Super-resolution microscopy enables biologists to see the previously invisible by circumventing the classical diffraction limit, and holds promise to broadly transform biomedical research. However, current methods offer only limited resolution and multiplexing power, and tend to require either expensive instrumentation or specialized experimental conditions. They thus have not yet been widely adopted in standard biological labs. We propose a simple and robust super-resolution method based on the programmable autonomous blinking of a nucleic acid fluorescent probe. Unlike existing techniques that rely on the externally controlled stochastic blinking of special fluorophores, our technique utilizes programmable autonomous blinking of a nucleic acid (DNA/RNA) probe with prescribed brightness and blinking frequency. This unprecedented level of molecular control on the blinking behavior of the probe will enable an imaging technique that offers high multiplexing power and resolution. Unlike current super-resolution methods, our programmable blinking technique only requires standard instrumentation and is widely applicable and potentially genetically encodable. Development of this technique will bring the performance, usability, and applicability of super-resolution imaging to a new level, and help transform research practice in diverse biomedical fields.

Public Health Relevance

We propose to develop a new super-resolution imaging technique by engineering the blinking behavior of synthetic DNA/RNA imaging probes. Development of this technique will bring the performance, usability, and applicability of super-resolution imaging to a new level, and help transform research practice in diverse biomedical fields.

Agency
National Institute of Health (NIH)
Type
Research Project (R01)
Project #
5R01EB018659-02
Application #
8737257
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Conroy, Richard
Project Start
Project End
Budget Start
Budget End
Support Year
2
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Harvard Medical School
Department
Biology
Type
Schools of Medicine
DUNS #
City
Boston
State
MA
Country
United States
Zip Code
02115
Jungmann, Ralf; AvendaƱo, Maier S; Woehrstein, Johannes B et al. (2014) Multiplexed 3D cellular super-resolution imaging with DNA-PAINT and Exchange-PAINT. Nat Methods 11:313-8
Green, Alexander A; Silver, Pamela A; Collins, James J et al. (2014) Toehold switches: de-novo-designed regulators of gene expression. Cell 159:925-39
Iinuma, Ryosuke; Ke, Yonggang; Jungmann, Ralf et al. (2014) Polyhedra self-assembled from DNA tripods and characterized with 3D DNA-PAINT. Science 344:65-9