Combined Atomic Force Microscope and Volumetric Light Sheet System for Correlative Force and Fluorescence Mechanobiology Studies.

Journal Article (Journal Article)

The central goals of mechanobiology are to understand how cells generate force and how they respond to environmental mechanical stimuli. A full picture of these processes requires high-resolution, volumetric imaging with time-correlated force measurements. Here we present an instrument that combines an open-top, single-objective light sheet fluorescence microscope with an atomic force microscope (AFM), providing simultaneous volumetric imaging with high spatiotemporal resolution and high dynamic range force capability (10 pN - 100 nN). With this system we have captured lysosome trafficking, vimentin nuclear caging, and actin dynamics on the order of one second per single-cell volume. To showcase the unique advantages of combining Line Bessel light sheet imaging with AFM, we measured the forces exerted by a macrophage during FcɣR-mediated phagocytosis while performing both sequential two-color, fixed plane and volumetric imaging of F-actin. This unique instrument allows for a myriad of novel studies investigating the coupling of cellular dynamics and mechanical forces.

Full Text

Duke Authors

Cited Authors

  • Nelsen, E; Hobson, CM; Kern, ME; Hsiao, JP; O'Brien Iii, ET; Watanabe, T; Condon, BM; Boyce, M; Grinstein, S; Hahn, KM; Falvo, MR; Superfine, R

Published Date

  • May 18, 2020

Published In

Volume / Issue

  • 10 / 1

Start / End Page

  • 8133 -

PubMed ID

  • 32424215

Pubmed Central ID

  • PMC7234992

Electronic International Standard Serial Number (EISSN)

  • 2045-2322

Digital Object Identifier (DOI)

  • 10.1038/s41598-020-65205-8

Language

  • eng

Conference Location

  • England