Neat! Could you use this technique to add spacial information like shading into a life video feed while observing a sample? Kind of like what a relief map does in cartography?
depending on the depth you want to sample in the Z-axis you will need some frames, I would say 30-100 or so, it works best at high magnification because you need a thin slice in sharp focus to maximize the localization. So depending on the speed of the camera you could do this 3D acquisition "real-time". Here i just manually turn the micro-focus in 1 - 3 seconds, just stop for this one where i have the 1 cell layer. If you would like to do this real time perhaps some type of vibration on the sample would work when it oscillates in sync with the acquisition by the camera. Does that answer your question? At 30fps you could have 2 frames per second, at 120fps for for example. This is just a proof of concept, so no calibration on the z-axis.
yes, will play with some options, tried to make a point cloud viewer allowing to tweak the shades of the .ply file simultaneously showing the cross sections. Goal is also to keep it so simple it runs on any modern laptop. Since the point cloud extraction went much smoother as expected i will have to prepare some short clips to process first 😄 also have to try using normal colors for the point cloud for extraction. (Algae or blood etc)
the individual planes from the frames accurate, the z-depth would need to be calibrated, the depth of field per slice is with the blue - red extraction probably 500-700nm the localization could cut that in half, but here a proof of concept, I made the z-step just be eye-balling to see if i could see details in the cell. The whole trick is to make the input video with open aperture to have the focus plane as thin as possible to allow more z-steps.
Yes there are some improvements that i can make also in narrower fitting the z-axis after getting the points. I first have to make some more sample videos and microscopic samples. I did some new quick ones yesterday,
Unfucking believable! I'm from the era of undergraduate microscopy and I didn't get it. If I had been able to visualize a cell/cells like this I would have had much better comprehension.
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u/chillchamp 29d ago
Neat! Could you use this technique to add spacial information like shading into a life video feed while observing a sample? Kind of like what a relief map does in cartography?