Ultra-stable, photon-efficient cryogenic super-resolution fluorescence imaging for visualizing vitrified biological samples with molecular-scale resolution

NIH RePORTER · NIH · R21 · $215,825 · view on reporter.nih.gov ↗

Abstract

ABSTRACT Cryogenic electron microscopy (CryoEM) and cryo-electron tomography (CryoET) are powerful techniques for visualizing macromolecular assemblies at a near-native state and in their functional context inside cells; however the undiscriminating/non-specific contrast of such techniques often complicates identifying regions of interest and specific molecular targets in high-resolution cryoEM/cryoET datasets. Super-resolution (SR) fluorescence methods have exquisite molecular specificity, but SR techniques have only achieved limited spatial resolutions (~20-60 nm FWHM) when imaging vitrified biological samples at cryogenic temperatures. The proposed project will create new ultra-stable, photon efficient cryogenic SR methods for imaging vitrified biological specimens with true molecular-scale resolution (<2-3 nm FWHM), closing the gap towards the (sub-)nanometer resolution of cryoEM/cryoET. The increased spatial resolution will be leveraged for further development of powerful correlative light-electron microscopy (CLEM) approaches that will ultimately enable elucidating the structural basis for many critical macromolecular processes inside cells.

Key facts

NIH application ID
10510195
Project number
1R21GM147769-01
Recipient
SLOAN-KETTERING INST CAN RESEARCH
Principal Investigator
Alexandros Pertsinidis
Activity code
R21
Funding institute
NIH
Fiscal year
2022
Award amount
$215,825
Award type
1
Project period
2022-09-20 → 2024-08-31