Abstract
Techniques available for micro- and nano-scale mechanical characterization have exploded in the last few decades. From further development of the scanning and transmission electron microscope, to the invention of atomic force microscopy, and advances in fluorescent imaging, there have been substantial gains in technologies that enable the study of small materials. Conpokal is a portmanteau that combines confocal microscopy with atomic force microscopy (AFM), where a probe "pokes" the surface. Although each technique is extremely effective for the qualitative and/or quantitative image collection on their own, Conpokal provides the capability to test with blended fluorescence imaging and mechanical characterization. Designed for near simultaneous confocal imaging and atomic force probing, Conpokal facilitates experimentation on live microbiological samples. The added insight from paired instrumentation provides co-localization of measured mechanical properties (e.g., elastic modulus, adhesion, surface roughness) by AFM with subcellular components or activity observable through confocal microscopy. This work provides a step by step protocol for the operation of laser scanning confocal and atomic force microscopy, simultaneously, to achieve same cell, same region, confocal imaging, and mechanical characterization.
Document Type
Article
Publication Date
8-11-2020
Digital Object Identifier (DOI)
https://doi.org/10.3791/61433
Funding Information
We acknowledge NIH COBRE funding under number P20GM130456.
Repository Citation
Sandin, Joree N.; Aryal, Surya P.; Wilkop, Thomas E.; Richards, Christopher I.; and Grady, Martha E., "Near Simultaneous Laser Scanning Confocal and Atomic Force Microscopy (Conpokal) on Live Cells" (2020). Physiology Faculty Publications. 163.
https://uknowledge.uky.edu/physiology_facpub/163
Included in
Biomedical Engineering and Bioengineering Commons, Chemistry Commons, Mechanical Engineering Commons, Physiology Commons
Notes/Citation Information
Published in Journal of Visualized Experiments, issue 162, e61433.
Copyright © 2020 JoVE
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