Near simultaneous laser scanning confocal and atomic force microscopy (Conpokal) on live cells

Joree N. Sandin, Surya P. Aryal, Thomas Wilkop, Christopher I. Richards, Martha E. Grady

Research output: Contribution to journalArticlepeer-review

7 Scopus citations

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.

Original languageEnglish
Article numbere61433
Pages (from-to)1-25
Number of pages25
JournalJournal of Visualized Experiments
Volume2020
Issue number162
DOIs
StatePublished - 2020

Bibliographical note

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ASJC Scopus subject areas

  • General Neuroscience
  • General Chemical Engineering
  • General Biochemistry, Genetics and Molecular Biology
  • General Immunology and Microbiology

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