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

Autor: Josef Lazar, Olga Rybakova, Jan Dohnálek, Alina Sakhi, Alexey Bondar, Ondřej Ticháček, Vendula Markova, Paul S. Miclea, Josef Melcr, Štěpán Timr, Petro Khoroshyy
Přispěvatelé: Molecular Dynamics
Jazyk: angličtina
Rok vydání: 2021
Předmět:
Computer science
QH301-705.5
Recombinant Fusion Proteins
Medicine (miscellaneous)
Molecular Dynamics Simulation
Linear dichroism
01 natural sciences
General Biochemistry
Genetics and Molecular Biology
Article
Fluorescence imaging
03 medical and health sciences
Protein structure
Single-cell analysis
GTP-Binding Proteins
Software Design
0103 physical sciences
Microscopy
Image Processing
Computer-Assisted
Humans
Biology (General)
Lipid bilayer
030304 developmental biology
Fluorescent Dyes
0303 health sciences
010304 chemical physics
Polarization Microscopy
Living systems
Luminescent Proteins
HEK293 Cells
Microscopy
Fluorescence
Microscopy
Polarization
Single-Cell Analysis
General Agricultural and Biological Sciences
Biological imaging
Biological system
Biological fluorescence
Signal Transduction
Zdroj: Communications biology, 4(1):189. Nature Publishing Group
Communications Biology, Vol 4, Iss 1, Pp 1-12 (2021)
Communications Biology
ISSN: 2399-3642
DOI: 10.1038/s42003-021-01694-1
Popis: 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
Databáze: OpenAIRE
Externí odkaz:
Nepřihlášeným uživatelům se plný text nezobrazuje