High content cellulomics by correlative microscopy

Abstract

The exponential increase in anthropogenic chemicals in our environment necessitates methods to assess impact prior to their release to predict potential environmental and health impacts. Atomic force microscopy (AFM) and laser scanning confocal microscopy (LSCM) each provide abundant information on cell behaviour. In addition to determining cell morphology and surface ultrastructure, AFM in quantitative imaging (QI) mode probes cellular mechanics and surface biochemistry with minimal applied force, and LSCM offers a window into the cell for imaging fluorescently tagged macromolecules. Data from correlative AFM-LSCM is thus complimentary, providing a comprehensive picture of cellular physiology. I will present a correlative AFM-QI-LSCM assay for the simultaneous real-time imaging of living cells in situ that generates real-time multiplexed data, including cell morphology and mechanics, surface adhesion and ultrastructure, along with the localization of intracellular macromolecules. The broad applicability of the assay is demonstrated using disparate cell types, showing altered surface properties, internal molecular arrangement and oxidative stress in model bacterial, fungal and human cells exposed to 2,4-dichlorophenoxyacetic acid. This correlative microscopy assay is broadly applicable to a variety of cell types and can be used to assess the impact of any multitude of contaminants, alone or in combination.