Objective for visual fluorescence inspection in plastic plates

[lechristophe]

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Dear all,

I'd like to get the brightest possible air objective for visual
checking of transfected cell cultures by epifluorescence (live cells
or immunolabeled).

The cells arte on 0.15 mm glass coverlips inside 12-well cell culture
plates, and I wonder what working distance would be needed. What is
the typical thickness of the plastic bottom of these plates ? Would a
1.5 - 2mm working distance objective (such as a NA 0.5 10X with 1.9 mm
or a NA0.5 20X with 2.1 mm) work, or should I aim for a > 4mm WD
objective that have lower NA (<0.3) ?


Thanks for your help,

Christophe

--
Christophe Leterrier
Researcher
Axonal Domains Architecture Team
CRN2M CNRS UMR 7286
Aix Marseille University, France

[mmodel]

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We often use x10/0.4 planapo. Not very scientific but works for GFP transfection

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Christophe Leterrier
Sent: Wednesday, November 21, 2012 10:41 AM
To: [hidden email]
Subject: Objective for visual fluorescence inspection in plastic plates

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Dear all,

I'd like to get the brightest possible air objective for visual
checking of transfected cell cultures by epifluorescence (live cells
or immunolabeled).

The cells arte on 0.15 mm glass coverlips inside 12-well cell culture
plates, and I wonder what working distance would be needed. What is
the typical thickness of the plastic bottom of these plates ? Would a
1.5 - 2mm working distance objective (such as a NA 0.5 10X with 1.9 mm
or a NA0.5 20X with 2.1 mm) work, or should I aim for a > 4mm WD
objective that have lower NA (<0.3) ?


Thanks for your help,

Christophe

--
Christophe Leterrier
Researcher
Axonal Domains Architecture Team
CRN2M CNRS UMR 7286
Aix Marseille University, France

[Glen MacDonald-2]

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A 10x achromat with the higher NA  might be brightest so long as depth of field brings the larger field of view into acceptable focus.  

Regards,
glen
Glen MacDonald
Core for Communication Research
Virginia Merrill Bloedel Hearing Research Center
Box 357923
University of Washington
Seattle, WA 98195-7923  USA
[phone]
[hidden email]


On Nov 21, 2012, at 7:41 AM, Christophe Leterrier wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Dear all,
>
> I'd like to get the brightest possible air objective for visual
> checking of transfected cell cultures by epifluorescence (live cells
> or immunolabeled).
>
> The cells arte on 0.15 mm glass coverlips inside 12-well cell culture
> plates, and I wonder what working distance would be needed. What is
> the typical thickness of the plastic bottom of these plates ? Would a
> 1.5 - 2mm working distance objective (such as a NA 0.5 10X with 1.9 mm
> or a NA0.5 20X with 2.1 mm) work, or should I aim for a > 4mm WD
> objective that have lower NA (<0.3) ?
>
>
> Thanks for your help,
>
> Christophe
>
> --
> Christophe Leterrier
> Researcher
> Axonal Domains Architecture Team
> CRN2M CNRS UMR 7286
> Aix Marseille University, France

[Craig Brideau]

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Olympus makes a 20x culture flask lens that gives decent performance.  We
use it for looking through ~2mm thick plastic walls and the occasional
thick glass plate to examine cells.  It has a correction collar so you can
tune it for thickness.

Craig


On Wed, Nov 21, 2012 at 9:52 AM, Glen MacDonald <[hidden email]>wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> A 10x achromat with the higher NA  might be brightest so long as depth of
> field brings the larger field of view into acceptable focus.
>
> Regards,
> glen
> Glen MacDonald
> Core for Communication Research
> Virginia Merrill Bloedel Hearing Research Center
> Box 357923
> University of Washington
> Seattle, WA 98195-7923  USA
> [phone]
> [hidden email]
>
>
> On Nov 21, 2012, at 7:41 AM, Christophe Leterrier wrote:
>
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > *****
> >
> > Dear all,
> >
> > I'd like to get the brightest possible air objective for visual
> > checking of transfected cell cultures by epifluorescence (live cells
> > or immunolabeled).
> >
> > The cells arte on 0.15 mm glass coverlips inside 12-well cell culture
> > plates, and I wonder what working distance would be needed. What is
> > the typical thickness of the plastic bottom of these plates ? Would a
> > 1.5 - 2mm working distance objective (such as a NA 0.5 10X with 1.9 mm
> > or a NA0.5 20X with 2.1 mm) work, or should I aim for a > 4mm WD
> > objective that have lower NA (<0.3) ?
> >
> >
> > Thanks for your help,
> >
> > Christophe
> >
> > --
> > Christophe Leterrier
> > Researcher
> > Axonal Domains Architecture Team
> > CRN2M CNRS UMR 7286
> > Aix Marseille University, France
>