Imaging via the microscope condenser

[Csúcs Gábor-3]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Dear All,

On laser scanning confocals it is an often used imaging method to record a transmitted image via the condenser. My question is whether anyone of you used the same approach to record a "normal" wide-field transmitted image (e.g. parallel to the fluorescent one). If yes, could you please share your experience?

Thanks        Gabor

[jerie]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Hi Gábor,

Electro physiologists use Dodt contrast imaging for patching on their
scanning microscopes,  for example
http://m.sciencemag.org/site/feature/data/1042873.xhtml

No commercial interest.

Cheers,  Jens

Visiting Scientist
Fundação Oswaldo Cruz - Ministério da Saúde, Centro de Desenvolvimento
Tecnológico em Saúde (CDTS)
Am 23.06.2015 07:28 schrieb "Csúcs Gábor" <[hidden email]>:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Dear All,
>
> On laser scanning confocals it is an often used imaging method to record a
> transmitted image via the condenser. My question is whether anyone of you
> used the same approach to record a "normal" wide-field transmitted image
> (e.g. parallel to the fluorescent one). If yes, could you please share your
> experience?
>
> Thanks        Gabor
>

[George McNamara]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Hi again Gabor,

Yes, the CompuCyte laser scanning cytometer (LSC) and iCys and iCyte
imaging scanning cytometers, now part of ThorLabs,
https://www.thorlabs.com/newgrouppage9.cfm?objectgroup_id=7473
https://www.thorlabs.com/newgrouppage9.cfm?objectgroup_id=7676

routinely acquired transmitted light data through the condenser, and in
the case of the "i" instruments, produced an image.

The Huron Digital Patyhology (previously Huron Technologies, and
previous to that Biomedical Photometrics Inc) TissueScope product line
is typically both brightfield and confocal fluorescence. Many of the
TissueScope products can use a large stage enabling scanning an entire
adult human brain slice.
http://www.hurondigitalpathology.com/solutions/tissuescope-cf/

Several research groups have replaced the condenser lens (which is often
as cheap a lens as the big four can get away with) with an objective
lens. I first saw this in the mid-1980s by David Soll and his group at
University of Iowa - to enable tracking live cell morphology details in
higher time resolution by imaging both up and down.

More recently, 4pi, 4pi-STED, and iPALM are dual objective lenses facing
each other. Some SPIM/DSLM systems also have face-to-face lenses, along
with a third, and (if I recall correctly) in some configurations fourth,
perpendicular.

Enjoy,

George


On 6/23/2015 5:27 AM, Csúcs  Gábor wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Dear All,
>
> On laser scanning confocals it is an often used imaging method to record a transmitted image via the condenser. My question is whether anyone of you used the same approach to record a "normal" wide-field transmitted image (e.g. parallel to the fluorescent one). If yes, could you please share your experience?
>
> Thanks        Gabor
>
>    

--



George McNamara, Ph.D.
Single Cells Analyst
L.J.N. Cooper Lab
University of Texas M.D. Anderson Cancer Center
Houston, TX 77054
Tattletales http://works.bepress.com/gmcnamara/42

[James Kerin]

Dear Gabor,
We routinely do this for widefield imaging or, photometry (using a photomultiplier to record fluorescence from an optically defined ROI). &nbsp;If I understand your question correctly then you need a red or infrared filter in line with the transmitted light source and a second camera port, filter wheel or image splitter device so as to record the red/ir transmitted alongside the fluorescence wavelengths. Typical configurations would be:&nbsp;
(a) simultaneous transmitted/fluorecence using a dichroic beamsplitter to divert light between sensitive (fluorescence) and cheap (transmitted) cameras(b) sequentially with fluorescence using an automated filter wheel to change emitters(c) in parallel with fluorescence on your imaging camera using one channel of an image splitter for the transmitted light and the other(s) for fluorescence
The main thing is to choose a transmitted wavelength that doesn't hurt your cells or overlap with the required fluorescence; and that will pass through the microscope dichroic. &nbsp;We have a range of dedicated products to achieve the configurations above and would be very happy to advise further

 
 
 




J. P.  Kerin
Marketing Director
Cairn Research  Ltd
Graveney Road
Faversham
Kent, ME13 8UP
UK


Tel: + 44 (0)1795 594507
&nbsp;&nbsp;filters and mirrors now in stock!


&nbsp;




On  06/23/2015, 11:27am, Cs&uacute;cs  G&aacute;bor ([hidden email]) wrote:
*****
 
To join, leave or search the confocal microscopy listserv, go to:
 
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
 
Post images on http://www.imgur.com and include the link in your posting.
 
*****
 

 
Dear All,
 

 
On laser scanning confocals it is an often used imaging method to record a transmitted image via the condenser. My question is whether anyone of you used the same approach to record a "normal" wide-field transmitted image (e.g. parallel to the fluorescent one). If yes, could you please share your experience?
 

 
Thanks        Gabor

[Craig Brideau]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

I constructed a DODT system a number of years back using an oil-immersion
condenser. As long as you have a high NA condenser you can do all kinds of
imaging with it. We used a 785nm LED array for illumination and a similar
band-pass filter in front of the camera for imaging. It was a very easy
modification since many systems already include a 785nm filter for the
camera and the LED and DODT gradient generator can be attached to any ready
lamp port.

Craig

On Tue, Jun 23, 2015 at 7:23 AM, James Kerin <[hidden email]>
wrote:

> Dear Gabor,
> We routinely do this for widefield imaging or, photometry (using a
> photomultiplier to record fluorescence from an optically defined ROI).
> &nbsp;If I understand your question correctly then you need a red or
> infrared filter in line with the transmitted light source and a second
> camera port, filter wheel or image splitter device so as to record the
> red/ir transmitted alongside the fluorescence wavelengths. Typical
> configurations would be:&nbsp;
> (a) simultaneous transmitted/fluorecence using a dichroic beamsplitter to
> divert light between sensitive (fluorescence) and cheap (transmitted)
> cameras(b) sequentially with fluorescence using an automated filter wheel
> to change emitters(c) in parallel with fluorescence on your imaging camera
> using one channel of an image splitter for the transmitted light and the
> other(s) for fluorescence
> The main thing is to choose a transmitted wavelength that doesn't hurt
> your cells or overlap with the required fluorescence; and that will pass
> through the microscope dichroic. &nbsp;We have a range of dedicated
> products to achieve the configurations above and would be very happy to
> advise further
>
>
>
>
>
>
>
>
> J. P.  Kerin
> Marketing Director
> Cairn Research  Ltd
> Graveney Road
> Faversham
> Kent, ME13 8UP
> UK
>
>
> Tel: + 44 (0)1795 594507
> &nbsp;&nbsp;filters and mirrors now in stock!
>
>
> &nbsp;
>
>
>
>
> On  06/23/2015, 11:27am, Cs&uacute;cs  G&aacute;bor (
> [hidden email]) wrote:
> *****
>
> To join, leave or search the confocal microscopy listserv, go to:
>
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>
> Post images on http://www.imgur.com and include the link in your posting.
>
> *****
>
>
>
> Dear All,
>
>
>
> On laser scanning confocals it is an often used imaging method to record a
> transmitted image via the condenser. My question is whether anyone of you
> used the same approach to record a "normal" wide-field transmitted image
> (e.g. parallel to the fluorescent one). If yes, could you please share your
> experience?
>
>
>
> Thanks        Gabor
>