Evaluation of immunolabeling at electron microscopic level – introduction of a new online software tool

0555835 - ÚMG 2022 RIV CZ eng O - Others
Filimonenko, Vlada - Pinkas, Dominik - Vlčák, Erik - Raabová, Helena - Janáček, J. - Hozák, Pavel
Evaluation of immunolabeling at electron microscopic level – introduction of a new online software tool.
2021
R&D Projects: GA MŠMT(CZ) LM2018129; GA MŠMT(CZ) EF16_013/0001775; GA TA ČR(CZ) TE01020118
Research Infrastructure: Czech-BioImaging II - 90129
Institutional support: RVO:68378050
Keywords : electron mcirosopy * immunolabeling * statistical evaluation * online software tool
OECD category: Cell biology

Evaluation of immunolabelling in electron microscopy still often lacks unbiased quantitative approach.We have developed an easy-to-use online tool for semi-automatic multi-stage analysis of immunolabeling on EM images spanning particle detection and classification, mathematical and statistical evaluation and visualization of results. This builds on algorithms from previous work of A. Philimonenko et al. [1], C. Schofer et al. [2], and L. Pastorek et al. [3]. The tool detects gold nanoparticles of defined size automatically, results of detection and identification can be manually reviewed and edited. Spatial relations can be analyzed in 2D and 3D microscopic data and along linear structures such as membranes and filaments. The tool uses pair correlation and pair cross-correlation functions for clustering and colocalization evaluation with results presented both numerically and graphically. Labelled structures are visualized via
mapping. Statistical significance of detected patterns is calculated and presented in a comprehensive way without the need for deep insight into statistical analysis. All results and respective settings can be exported. Projects can be shared with colleagues. The tool has been released at the end of the year 2020. It is available to the broad scientific community in open access mode by the IMG within the Czech-BioImaging research infrastructure. The tool emphasizes convenience and understandability of the interface. The platform is modular and can be expanded with more capabilities in the future.
Permanent Link: http://hdl.handle.net/11104/0330294