Dear M. Tleis,
As far as I know the image of the fluorescence
signal is confocal and always is on focus while
bright field image is not confocal (I mean is not
a thin optical section). If there are a lot of
cells and you are not sure in which cell is the
fluoresence signal is, what you can do (or ask
for) is a stack (let´s say three or four images
in Z) instead of a single image of the area.
Maybe they can give you a better sight. I hope this helps you.
B. Alarcón, phD
Instituto de Biomedicina-CSIC
Valencia, Spain
> 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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