Re: widefield getting better images than spinning disk

Posted by RJ3 on
URL: http://confocal-microscopy-list.275.s1.nabble.com/widefield-getting-better-images-than-spinning-disk-tp7585177p7585192.html

*****
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.
*****

Thanks Sam,

http://imgur.com/a/KQysd​

Let me know if the above album works for you. I uploaded two images: one in
widefield and one in spinning disk confocal. All things else are equal. For
this example I used a fixed sample with 80 ms exposure. The image is at
10x, 0.45 NA with confocal pinhole of 40 um. Is this amount of image
degradation to be expected? I also tried turning up the light source
intensity significantly, with not much difference.

Rafael

On Fri, May 20, 2016 at 12:57 PM, Sam Lord <[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.
> *****
>
> On Fri, 20 May 2016 12:46:37 -0400, Rafael Jaimes III <[hidden email]
> >
> wrote:
> >1. Live neonatal rat cardiomyocyte-fibroblast layers stained with Fluo-4
> >for calcium. They are field stimulated and imaged >100 fps.
>
> At 10 ms exposure time, I doubt you're getting enough fluorescence hitting
> the
> camera to get a sufficient signal to noise. Wide field is always much
> brighter than
> confocal because both the excitation light and the emission intensity is
> reduced
> going through the pinholes.
>
> Why don't you send images of wide field vs confocal so we can see what you
> mean
> by "better."
>