Super-resolution microscopy using tunable fluorescent nanodiamonds

This technology is an imaging technique called deterministic emitter switch microscopy (DESM) that employs tunable fluorescent nanodiamonds to achieve nanometer-scale fluorescence imaging.

Unmet Need: Super-resolution microscopy for rapid, wide-field imaging

Current super-resolution microscopy methods, such as photoactivated localization microscopy (PALM) and stochastic optical reconstruction microscopy (STORM), use sequential activation of photoswitchable fluorophores to produce high-resolution images well below the diffraction limits of traditional fluorescence microscopy. However, these methods are limited by the need for precise control over the density of fluorophores, and suffer from trade-offs between photostability and imaging rate. As such, there is a need for a microscopy method that enables rapid, super-resolution imaging of photostable and controllable fluorophores.

The Technology: Super-resolution nanometer-scale microscopy using externally-controlled fluorescent nanodiamonds

This technology uses externally-controlled fluorescent nanodiamonds to enable rapid, super-resolution microscopy. This technology works by selectively exciting nitrogen vacancy centers (NVCs) within fluorescent nanodiamonds using specific light or microwave field frequencies. Images are then captured by a CCD camera and processed to resolve the signal from individual fluorescent nanodiamonds. Using this process, this technology can distinguish up to an estimated 10,000 NVCs within a focal spot to produce a super-resolution image. Combined with the ability of NVCs to sense magnetic and electric fields, this technology enables rapid, super-resolution imaging for tracking and sensing applications in the life and physical sciences.

This technology has been used to resolve fluorescent images down to 12 nm across a 35 × 35 micron area.

Applications:

• Nanometer-range super-resolution fluorescence imaging
• Multicolor particle tracking and imaging
• High-resolution electric or magnetic field sensor
• High-resolution biological imaging

Advantages:

• Nanodiamonds are biocompatible, non-bleaching, and bright
• Can image up to 10,0004 distinct points within a focal spot
• Compatible with confocal microscopy
• May also be used to image NVCs in bulk diamond or other emitters with externally-controllable fluorescence

Lead Inventor:

Dirk Englund, Ph.D.

Patent Information:

Patent Pending (US 20160161429)

Related Publications:

• Karaveli S, Gaathon O, Wolcott A, Sakakibara R, Shemesh OA, Peterka DS, Boyden ES, Owen JS, Yuste R, Englund D. “Modulation of nitrogen vacancy charge state and fluorescence in nanodiamonds using electrochemical potential” Proc Natl Acad Sci USA. 2016 Apr 12;113(15):3938-43.

• Trusheim ME, Li L, Laraoui A, Chen EH, Bakhru H, Schröder T, Gaathon O, Meriles CA, Englund D. “Scalable fabrication of high purity diamond nanocrystals with long-spin-coherence nitrogen vacancy centers” Nano Lett. 2014 Jan 8;14(1):32-6.

• Chen EH, Gaathon O, Trusheim ME, Englund D. “Wide-Field Multispectral Super-Resolution Imaging Using Spin-Dependent Fluorescence in Nanodiamonds” Nano Lett. 2013 May 8;13(5):2073-7.

Tech Ventures Reference:

• IR CU12035, IR CU12035-a

• Licensing Contact: Greg Maskel