fluorescence microscope as plate reader

[Christoph Heyder]

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Hi all,
perhaps this is the wrong forum, but I would like to know if it is
possible to use an inverted widefield fluorescence microscope (with a
motorized xy stage and CCD-camera) as a fluorescent plate reader, e.g.
for assessing fluorescence intensity or counting cells at the bottom of
the plate. I  have googled but so far have found nothing. Any
suggestions are welcome.
Sincerely,
Chris

[Yan Gu]

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Dear Chris,

We have done this. We used IX70 from Olympus, Prior stage and software from
Objective Imaging. Check either Objective Imaging or Cybermedia UK for more
details.


Best wishes

Yan

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On
Behalf Of Christoph Heyder
Sent: 11 February 2013 09:49
To: [hidden email]
Subject: fluorescence microscope as plate reader

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Hi all,
perhaps this is the wrong forum, but I would like to know if it is possible
to use an inverted widefield fluorescence microscope (with a motorized xy
stage and CCD-camera) as a fluorescent plate reader, e.g.
for assessing fluorescence intensity or counting cells at the bottom of the
plate. I  have googled but so far have found nothing. Any suggestions are
welcome.
Sincerely,
Chris

[Csúcs Gábor-2]

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Dear Chris,

I see no problem with your application - many microscope control software
allows such acquisition (commercial but also MicroManager) and also for
the analysis you'll find both commercial and opensource solutions (like
CellProfiler or ImageJ).

Greetings    Gabor





On 2/11/13 10:49 AM, "Christoph Heyder"
<[hidden email]> wrote:

>*****
>To join, leave or search the confocal microscopy listserv, go to:
>http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>*****
>
>Hi all,
>perhaps this is the wrong forum, but I would like to know if it is
>possible to use an inverted widefield fluorescence microscope (with a
>motorized xy stage and CCD-camera) as a fluorescent plate reader, e.g.
>for assessing fluorescence intensity or counting cells at the bottom of
>the plate. I  have googled but so far have found nothing. Any
>suggestions are welcome.
>Sincerely,
>Chris

[mcammer]

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The microscopy forum http://www.microscopy.com/ is probably more the place to ask, but here is ok.

The short answer is yes.  A longer answer depends on your specific equipment and software.  If you have a electronically controlled XY stage and shutters/filter wheels, there are a lot of software packages that handle this including free MicroManager.  See http://valelab.ucsf.edu/~MM/MMwiki/ 

________________________________________________________
Michael Cammer, Assistant Research Scientist
Skirball Institute of Biomolecular Medicine
Lab: [phone]  Cell: [phone]


-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Christoph Heyder
Sent: Monday, February 11, 2013 4:49 AM
To: [hidden email]
Subject: fluorescence microscope as plate reader

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

Hi all,
perhaps this is the wrong forum, but I would like to know if it is possible to use an inverted widefield fluorescence microscope (with a motorized xy stage and CCD-camera) as a fluorescent plate reader, e.g.
for assessing fluorescence intensity or counting cells at the bottom of the plate. I  have googled but so far have found nothing. Any suggestions are welcome.
Sincerely,
Chris

[Barbara Foster]

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Hi, Chris

You can certainly do this... there are stage holders which will hold
plates of different sizes and configurations.

The two issues you need to be aware of are the hysteresis in stage
movement (fixed by moving more slowly and in one direction rather
than a zig zag) and optical effects at the edge of the wells where
the transition from the bottom to the side of a well causes a
"lensing" effect.  Depending on the relative size of cells to well,
you can ofter counter this effect by changing your counting rules and
counting in a smaller area.  Also, the more on-axis the viewing the better.

Hope this is helpful.

Best regards,
Barbara Foster, President & Chief Consultant
Microscopy/Microscopy Education*
www.MicroscopyEducation.com

*A subsidiary of The Microscopy & Imaging Place, Inc.
7101 Royal Glen Trail, Suite A
McKinney, TX 75070
P: 972-924-5310
F: 214-592-0277

MME is currently scheduling courses for the Fall 2013. Call us today
for a free training evaluation.


At 09:25 PM 2/10/2013, Christoph Heyder wrote:

>*****
>To join, leave or search the confocal microscopy listserv, go to:
>http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>*****
>
>Hi all,
>perhaps this is the wrong forum, but I would like to know if it is
>possible to use an inverted widefield fluorescence microscope (with
>a motorized xy stage and CCD-camera) as a fluorescent plate reader,
>e.g. for assessing fluorescence intensity or counting cells at the
>bottom of the plate. I  have googled but so far have found nothing.
>Any suggestions are welcome.
>Sincerely,
>Chris

[mahogny]

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I strongly suggest micro-manager, as in the previous post. It works on all
operating systems and if you ever need to do something more fancy than just
reading fluorescence, then the system can cover that as well, and you won't
have to think of buying *yet another* software which tends to be the case
for commercial offerings.

/Johan


On Mon, Feb 11, 2013 at 3:59 PM, Cammer, Michael <[hidden email]
> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> The microscopy forum http://www.microscopy.com/ is probably more the
> place to ask, but here is ok.
>
> The short answer is yes.  A longer answer depends on your specific
> equipment and software.  If you have a electronically controlled XY stage
> and shutters/filter wheels, there are a lot of software packages that
> handle this including free MicroManager.  See
> http://valelab.ucsf.edu/~MM/MMwiki/
>
> ________________________________________________________
> Michael Cammer, Assistant Research Scientist
> Skirball Institute of Biomolecular Medicine
> Lab: [phone]  Cell: [phone]
>
>
> -----Original Message-----
> From: Confocal Microscopy List [mailto:[hidden email]]
> On Behalf Of Christoph Heyder
> Sent: Monday, February 11, 2013 4:49 AM
> To: [hidden email]
> Subject: fluorescence microscope as plate reader
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hi all,
> perhaps this is the wrong forum, but I would like to know if it is
> possible to use an inverted widefield fluorescence microscope (with a
> motorized xy stage and CCD-camera) as a fluorescent plate reader, e.g.
> for assessing fluorescence intensity or counting cells at the bottom of
> the plate. I  have googled but so far have found nothing. Any suggestions
> are welcome.
> Sincerely,
> Chris
>

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Karolinska Institutet
Ecobima AB - Custom solutions for life sciences
http://www.ecobima.com  http://mahogny.areta.org  http://www.endrov.net

<http://www.endrov.net>