Advancing Visualization and Quantification of Subcellular and Biomolecular Mechanics through Mechanochemical Protocols - Project Summary
Exploring cellular mechanics represents a groundbreaking frontier to unravel the complexities of life and
disease. Far from being static entities, cells are dynamic, active materials that generate and sustain mechanical
forces, serving as critical indicators for various pathologies. Despite remarkable strides in cell mechanics, we
stand in a new era that beckons us to surmount many challenges that have long stymied progress. These
challenges span from technological obstacles to limited versatile methodologies, each representing a unique
puzzle and challenge in biomechanics.
Our MIRA program embarks on an unprecedented journey to address biomechanics challenges by
leveraging an interdisciplinary approach that melds the strengths of chemistry and mechanical engineering. The
PI's expertise in mechanochemistry, mechanical sensing, and fluorescent microscopy has positioned him as a
suitable researcher to use mechanochemical protocols to visualize biomechanical mapping into subcellular and
molecular levels. We will address the following challenges: First, the cell mechanics field lacks a versatile
technique for quantifying multiscale cell mechanics. Existing methods like Atomic Force Microscopy (AFM) and
Fluorescence Resonance Energy Transfer (FRET) offer partial solutions but come with their own sets of
limitations, such as intricate setups, specialized expertise, and low throughput. Second, high-precision, real-time
force measurements at the nanoscale are missing. While AFM provides nanoscale resolution, its limitations for
in vivo applications and the viscoelastic nature of cells add layers of complexity that further complicate data
interpretation. Third, the obstacle of accurately measuring in vivo dynamic biomechanical regulations is a
significant hurdle in cell mechanics. Existing techniques, although precise, are generally unsuitable for in vivo
applications due to their invasive nature and complex setups. The dynamic milieu of living systems further
complicates the issue, demanding rapid and continuous measurements. These challenges are not mere
obstacles; they are opportunities, beckoning us to innovate, explore, and revolutionize biomechanics. As we
embark on this exhilarating journey, we are not just solving puzzles but pushing the boundaries of what is
possible, opening new avenues for research and clinical applications.
This MIRA program will establish a pioneering analytical platform for biomechanics, from building a
comprehensive library of Molecular Force Gauges (MFGs), integrating these force gauges into various biological
targets for precise biomechanical measurements, to developing a state-of-the-art super-resolution
biomechanical microscopy BM-STORM to visualize and quantify cellular and subcellular biomechanics with
unprecedented detail. Our research agenda will be based on a three-fold approach. First, we will pioneer a
comprehensive library of Molecular Force Gauges (MFGs) designed to offer unprecedented insights into cellular
and subcellular mechanics. These MFGs are ingeniously engineered to undergo optical or luminescent
transformations in response to mechanical forces, serving as real-time, nanoscale force sensors. Our approach
addresses the limitations of existing technologies like flipper probes and FRET, offering a more direct and
versatile method for force quantification. Second, we will utilize these MFGs to sense both compressive and
tensile forces across various biological targets. This dual-mode operation enriches our understanding of
membrane mechanics and offers a robust and reliable metric for tension measurements. Third, we will develop
an innovative super-resolution biomechanical microscopy, BM-STORM, to visualize and quantify cellular and
subcellular biomechanics with unprecedented detail. This system leverages mechanical force to control the
switching between bright and dark states of MFGs, offering a biomechanical counterpart to Stochastic Optical
Reconstruction Microscopy (STORM). Our multidimensional approach will address the most pressing challenges
in biomechanics, from multiscale force quantification to real-time, in vivo imaging. Through these innovations,
we will elevate the field of biomechanics to an unprecedented level.
Our approach is inherently flexible, allowing for the seamless incorporation of other possible MFGs and the
adoption of other new super-resolution microscopy techniques. Our long-term vision, spanning 5 to 20 years, is
to evolve this biomechanical measurement platform into a robust, reliable, and versatile set of protocols. This
will pave the way for groundbreaking advancements, including in vivo biomechanical testing, three-dimensional
biomechanical mapping, and deep-tissue biomechanical monitoring.
