A Method for High‐Throughput Measurements of Viscosity in Sub‐micrometer‐Sized Membrane Systems

To unravel the underlying principles of membrane adaptation in small systems like bacterial cells, robust approaches to characterize membrane fluidity are needed. Currently available relevant methods require advanced instrumentation and are not suitable for high‐throughput settings needed to elucida...

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Bibliographic Details
Published in:Chembiochem : a European journal of chemical biology 2020-03, Vol.21 (6), p.836-844
Main Authors: Chwastek, Grzegorz, Petrov, Eugene P., Sáenz, James Peter
Format: Article
Language:English
Subjects:
Adaptation
bacteria
Emission measurements
Fluidity
Fluorescence
Fluorescent dyes
Fluorescent indicators
high-throughput screening
Instrumentation
Lipid membranes
Lipids
Membrane fluidity
Membranes
Phase transitions
Robustness
spectroscopy
Viscosity
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Summary:To unravel the underlying principles of membrane adaptation in small systems like bacterial cells, robust approaches to characterize membrane fluidity are needed. Currently available relevant methods require advanced instrumentation and are not suitable for high‐throughput settings needed to elucidate the biochemical pathways involved in adaptation. We developed a fast, robust, and financially accessible quantitative method to measure the microviscosity of lipid membranes in bulk suspension using a commercially available plate reader. Our approach, which is suitable for high‐throughput screening, is based on the simultaneous measurements of absorbance and fluorescence emission of a viscosity‐sensitive fluorescent dye, 9‐(2,2‐dicyanovinyl)julolidine (DCVJ), incorporated into a lipid membrane. We validated our method using artificial membranes with various lipid compositions over a range of temperatures and observed values that were in good agreement with previously published results. Using our approach, we were able to detect a lipid phase transition in the ruminant pathogen Mycoplasma mycoides. Membrane adaptation: We developed a robust and financially accessible quantitative method to measure the microviscosity of lipid membranes that is suitable for high‐throughput screening. The approach is based on the simultaneous measurements of absorbance and fluorescence emission of a fluorescent dye and allows monitoring of the temperature dependence of viscosity in bacterial membranes.
ISSN:1439-4227
1439-7633
DOI:10.1002/cbic.201900510