(TKP2021-NVA-19) Támogató: Innovációs és Technológiai Minisztérium
Localization based superresolution technique provides the highest spatial resolution
in optical microscopy. The final image is formed by the precise localization of individual
fluorescent dyes, therefore the quantification of the collected data requires special
protocols, algorithms and validation processes. The effects of labelling density and
structured background on the final image quality were studied theoretically using
the TestSTORM simulator. It was shown that system parameters affect the morphology
of the final reconstructed image in different ways and the accuracy of the imaging
can be determined. Although theoretical studies help in the optimization procedure,
the quantification of experimental data raises additional issues, since the ground
truth data is unknown. Localization precision, linker length, sample drift and labelling
density are the major factors that make quantitative data analysis difficult. Two
examples (geometrical evaluation of sarcomere structures and counting the γH2AX molecules
in DNA damage induced repair foci) have been presented to demonstrate the efficiency
of quantitative evaluation experimentally.
