Correlative atomic force microscopy quantitative imaging-laser scanning confocal microscopy quantifies the impact of stressors on live cells in real-time

Scientific Reports
Supriya V BhatTanya E S Dahms

Abstract

There is an urgent need to assess the effect of anthropogenic chemicals on model cells prior to their release, helping to predict their potential impact on the environment and human health. Laser scanning confocal microscopy (LSCM) and atomic force microscopy (AFM) have each provided an abundance of information on cell physiology. In addition to determining surface architecture, AFM in quantitative imaging (QI) mode probes surface biochemistry and cellular mechanics using minimal applied force, while LSCM offers a window into the cell for imaging fluorescently tagged macromolecules. Correlative AFM-LSCM produces complimentary information on different cellular characteristics for a comprehensive picture of cellular behaviour. We present a correlative AFM-QI-LSCM assay for the simultaneous real-time imaging of living cells in situ, producing multiplexed data on cell morphology and mechanics, surface adhesion and ultrastructure, and real-time localization of multiple fluorescently tagged macromolecules. To demonstrate the broad applicability of this method for disparate cell types, we show altered surface properties, internal molecular arrangement and oxidative stress in model bacterial, fungal and human cells exposed to 2,4-dichl...Continue Reading

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Citations

Apr 8, 2020·Journal of Bacteriology·Albertus ViljoenYves F Dufrêne
Jul 17, 2020·Frontiers in Microbiology·Yves F Dufrêne, Albertus Viljoen
Aug 6, 2019·Frontiers in Bioengineering and Biotechnology·Danahe MohammedSylvain Gabriele
May 25, 2021·Frontiers in Cellular and Infection Microbiology·Supriya V BhatTanya E S Dahms

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Methods Mentioned

BETA
confocal microscopy
AFM
correlative

Software Mentioned

JPK
Graph Pad Prism
Photoshop

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