*****
To join or leave the confocal microscopy listserv or to change your email address, go to:
https://lists.umn.edu/cgi-bin/wa?SUBED1=confocalmicroscopy&A=1
Post images on http://www.imgur.com and include the link in your posting.
*****

Hi Emmanuel,

three seconds is a lot, I think it takes less than 0.5 s on our Ti-E to
change a filter cube, making it less than a second per image set. You may
be able to optimize this by changing the order of cubes in the turret.

As Paula mentioned, a polarizer in the emission filter wheel will be much
faster. But note that the spinning disk itself will "eat" 99% of the light
(that's it's job, after all), so you may not get enough light for a decent
DIC image with not-too-long exposure. Also, the polarization may be messed
up by the W1 optics (like the main dichroic), so you may end up with
less-than-optimal DIC contrast (a green filter in the illumination column
might help, but I haven't tested this).

To get the best DIC we use the "Bypass" mode of the W-1 (I'm not sure if
this is available in all W-1 units) - that bypasses the disk and allows all
the light to reach the camera. But that requires another mechanical part to
move, again taking about 1/2 second or so.

Another option would be to engage the polarizer (in the filter turret) for
both fluorescence and DIC, but that will cost you >50% (actually over 60%)
of the emitted fluorescence intensity, and you'll have to increase the
laser power too, in order to get the same intensity at the sample
(depending on how well and in what direction your lasers are polarized).

In the end, going fast you sacrifice image quality.

Best, zdenek


On Sun, Jul 31, 2022 at 8:55 AM Emmanuel Levy <[log in to unmask]>
wrote:

> *****
> To join or leave the confocal microscopy listserv or to change your email
> address, go to:
> https://lists.umn.edu/cgi-bin/wa?SUBED1=confocalmicroscopy&A=1
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Dear Microscopists,
>
> We bought a spinning disk confocal microscope (Yokogawa W1 + Olympus X83).
> We have also ordered DIC components. However, the current setup requires
> moving a filter cube between DIC / fluorescent imaging. This is very
> problematic for us because it means 3 seconds lost per image set.
>
> Hence, I'm wondering if some of you are aware of a solution to this
> problem? e.g., would it be possible to make the polarization of the
> lasers + DIC light compatible and placing the analyzer in the filter wheel?
>
> If you have ideas or experience with such a question your feedback will be
> highly appreciated!
>
> Thank you,
> Best wishes,
>
> Emmanuel
>


-- 
-- 
Zdenek Svindrych, Ph.D.
Research Scientist - Microscopy Imaging Specialist
Department of Biochemistry and Cell Biology
Geisel School of Medicine at Dartmouth