Development of a Combined Atomic Force Microscopy and Side-View Imaging System for Mechanotransduction Research Public Deposited

Downloadable Content

Download PDF
Last Modified
  • March 20, 2019
Creator
  • Beicker, Kellie
    • Affiliation: College of Arts and Sciences, Department of Physics and Astronomy
Abstract
  • Key metrics for understanding cell response to mechanical stimuli include rearrangement of the cytoskeletal and nucleoskeletal structure, induced strains and biochemical distributions; however, structural information during applied stress is limited by our ability to image cells under load. In order to study the mechanics of single cells and subcellular components under load, I have developed a unique imaging system that combines an atomic force microscope (AFM) with both vertical light-sheet (VLS) illumination and a new imaging technique called PRISM – Pathway Rotated Imaging for Sideways Microscopy. The combined AFM and PRISM system facilitates the imaging of cell deformation in the direction of applied force with accompanying pico-Newton resolution force measurements. The additional inclusion of light-sheet microscopy improves the signal-to-noise ratio achieved by illumination of only a thin layer of the cell. This system is capable of pico-newton resolution force measurements with simultaneous side-view high frame rate imaging for single-molecule and single-cell force studies. Longer-term goals for this instrument are to investigate how external mechanical stimuli, specifically single-molecule interactions, alter gene expression, motility, and differentiation. The overall goal of my dissertation work is to design a tool useful for mechanobiology studies of single cells. This requires the design and implementation of PRISM and VLS systems that can be coupled to the standard Asylum AFM on inverted optical microscope. I also examine the strategy and implementation of experimental procedures and data analysis pipelines for single-cell and single-molecule force spectroscopy. These goals can be broken down as follows: • Performed single-molecule force spectroscopy experiments. • Performed single-cell force spectroscopy experiments. • Constructed and characterized the side-view microscopy system. • Applied combined AFM and side-vew microscopy system.
Date of publication
Keyword
DOI
Resource type
Rights statement
  • In Copyright
Advisor
  • Falvo, Michael
  • Superfine, Richard
  • Cheney, Richard
  • Mitran, Sorin
  • Cecil, Gerald
Degree
  • Doctor of Philosophy
Degree granting institution
  • University of North Carolina at Chapel Hill Graduate School
Graduation year
  • 2016
Language
Parents:

This work has no parents.

Items