# Open3D and confocal microscopy

Hi all,

I discovered open3D and I am trying to use it to render and analyze confocal microscope images without relying on commercial software. I have experience in 2D image analysis but I am new to 3D, I am looking to maybe find advice and share what I do.

I work with confocal microscopy, which is a technique that acquire an image in each focal plane of a sample. A typical datasets is composed by stack of images in which each stack represent a deeper position in a sample (the gif below shows a cell with vesicles acquired with a confocal microscope and rendered with 3D viewer in ImageJ).

For what I understand, in a case like this everything start by extracting a point cloud and then render the volumes from it.

To convert a stack of images to a point cloud, I iterate* through the stack of image and extract the coordinate and intensity** value for each pixel above a certain threshold.
My output look like:
[x, y, z, r, g, b]
…multiples rows later…
[xn, yn, zn, rn, gn, bn]

Usually the results are very dense point cloud mainly where are located structure of interest. What could be a strategy to render them and create volumes?

Cheers!
Kos

// I add these notes that might be helpful to somebody else //

1* To get the coordinates of pixel above a certain threshold in an image, I use
numpy.where(image > threshold)
that return two array, one for the x and y position. The depth (z-position) will depend on the stack number (each images is acquired at a further depth).

2* It’s trivial but took me some time to get to it, a grayscale image can be converted to RGB by putting the same grayscale intensity in each channel (R, G and B). Since o3D works with float divide the intensity by the byte resolution (e.g. divide the pixel intensity by 255 if you are in 8bit).

Hi,
You could check this opensource software : http://icy.bioimageanalysis.org/
vtk ((vtk.org)) is also used in 3d reconstruction from microscope data. You can have a look in their website.In vtk there are several visualization options, which you can try out…
Hope this helps you.

BR
Ravi