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

Posted by Joel Sheffield on May 25, 2014; 6:16pm
URL: http://imagej.273.s1.nabble.com/How-can-we-Distinguish-In-Out-of-focus-cells-in-confocal-microscope-tp5007911p5007915.html

Hi Mohamed,

It would help if you could post an example image so that we could see what
you are dealing with.

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 1:00 PM, Mohamed Tleis <[hidden email]> wrote:

> Dear Joel,
>
> Thank you for your reply, My Cells are simple yeast cells. I do Image
> Analysis for some images acquired by Biologists. they try to  change the
> focal level to the most optimal one then they acquire their images.
> However, In the analysis part we need to be aware of out of focus cells,
> because they can wrongly affect the final measurement results. So my
> question is: is there any means to deal with such issues?
>
>
> Best Regards,
>
> M. Tleis
> Phd. Candidate
> LIACS, Leiden University
> The Netherlands.
>
>
>
>
>
> On 05/25/2014 06:36 PM, JOEL B. SHEFFIELD wrote:
>
>> 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.
>>>
>>> --
>>> ImageJ mailing list: http://imagej.nih.gov/ij/list.html
>>>
>>>  --
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>>
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