The proposed research focuses on development of super-resolution optical imaging methods that utilize reversible and transiently binding probes as the labeling reagents. Using of reversibly-binding probes has the potential of achieving significantly higher labeling density and enhanced spatial resolution. However, currently their application is limited by the lack of suitable labeling reagents towards important cellular targets. Here we propose to expand the repertoire of diffusive labeling tools by developing new labeling methods to facilitate broader usages of PAINT methodology in biological studies and new statistical algorithms to achieve the optimal spatial resolution from localization dataset collected with diffusive labeling. Specifically we will focus on (1) developing imaging probes for phosphotyrosine signaling proteins based on Src- homology 2 (SH2) domains, (2) developing a hybrid genetic-chemical labeling method based on protein-peptide interactions that allows imaging of cellular proteins, and (3) developing statistical models and numerical algorithms to achieve resolution enhancement when target molecules are labeled in multiple rounds with reversible probes.

Public Health Relevance

Technology allowing high resolution localization of cellular molecules is one of the key driving forces in new biological discoveries. Here we seek to develop new experimental reagents and statistical algorithms that facilitate optical imaging of cellular proteins and their post-translational modification at unprecedented spatial resolutions.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM123784-02
Application #
9404037
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Sammak, Paul J
Project Start
2017-01-01
Project End
2020-12-31
Budget Start
2018-01-01
Budget End
2018-12-31
Support Year
2
Fiscal Year
2018
Total Cost
Indirect Cost
Name
University of Connecticut
Department
Biochemistry
Type
Schools of Medicine
DUNS #
022254226
City
Farmington
State
CT
Country
United States
Zip Code
06030