Molecular and cellular level characterization of cytoskeletal mechanics using a quartz crystal microbalance

Abstract

A quartz crystal microbalance (QCM) is an instrument that has the ability to measure nanogram‐level changes in mass on a quartz sensor and is traditionally used to probe surface interactions and assembly kinetics of synthetic systems. The addition of dissipation monitoring (QCM‐D) facilitates the study of viscoelastic systems, such as those relevant to molecular and cellular mechanics. Due to real‐time recording of frequency and dissipation changes and single protein‐level precision, the QCM‐D is effective in interrogating the viscoelastic properties of cell surfaces and in vitro cellular components. However, few studies focus on the application of this instrument to cytoskeletal systems, whose dynamic parts create interesting emergent mechanics as ensembles that drive essential tasks, such as division and motility. Here, we review the ability of the QCM‐D to characterize key kinetic and mechanical features of the cytoskeleton through in vitro reconstitution and cellular assays and outline how QCM‐D studies can yield insightful mechanical data alone and in tandem with other biophysical characterization techniques.

Deciphering Mechanochemical Influences of Emergent Actomyosin Crosstalk using QCM-D

“…A quartz crystal microbalance with dissipation monitoring (QCM-D) is an instrument that uses a piezoelectric sensor to detect ng-level changes in mass, as well as changes in viscoelasticity in systems attached to the sensor surface. [32][33][34] While QCM-D has traditionally been used to study properties of coatings and polymers, it has started to gain traction in the biophysics field to study changes in mechanical properties of biological materials. 32,35 Applications for QCM-D in studying the cytoskeleton have been reviewed by Kerivan et al 32 Most cytoskeletal applications involve studying how cells deform in response to a stimulus or drug.…”

“…[32][33][34] While QCM-D has traditionally been used to study properties of coatings and polymers, it has started to gain traction in the biophysics field to study changes in mechanical properties of biological materials. 32,35 Applications for QCM-D in studying the cytoskeleton have been reviewed by Kerivan et al 32 Most cytoskeletal applications involve studying how cells deform in response to a stimulus or drug. There have been in vitro cytoskeletal studies that focus more on binding mechanisms of motors and filaments, as well as gaining a better understanding of how classic in vitro motility assays assemble and behave on a non-physiological surface, like a coverslip.…”

“…Until improved models are developed for quantitatively interpreting QCM-D frequency and dissipation change measurements for cytoskeletal ensembles, it would be beneficial to couple the qualitative information regarding changes in assay conditions and system viscoelasticity with quantitative force measurement techniques, like AFM or optical tweezers, or in conjunction with fluorescence microscopy. 32 For example, Yang et al characterized cell-level mechanical behavior of epithelial cells in response to epidermal growth factor (EGF) using a combination of fluorescence imaging to capture the response of actin filaments, AFM to probe the cell's mechanical response, and QCM-D to correlate energy with the measured AFM response to EGF stimulation. 50 Their approach demonstrates the advantage of using complementary techniques to formulate a more complete biophysical profile of their system of interest and should be applied more readily to in vitro studies.…”

“…32–34 While QCM-D has traditionally been used to study properties of coatings and polymers, it has started to gain traction in the biophysics field to study changes in mechanical properties of biological materials. 32,35 Applications for QCM-D in studying the cytoskeleton have been reviewed by Kerivan et al . 32 Most cytoskeletal applications involve studying how cells deform in response to a stimulus or drug.…”

“…32,35 Applications for QCM-D in studying the cytoskeleton have been reviewed by Kerivan et al . 32 Most cytoskeletal applications involve studying how cells deform in response to a stimulus or drug. There have been in vitro cytoskeletal studies that focus more on binding mechanisms of motors and filaments, as well as gaining a better understanding of how classic in vitro motility assays assemble and behave on a non-physiological surface, like a coverslip.…”