Analysis of Mechanical Induction of Bioelectric Activity in Cells
Abstract
Major Goals: This project aims to use a combined imaging platform to examine how external stimuli are transduced by cellular structures to produce changes in internal electrical state via modulation of ion channel transmission. In previous work, the nanoscale sensitivity of quantitative phase imaging (QPI) has been used to reveal mechanical properties of cells. In this modality, an external shear stress is applied and sub-cellular changes in mass distribution are observed with QPI. A significant advantage of this assay is that the cells are not displaced during measurement. The platform was extended by incorporating FRET (Forster resonance energy transfer)- based sensors into the QPI platform. FRET technology reports the nanoscale separation of donor and acceptor molecules. In this approach, FRET is used to report local cell conditions such as the tension experienced by loadbearing proteins or the concentration of calcium ions. The combination of FRET-based sensors with QPI enables application of the approach to study dynamic loading of cells. Traditionally, such dynamic loading has required indirect measures and complex devices but with our platform, we can directly visualize cell stresses without mechanical displacement. To meet the goal of identifying how external stimuli are transduced by cellular structures to produce changes in internal electrical state via modulation of ion channel transmission, this project has adopted the following specific tasks: Task 1) Use measurements of FRET-based sensors and QPM to characterize signal transduction in cells under environmental challenges in ion concentration. In this task, we will develop methodology to analyze the mechanical signal transduction pathways which produce a change in cells internal state. Cells will be engineered to express FRET-based voltage and tension sensors.
Document Details
- Document Type
- Technical Report
- Publication Date
- Mar 11, 2023
- Accession Number
- AD1210763
Entities
People
- Adam Wax
- Brenton D. Hoffman
Organizations
- Duke University