Technology Development Plan (TDP)
Our solution to the technical limitations of SMFS and TFM is the molecular tension probe (MTP). In 2011, the lab of the PI (Salaita) pioneered the development of molecular tension fluorescence microscopy (MTFM) where a probe comprised of a fluorophore–quencher pair separated by a molecular “spring” is immobilized onto a surface. Despite the impact and rapid growth of MTP technologies, there remain some major gaps in our capabilities. Each TDP is designed to address these issues and uses a range of prototypes that have progressed beyond the proof-of-concept validation stage. The goal of the CPMM is to transform the field of mechanobiology, and we will do so by addressing the technological gaps.
TDP#1 Probes for High Resolution Mechanobiology
The goal of this TDP is to transform the research accessibility of DNA molecular tension probes (MTP) and advance its capabilities, allowing orders of magnitude improvement in probe stability, spatial resolution, and orientational resolution. The work will help broaden the accessibility of DNA MTP to the research community and enable new biomedical applications, aligning with the long-term goals of the BTDD.
Adapted from Salaita, Mattheyses & Ke Nature Methods 2020.
TDP#2 Probes for Mechanical Identification
Flow cytometry is routinely used to measure single-cell gene expression by staining cells with fluorescent antibodies and nucleic acids. Here we present Tension-activated Cell Tagging (TaCT) to fluorescently label cells based on the magnitude of molecular force transmitted through cell adhesion receptors. As a proof-of-concept, we analyzed fibroblasts and mouse platelets after TaCT using conventional flow cytometry.
Adapted from Salaita et al. 2023 Nat. Methods
Harnessing cellular force measurements for applications such as diagnosis of disease and drug screening requires ultrasensitive and highly specific tension probe signal. Under TDP#3, we will leverage the programmability of DNA to generate tension probes with enhanced sensitivity and amplified response to mechanical input. The common theme in this TDP is not a specific driving biomedical question, rather the uniting theme is overcoming the common challenge of detecting highly transient biophysical events that are sparse and distributed across the cell surface.
