Quantifying Nuclei to Golgi angle

Hi all,

I am trying to quantify cell orientation by measuring the nuclei to golgi angle in endothelial cells. At the moment, I am doing it manually and so can only do about a 750-1000 cells per sample. I have images containing ~10000 cells and would like to automate this process to expand my analysis.

I have included a sample image where golgi is green and nuclei are blue.

Any suggestions on how to achieve this? I have a basic understanding of ImageJ macros and segmentation.

A1.tif (2.9 MB)

What is the color of Golgi? Do you have the individual channel image for them?

The golgi is in green. I have split the channel here.

A1 Golgi.tif (976.8 KB)
A1 Nuclei.tif (976.8 KB)

Hello all,

I have to ask, how are you defining the angle? An angle is usually between 3 points, so are you asking for the angle between nucleus center to golgi center(s) with the respect to the horizon? Could you please share an illustration on how you are computing this angle manually, to get a better idea of the procedure?

Best

Hi Olivier,

Thank you for your interest.

These cells are grown in chambers through which we can flow media. The shear stress from the flow polarizes the cells and that is reflected on the positioning of the golgi relative to the nuclei. In the images, the axis of flow is vertical. So during the initial measurement any axis can be used and then I can normalize it to the direction of the flow.

I hope this clarifies everything for you.

Thanks

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