LABEL-FREE, SUB-DIFFRACTION IDENTIFICATION OF BIOMOLECULES
Abstract
Research problem- In conventional bioimaging, fluorophores are routinely used to physically label proteins of interest. While fluorescence provides exceptional specificity, labeling may interfere with normal function. Moreover, fluorescence suffers from photobleaching, limiting time course studies and precluding a complete picture of biophysical processes. There is a need for new label-free bioimaging methods to visualize proteins below the diffraction limit in an unperturbed manner, preserving structure and functionality. Herein, the principal investigator proposes label-free sub-diffraction harnessing amplitude, phase and polarization information to resolve and identify individual proteins. The proposed method will meet the following criteria of label-free, sub-diffraction spatial resolution, large field of view, and high frame rate temporal resolution. Label-free sub-diffraction will allow for new opportunities to label-free nanoscopic observation of unperturbed proteins within their microenvironment. Technical approach- The objective is to enable sub-diffraction identification of biomolecules (e.g., proteins) in a label-free manner. By enabling sub-diffraction spatial resolution via polarization, the PI proposes to quantify amplitude and phase of scattered light from individual biomolecules (proteins). In a marker-free manner, the PI proposes to uniquely distinguish biomolecules. Finally, wide-field images of biomolecules will be pseudo-stained (i.e., electronically colored) via software for truly label-free sub-diffraction bioimaging. Anticipated outcome- The anticipated outcomes include- i. label-free, ii. pseudo-staining via software, iii. nonbleaching long term bioimaging, iv. sub-diffraction spatial resolution, v. large field of view, vi. high frame rate temporal resolution. DoD Impact- As a new biophysical tool, label-free sub-diffraction identification of proteins has the potential to enable pseudo-staining via software and eliminate the need for physical labeling, thereby preserving structure and functionality. Free from photobleaching, label-free subdiffraction over long observation time windows will enable long term visualization of biophysical processes.
Document Details
- Document Type
- DoD Grant Award
- Publication Date
- Mar 07, 2023
- Source ID
- FA95502210285
Entities
People
- Somin Eunice Lee
Organizations
- Air Force Office of Scientific Research
- Board of Regents of the University of Michigan
- United States Air Force