Quantitative linear dichroism imaging of molecular processes in living cells made simple by open software tools

0542218 - MBÚ 2022 RIV GB eng J - Článek v odborném periodiku
Bondar, Alexey - Rybakova, Olga - Melcr, J. - Dohnálek, J. - Khoroshyy, Petro - Ticháček, O. - Timr, S. - Miclea, Paul - Sakhi, A. - Marková, V. - Lazar, Josef
Quantitative linear dichroism imaging of molecular processes in living cells made simple by open software tools.
Communications Biology. Roč. 4, č. 1 (2021), č. článku 189. E-ISSN 2399-3642
Grant CEP: GA ČR(CZ) GJ17-14413Y
GRANT EU: European Commission(CZ) ATCZ14
Výzkumná infrastruktura: CESNET II - 90042; CERIT-SC - 90085
Institucionální podpora: RVO:61388971
Klíčová slova: linear dichroism * software tools * synthetic lipid vesicles * fluorescent proteins
Obor OECD: Biophysics
Impakt faktor: 6.548, rok: 2021
Způsob publikování: Open access
https://www.nature.com/articles/s42003-021-01694-1

Fluorescence-detected linear dichroism microscopy allows observing various molecular processes in living cells, as well as obtaining quantitative information on orientation of fluorescent molecules associated with cellular features. Such information can provide insights into protein structure, aid in development of genetically encoded probes, and allow determinations of lipid membrane properties. However, quantitating and interpreting linear dichroism in biological systems has been laborious and unreliable. Here we present a set of open source ImageJ-based software tools that allow fast and easy linear dichroism visualization and quantitation, as well as extraction of quantitative information on molecular orientations, even in living systems. The tools were tested on model synthetic lipid vesicles and applied to a variety of biological systems, including observations of conformational changes during G-protein signaling in living cells, using fluorescent proteins. Our results show that our tools and model systems are applicable to a wide range of molecules and polarization-resolved microscopy techniques, and represent a significant step towards making polarization microscopy a mainstream tool of biological imaging. Expanding on their previous work, Bondar et al present open source software tools to reliably quantify linear dichroism and determine molecular orientations. They demonstrate the utility of the tools by imaging synthetic lipid vesicles and as well as fluorescently labelled proteins in living cells
Trvalý link: http://hdl.handle.net/11104/0319688