Re: How can we Distinguish In/Out of focus cells in confocal microscope?

Posted by Joel Sheffield on May 25, 2014; 4:36pm
URL: http://imagej.273.s1.nabble.com/How-can-we-Distinguish-In-Out-of-focus-cells-in-confocal-microscope-tp5007911p5007912.html

Hi Mohamed,

You haven't specified the tissue, but I'll assume that both cells are
within the z range of the instrument.  If it were simply a focus issue, you
should be able to change the focal level (i.e. do a z scan) and see if you
get any changes.

Joel



Joel B. Sheffield, Ph.D
Department of Biology
Temple University
Philadelphia, PA 19122
Voice: 215 204 8839
e-mail: [hidden email]
URL:  http://astro.temple.edu/~jbs


On Sun, May 25, 2014 at 9:54 AM, Mohamed Tleis <[hidden email]> wrote:

> Dear ImageJ members,
>
> We have two channel images one of bright-field and one of Green
> Fluoresence Protein, acquired by a confocal microscope. Recently we are
> concerned about some cells having less fluoresent due to the fact that it
> is out of focus. In the bright-field channel it is a bit hard to
> distinguish between the in and out of focus cells. I wonder whether any of
> you came across a similar problem or knows of some literature about it.
>
> Thank you in Advance for your answers.
>
>
> Best Regards,
> M. Tleis
> Phd. Candidate
> LIACS, Leiden University
> The Netherlands.
>
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> ImageJ mailing list: http://imagej.nih.gov/ij/list.html
>

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