Counting and estimating the cells coverage area

[Dakhil]

I am suffering from counting and estimating the cells coverage area in my bright field images. I am trying with imagej but I have no clue it is the right way to count the cells and roughly estimate the coverage area of these cells. Firstly, I am doing trial and error auto local threshold with different parameters until get something better then using the Analyze particles but it still unbelievable data. Could you please give me a piece of advise to overcome this big issue. I uploaded a sample of my images.

12.jpg

[Paul van Schayck]

Dear Dakhil,

Even from the image I have trouble seeing what are cells and what are
not. Is it possible to improve the contrast during acquisition?

Kind regards,

Paul

On Wed, Jun 18, 2014 at 1:57 PM, Dakhil
<[hidden email]> wrote:

> I am suffering from counting and estimating the cells coverage area in my
> bright field images. I am trying with imagej but I have no clue it is the
> right way to count the cells and roughly estimate the coverage area of these
> cells. Firstly, I am doing trial and error auto local threshold with
> different parameters until get something better then using the Analyze
> particles but it still unbelievable data. Could you please give me a piece
> of advise to overcome this big issue. I uploaded a sample of my images.
>
> 12.jpg <http://imagej.1557.x6.nabble.com/file/n5008276/12.jpg>
>
>
>
> --
> View this message in context: http://imagej.1557.x6.nabble.com/Counting-and-estimating-the-cells-coverage-area-tp5008276.html
> Sent from the ImageJ mailing list archive at Nabble.com.
>
> --
> ImageJ mailing list: http://imagej.nih.gov/ij/list.html

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[Dakhil]

Dear Paul,

Thanks for your interest. Unfortunately, the objective and the camera has been placed underneath a glass plate with thickness of  11 mm and the bright light source through the objective tube. The cells are 3T6 fibroblasts and I am measuring the rheological properties under oscillatory rheometer in parallel-disk configuration. The cells density are so high, more  than 90% confluence and I have to calculate the cell coverage area and number of cells per each image have been taken.

Regards.
Haider