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.
|Number of pages||25|
|Journal||Journal of Visualized Experiments|
|State||Published - 2020|
Bibliographical noteFunding Information:
We acknowledge NIH COBRE funding under number P20GM130456. We thank the UK Light Microscopy Core, which is supported by the Vice President for Research, for assistance in this work. Strains of S. mutans and E. faecalis were provided by Dr. Natalia Korotkova.
© 2020 JoVE Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License.
ASJC Scopus subject areas
- Neuroscience (all)
- Chemical Engineering (all)
- Biochemistry, Genetics and Molecular Biology (all)
- Immunology and Microbiology (all)