(TKP2021-EGA-23) Támogató: Innovációs és Technológiai Minisztérium
Imaging of intracellular messengers, like calcium, is one of the most reliable methods
to follow real-time changes in several aspects of cellular activity, like receptor
activation. However, the analysis could be influenced and biased by several factors
like the location, shape, and size of the regions of interest (ROIs) and by the detection
and correction of the movement of the preparation. Programs which are provided by
the manufacturers are expensive and cannot be shared by collaborators. Many self-made
programs have been implemented lately which have in-built cell recognizer ROI identification
functions. These programs focus on the soma of the cells and neglect the processes,
because in full tissue preparation finding cells is still challenging. Subcellular
imaging experiments are still rare. To the best of our knowledge there is no program
which can automatically define ROIs for subcellular imaging experiments even in single
indicated cells with complex morphology. We developed and validated a program to address
this gap using simple and understandable mathematical methods for ROI determination
and simple statistics for movement correction. Validation experiments were conducted
on cochlear Deiters’ cells. Deiters’ cells have processed morphology which connects
two fluid compartments in the cochlea. Because of the function and the fine morphology
of the cell, it could be interesting to examine the subcellular Ca2+ handling mechanisms
of it. Test impulses were activated by ATP. With some limitations the program successfully
fulfilled its purpose. As a free, easily understandable, and open-source program,
we hope it will help to analyze and plan subcellular experiments.
