SUMMARY OF INVENTION
Wide-field and confocal microscopes are limited by diffraction to approximately 250 nm lateral and 600 nm axial resolutions. Super-resolution microscopes achieve higher resolutions, however limitations still exist, including the gap that remains between spatial resolution and sizes of molecules and molecular complexes (particularly along the axial direction). Significantly, tradeoffs often must be balanced between spatial resolution, imaging speed, detection sensitivity, and photo-damage.
MSK investigators have demonstrated 3D super-resolution imaging by modulation interferometry. By utilizing two opposed lenses to create a structured illumination pattern and phase-sensitive detection, they have achieved localization of individual fluorophores with a level of precision smaller than 2nm. This is well below the 5-to-10 nanometer size of individual protein components of molecular complexes in cells.
This novel imaging system involves stable setup, and it is less susceptible to drifts and misalignments than existing technologies. It facilitates straightforward implementation for multicolor interferometry through the use of different excitation lasers, with no need for specialized detection optics tailored to each dye.
Highest resolution achieved in 3D super-resolution imaging
3D single-molecule localization with level of precision smaller than 2nm
Robust solution for interferometric imaging
Technology promises to be more affordable that current super-resolution microscopes
Broad potential applications for multicolor interferometry implementations
With a projected price of $200K-$300K, compared to the $500K-$1M cost for current super-resolution microscopes, this 3D imaging system could expand the market for high-resolution microscopes to include academic/nonprofit research laboratories as well as commercial end users.
AREAS OF APPLICATION
Imaging, research tool
STAGE OF DEVELOPMENT
Functional prototype developed
Pending application filed Dec. 2016
Alexandros Pertsinidis, PhD, Laboratory Head, Structural Biology Program, Memorial Sloan Kettering