Compositions, systems, and methods for enhanced label-free and fluorescence-based detection of nanoparticles
AssigneeUnchained Labs
InventorsGeorge G. Daaboul, David S. Freedman, Amit Deliwala
Life of this patent
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Abstract
Presented herein are compositions, systems, and methods related to optical substrates that simultaneous (1) enhance a fluorescence signal emitted by a fluorophore and (2) enhance “contrast” signal that comprises scattered signal intensity over substrate reflectivity at a non-fluorescent wavelength. In certain embodiments, the optical substrate comprises a thin, transparent, dielectric layer. In alternative embodiments, the optical substrate comprises a stack of thin, transparent dielectric layers, for example, that is designed for both specific scattering enhancement and fluorescence enhancement.
Claims16
A method for detecting particles located substantially in a target plane above and in proximity to a top surface of a reflective substrate via simultaneous detection of fluorescence and single-particle scattering, the method comprising:
- (a) directing illumination light from an illumination source towards the top surface of the reflective substrate, thereby illuminating the particles along with the reflective substrate;
- (b) detecting, with one or more detectors, a portion of the illumination light that is scattered by the particles and reflected by the reflected substrate;
- (c) directing excitation light from an excitation source towards the top surface of the reflective substrate, wherein the excitation source is separate from the illumination source, and wherein the excitation light has a wavelength corresponding to an excitation band of a fluorescent species within or on a surface of the particles, thereby exciting the fluorescent species;
- (d) detecting, with one or more detectors, fluorescent light emitted from the fluorescent species as a result of excitation by the excitation light;
- and
- (e) processing, (i) data corresponding to the detected portion of the illumination light that is scattered by the particles and reflected by the reflective substrate and (ii) data corresponding to the detected fluorescent light and to locate at least a portion of the particles,
- wherein the reflective substrate comprises a multi-layered optical interference coating comprising a stack of three or more layers on top of a base layer, wherein a thickness of each of the one or more layers in the stack is less than or equal to about 200 nm.
Dependencies are inferred from the claim text.