Custom Super-Resolution Microscope for the Structural Analysis of Nanostructures
Abstract
This DURIP award provides funds for the Principal Investigator to purchase the components to build a custom Super-Resolution Microscope, which will be used primarily for multiplexed, high-throughput characterization of two- and three-dimensional nanostructures built from nucleic acid building blocks. The components of the custom Super-Resolution Microscope include a super-resolution microscope system, an anti-vibration table, lasers, optical beam shutters, mirrors, and mirror mounts. Super-Resolution light microscopy is a complementary technique to Atomic Force Microscopy and Electron Microscopy for characterizing nanostructures in their native environment. It offers high resolution (less than 10 nm) and multiplex capability. The Principal Investigator~s laboratory has recently developed a ~super-resolution~ imaging technique, called DNA-Point Accumulation for Imaging in Nanaoscale Topography (DNA-PAINT), that uses stochastically occurring single-molecule fluorescence events over time, generated by the transient hybridization of fluorescently labeled ~imager oligonucleotides~ present in the imaging buffer to complementary ~docking strands~ present on the target structure to be imaged, to localize the position of each fluorophore molecule with high precision. These localizations are then used to construct a super-resolution image. The full programmability of the DNA-DNA interactions that drive DNA-PAINT offers the ability to perform multiplexed super-resolution imaging against 10 or more targets in the same sample (Exchange-PAINT). Additionally, the observed frequency of binding events over time can be harnessed to digitally count the number of docking strands present (qPAINT), and hence quantify the number of target moieties present in the assembled structures, which could be useful in cases where the same building block is used multiple times within a structure. The Principal Investigator has three Department of Defense (DoD)-funded research efforts (1 ONR, 1 ARO, and 1 AFOSR) that will benefit from the Super-Resolution Microscope built with the requested components. All of those research projects involve the design and assembly of large and complex 2D and 3D nanostructures built from nucleic acid building blocks, and the Super-Resolution Microscope will aid in the characterization of the structures produced. Additionally, the custom Super-Resolution Microscope will benefit two other ongoing, non-DoD funded, collaborative research efforts in the Principal Investigator~s laboratory. One of those projects addresses mapping the three-dimensional structure of the human genome using DNA-PAINT, and the other involves the development of a multiplexed, quantitative method for in situ proteomics. Specifically, Exchange-PAINT and qPAINT are being combined with antibody staining for multiplexed imaging of proteins in neuronal synapses. As a leader in the field of super-resolution imaging technologies, the Principal Investigator has many opportunities to work collaboratively with other laboratories on projects focused on nucleic acid nanotechnology or cellular imaging. However, access to a Super-Resolution Microscope is presently limiting those collaborations, and acquisition of the components to build a custom Super-Resolution Microscope would alleviate that problem. Beyond its value to the specific research efforts in which it will be used, the custom Super-Resolution Microscope will also play an important role in the science and technology education of the undergraduate students, graduate students, and postdoctoral fellows performing the research activities with the equipment. The Principal Investigator~s laboratory typically contains between twenty and thirty members, many of whom participate in imaging-related research.
Document Details
- Document Type
- DoD Grant Award
- Publication Date
- Sep 30, 2016
- Source ID
- N000141612563
Entities
People
- Peng Yin
Organizations
- Office of Naval Research
- President and Fellows of Harvard College
- United States Navy