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

Sci Rep. 2020 May 18;10(1):8133. doi: 10.1038/s41598-020-65205-8.

Abstract

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.

Publication types

  • Evaluation Study
  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Actins / chemistry
  • Actins / metabolism
  • Animals
  • Biomechanical Phenomena
  • Fluorescence
  • HeLa Cells
  • Humans
  • Macrophages / chemistry*
  • Macrophages / cytology
  • Macrophages / metabolism
  • Mice
  • Microscopy, Atomic Force / methods*
  • Microscopy, Fluorescence / methods*
  • Phagocytosis
  • RAW 264.7 Cells

Substances

  • Actins