that you
need the face shield? The purpose of a BSL3 facility is to maintain
negative air pressure in case of an incident. A face shield protects
against accidental splashes but doesn't do anything for an aerosolized or
airborne agent (they easily get around a mask). So it seems if there is a
way to contain the specimen either in a sealed environment (flask, or
slide chamber) or to set up a shield between the user and the specimen,
that would serve the same purpose.

Also, what about a monitor situation? If you can contain the specimen from
the user then it seems your typical confocal setup, where most of the work
is done on a computer, contained within the BSL3 facility should be
sufficient.

Caroline Bass




On 1/14/11 9:32 AM, "Watkins, Simon C" <[hidden email]> wrote:

>*****
>To join, leave or search the confocal microscopy listserv, go to:
>http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>*****
>
>Folks, I am about to install an advanced imaging system (sweptfield
>confocal and widefield imaging combo) in a biosafety level 3 facility.
>To work in the facility one has to suit up and wear a rigid face mask, as
>the cells/pathogens will be on the scope and may be open to the
>environment.  This means that the eyepieces of the scope are essentially
>useless.  I wonder if any other listers have dealt with this problem and
>what their solution was? Obviously the widefield camera will help a lot,
>but it doesnt allow survey of the full field of view, as we are doing
>mostly flourescence a video camera isnt much use.... back in the day,
>there were some screen solutions....
>Looking for creative ideas
>S.
>
>Simon C. Watkins Ph.D, FRC Path
>Professor and Vice Chair Cell Biology and Physiology
>Professor Immunology Director Center for Biologic Imaging
>BSTS 225
>University of Pittsburgh
>3500 Terrace St
>Pittsburgh PA 15261
>412-352-2277
>www.cbi.pitt.edu<http://www.cbi.pitt.edu>

[Rosemary.White]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Hi Simon,

Alex Hyatt has installed a confocal in their PC4 facility within AAHL in
Geelong, just south of Melbourne.
website: http://www.csiro.au/services/ABMF.html

cheers,
Rosemary

Dr Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
E [hidden email]


On 15/01/11 1:32 AM, "Watkins, Simon C" <[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Folks, I am about to install an advanced imaging system (sweptfield confocal
> and widefield imaging combo) in a biosafety level 3 facility.  To work in the
> facility one has to suit up and wear a rigid face mask, as the cells/pathogens
> will be on the scope and may be open to the environment.  This means that the
> eyepieces of the scope are essentially useless.  I wonder if any other listers
> have dealt with this problem and what their solution was? Obviously the
> widefield camera will help a lot, but it doesnt allow survey of the full field
> of view, as we are doing mostly flourescence a video camera isnt much use....
> back in the day, there were some screen solutions....
> Looking for creative ideas
> S.
>
> Simon C. Watkins Ph.D, FRC Path
> Professor and Vice Chair Cell Biology and Physiology
> Professor Immunology Director Center for Biologic Imaging
> BSTS 225
> University of Pittsburgh
> 3500 Terrace St
> Pittsburgh PA 15261
> 412-352-2277
> www.cbi.pitt.edu<http://www.cbi.pitt.edu>

[George McNamara]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Hi Simon,

Leave the eyepieces in the drawer and teach your users to use the live
camera images. Would also be useful to have an identical microscope in
the main lab that users can learn on and adapt to.

You can also take a tip from Sedat lab's and Applied Precision's use of
a "scout scope". More practically, in your BSL3 system, low
magnification lens and/or motorized XY(Z) tile scanning.

You could take this even further by having a duplicate computer monitor,
keyboard, mouse, etc, outside the BSL3 space. Preferably with the
microscope on the other side of a window from the non-3 station. Same
ideas would apply to a microscope in a GMP facility (which is the idea
came from).

George

p.s. Are you really studying flour in there? ... flourescence

On 1/14/2011 9:32 AM, Watkins, Simon C wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Folks, I am about to install an advanced imaging system (sweptfield confocal and widefield imaging combo) in a biosafety level 3 facility.  To work in the facility one has to suit up and wear a rigid face mask, as the cells/pathogens will be on the scope and may be open to the environment.  This means that the eyepieces of the scope are essentially useless.  I wonder if any other listers have dealt with this problem and what their solution was? Obviously the widefield camera will help a lot, but it doesnt allow survey of the full field of view, as we are doing mostly flourescence a video camera isnt much use.... back in the day, there were some screen solutions....
> Looking for creative ideas
> S.
>
> Simon C. Watkins Ph.D, FRC Path
> Professor and Vice Chair Cell Biology and Physiology
> Professor Immunology Director Center for Biologic Imaging
> BSTS 225
> University of Pittsburgh
> 3500 Terrace St
> Pittsburgh PA 15261
> 412-352-2277
> www.cbi.pitt.edu<http://www.cbi.pitt.edu>
>
>