Posted by
Aryeh Weiss on
Jul 29, 2020; 1:39pm
URL: http://imagej.273.s1.nabble.com/Phase-contrast-image-segmentation-for-counting-fibroblasts-tp5023774p5023784.html
On 29/07/2020 15:40, Gabriel Landini wrote:
> On Wednesday, 29 July 2020 02:54:40 BST sfreema1 wrote:
>> I am told that the darkness in the center is a common observance when taking
>> phase contrast images in a 96-well plate.
> ~
> The image quality needs improving. I would start by learning how to adjust the
> phase rings and how to do background illumination correction.
> Without those two issues resolved you will struggle to get any data out.
>
> To answer the original question, here is a method we worked on to count cells
> in phase contrast images:
>
https://onlinelibrary.wiley.com/doi/pdf/10.1111/jmi.12726>
> Cheers
>
> Gabriel
>
> --
I will add that in 96 well plates, the meniscus displaces the image of
the phase ring. That is why the center of your image has better phase
contrast, and the edges are more like brightfield images. The Biovoxxel
Psuedo Flat Field Correction does a nice job on correcting the
background (and there are other methods that have been suggested a work
well). If you run Fiji you can add the Biovoxxel update site in the updater.
If you want to do phase contrast, I suggest that you restrict yourself
to the center of the well, and if possible also reduce the cell density
if possible.
--aryeh
--
Aryeh Weiss
Faculty of Engineering
Bar Ilan University
Ramat Gan 52900 Israel
Ph: 972-3-5317638
FAX: 972-3-7384051
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