Does anyone know a paper in which researchers created a real image or computational model of mitochondria dynamics in 3D in time. The papers discussing the problems of such analysis and image processing will be also interesting. It seems to me that the problem in such analysis is not the matter of speed of scanning and generally "hardware"but the post processing of the time lagged gathered images? Does anyone know of a paper which considers the problems of 3D time lapse imaging of complex structures moving in every dimension - like mitochondria? The most obvious to me are:
1. Lot of noise during fast Z-stacks capturing
2. Compensation for the time lag between images gathered in one Z stack in different focal planes - when we consider a one direction move of simple structure this seems to be easy but when mitochondria move fast in every dimension and are not homogeneous this seems to be really challenging.
I intuitively think that some kind of averaging of the changes in fluorescence in closest focal plains would allow such compensation of time lag to recreate the position of mitochondria in relation to the upper (imaged earlier) focal plains.
My previous lab imaged microtubule-bound EB1 comets in 3D cysts. They tend to move up and down in the z axis whilst moving in xy. I'm pretty sure they took rapid Z stacks (which requires a piezo motorized stage- like the ones from ASI) and created max intensity projections. There was a lot of optimisation to get the microscopy right, double GFP tagging EB1, a long pass eGFP filter etc... http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3350573/
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