[Yeast] halo effect in fluorescent channel

Dear all,

I am having some problem with image analysis and I wander if I can find a set of modules in CP which might resolve my issues.

I am doing a timelapse images of Yeast colony grow using epifluorescent microscope. We induce the fluorescence which grows and decreases during the acquisition which takes around 2 days in microfluidic chamber. We use same exposure for every image. We compute the fluorescent for each cell during this period. However with the time when the signal get stronger we observe a kind of “halo effect around whole colony”. Therefore we think that we overestimate the signal in the cells in the center, since they are getting the most fake signal from “halo gradient”. Please check the image I attached.

Is there anyway to remove this halo effect with a combination of CP modules? Is it known phenomena with which one can easily deal in any other way? Is it the wrong setting of microscope or something which is standard and we can not avoid?

Thanks a lot for any comment or suggestion.

Szymon STOMA
INRIA Paris-Rocquencourt
Domaine de Voluceau BP 105
78153 Le Chesnay Cedex
Tel: +33 659 777 221
Web: stoma.name/

I am not sure that the attachment was not corrupted last time. I converted it to png.

Hi Szymon,

Would you mind posting a comparable picture in which the gradient is not present (I presume from earlier in the sequence?) I can take a look and assess the situation but I would need an example of both extremes as I can compare and contrast…