- Replaced Matlab’s iteration to a custom one for increased reliability when calculating scalar or vectorial PSFs.
- Modification to the focal position determination when calculating scalar or vectorial PSFs. Results only slightly different focal position than the previous algorithm, in normal cases (~22 nm difference with the default settings).
- Fixed a bug when calculating scalar or vectorial PSFs (not sure whether a new Matlab version caused it or it was originally in the code). The PSFs shapes and peaks were only marginally affected.
- Made a workaround for a bug causing higher blinking event brightness when simulating with very low “Sample depth” value.
- Added “Synaptosomes” option to the random “Vesicles” pattern.
- Parameters files exported with previous TestSTORM versions (2.0.x) and containing “Rods”, “Vesicles” or “Octagons” patterns can not be imported any longer starting this version.
- Added a tool with which the generated PSF can be depicted.
Other clean-ups and modification in the code which should not affect the behaviour of the program.
Tamás Gajdos won a grant at the National Talent Program (Nemzeti Tehetség Program). He will cover the MBFT XVII. Congress registration fee, a german language course and an IT upgrade by this grant.
TestSTORM: Versatile simulator software for multimodal super-resolution localization fluorescence microscopy
Optimization of sample, imaging and data processing parameters is an essential task in localization based super-resolution microscopy, where the final image quality strongly depends on the imaging of single isolated fluorescent molecules. A computational solution that uses a simulator software for the generation of test data stacks was proposed, developed and tested. The implemented advanced physical models such as scalar and vector based point spread functions, polarization sensitive detection, drift, spectral crosstalk, structured background etc., made the simulation results more realistic and helped us interpret the final super-resolved images and distinguish between real structures and imaging artefacts.
Novák et al, Scientific Reports 7, Article number: 951 (2017) doi:10.1038/s41598-017-01122-7
Miklós Erdélyi, József Sinkó, Tamás Gajdos and Tibor Novák – “Enhanced simulator software for image validation and interpretation for multimodal localization super-resolution fluorescence microscopy “, Proc. SPIE 10071, Single Molecule Spectroscopy and Superresolution Imaging X, 100710X (February 21, 2017); doi:10.1117/12.2250116;
A practical method has been presented for polarization sensitive localization based super-resolution microscopy using a birefringent dual wedge. The measurement of the polarization degree at single molecule level can reveal the chemical and physical properties of the local environment of the fluorescent dye molecule and can hence provide information about the sub-diffraction sized structure of biological samples. Polarization sensitive STORM imaging of the F-actins proved correlation between the orientation of fluorescent dipoles and the axis of the fibril.