Velocimetry-based identification of single proteins and other particles

Patent metadata

Field	Value
Publication number	US-11717828-B2
Application number	US-201716471413-A
Country	US
Kind code	B2
Filing date	Dec 19, 2017
Priority date	Dec 20, 2016
Publication date	Aug 8, 2023
Grant date	Aug 8, 2023

Abstract

Spatially distributed optical excitation and integrated waveguides are used for ultrasensitive particle detection based on individual electrokinetic velocities of particles. In some embodiments, chip-integrated systems are used to identify individual particles (e.g., individual molecules) based on their velocity as they move through an optically interrogated channel. Molecular species may be identified and quantified in a fully integrated setting, allowing for particle analysis including molecular analysis that can operate at low copy numbers down to the level of single-cell lysates. In some embodiments, the single-particle velocimetry-based identification and/or separation techniques are applied to various diagnostic assays, including nucleic acids, metabolites, macromolecules, organelles, cell, synthetic markers, small molecules, organic polymers, hormones, peptides, antibodies, lipids, carbohydrates, inorganic and organic microparticles and nanoparticles, whole viruses, and any combination thereof.

First claim

The invention claimed is:

1. A system for identifying a particle based on a velocity of the particle, comprising:
a substrate;
a fluidic channel disposed on the substrate;
a voltage source;
one or more optical components;
one or more optical sensors;
one or more processors; and
a non-transitory computer-readable storage medium storing instructions which, when executed by the one or more processors, cause the system to perform the following:
generating, by the voltage source, an electrical field that induces electrophoretic or electroosmotic flow of a particle labeled with a fluorophore in a mixture through the channel;
illuminating the channel, by the one or more optical components, to form an excitation spot pattern that is incident on the channel, and that optically excites the particle as the particle flows past the excitation spot pattern;
detecting, by the one or more optical sensors, a plurality of signal spikes emitted by the particle in response to the particle being illuminated by the excitation spot pattern, wherein the plurality of signal spikes comprises a first fluorescence signal spike emitted by the particle in response to the particle being illuminated by the excitation spot pattern and a second fluorescence signal spike emitted by the particle in response to the particle being illuminated by the excitation spot pattern;
calculating a velocity of the particle based on the plurality of signal spikes; and
determining an identity of the particle based on the calculated velocity.

2. The system of claim 1 , wherein the instructions that cause the system to determine the identity of the particle comprise instructions for determining a molecular makeup of the particle.

3. The system of claim 1 , wherein the instructions that cause the system to determine the identity of the particle comprise instructions for determining a type of particle to which the particle corresponds.

4. The system of claim 1 , wherein the instructions that cause the system to determine the identity of the particle comprise instructions for determining whether the particle is a same type of particle as a second particle.

5. The system of claim 1 , wherein the instructions that cause the system to determine the identity of the particle comprise instructions for matching the calculated velocity to a previously-measured velocity of a second particle, wherein the previously-measured velocity is retrieved from a look-up table.

6. The system of claim 1 , wherein the instructions that cause the system to determine the identity of the particle comprise instructions for matching the calculated velocity of the particle to a theoretically calculated velocity for a type of particle.

7. The system of claim 1 , wherein the excitation spot pattern comprises a first excitation spot incident on the channel at a first location and a second excitation spot incident on the channel at a second location spaced apart from the first location by a spacing distance.

8. The system of claim 7 , wherein the instructions that cause the system to calculate the velocity of the particle comprise instructions for determining a time difference between a time at which the first fluorescence signal spike was detected and a time at which the second fluorescence signal spike was emitted, and calculating the velocity based on the time difference and the spacing distance.

9. The system of claim 7 , wherein the spacing distance is less than 20 cm.

10. The system of claim 1 , wherein the one or more optical components comprise a light source, and the instructions further cause the system to generate, by the light source, a spot pattern remote from the substrate.

11. The system of claim 1 , wherein the one or more optical components comprise one or more waveguides disposed on the substrate.

12. The system of claim 11 , wherein the one or more waveguides comprise one or more optical splitters, and the instructions further cause the system to split, by the one or more optical splitters, a single optical input into a plurality of optical outputs.

13. The system of claim 12 , wherein the one or more optical splitters comprise a plurality of optical splitters arranged in series, and the instructions further cause the system to create, by the plurality of optical splitters arranged in series, an excitation spot pattern having two or more excitation spots.

14. The system of claim 11 , wherein the one or more waveguides comprise a multi-mode interference waveguide that generates the excitation spot pattern as an interference-based spot pattern.

15. The system of claim 1 , wherein the plurality of signal spikes are detected as part of a raw signal, and the instructions further cause the system to:
generate a transformed signal by calculating a time-shifted product of the raw signal with the raw signal; and
calculate the velocity based on the transformed signal.

16. The system of claim 15 , wherein the instructions that cause the system to calculate the time-shifted product of the raw signal with the raw signal comprise instructions for calculating the product using one factor per excitation spots in the excitation spot pattern.

17. The system of claim 1 , wherein the instructions further cause the system to detect, by the one or more optical sensors, single particles comprising one or more of: an individual molecule, a particle included in single-cell lysate, an amino acid, a protein, a nucleic acid, a metabolite, a whole virus, a nucleic acid, and a metabolite.

18. The system of claim 1 , wherein the one or more optical sensors comprise an integrated optical sensor disposed on the substrate, and the instructions further cause the system to collect, by the integrated optical sensor, the plurality of signal spikes via planar beam paths.

19. The system of claim 18 , wherein the instructions further cause the system to collect, by the integrated optical sensor, the plurality of signal spikes via an integrated liquid core waveguide.

20. The system of claim 1 , wherein the channel is less than 5 m in length.

21. The system of claim 1 , wherein an internal volume of the fluidic channel excited by a spot of the spot pattern is less than 1 picoliter.

22. The system of claim 1 , wherein the instructions further cause the system to generate, by the voltage source, an electrical field for inducing electrophoretic and electroosmotic flow of the particle in the channel.

23. A system for separating particles based on individual particle velocity, comprising:
a substrate;
a fluidic channel disposed on the substrate;
a voltage source;
one or more optical components;
one or more optical sensors;
a particle separation control component;
one or more processors; and
a non-transitory computer-readable storage medium storing instructions which, when executed by the one or more processors, cause the system to perform the following:
generating, by the voltage source, an electrical field that induces electrophoretic or electroosmotic flow of a particle labeled with a fluorophore in a mixture through the channel;
illuminating the channel, by the one or more optical components to form an excitation spot pattern that is incident on the channel and that optically excites the particle as the particle flows past the excitation spot pattern;
detecting, by the one or more optical sensors, a plurality of signal spikes emitted by the particle in response to the particle being illuminated by the excitation spot pattern, wherein the plurali

Assignees

• Univ California
• Univ Brigham Young

Inventors

• Schmidt Holger
• Hawkins Aaron Roe

Classifications

• G01N2015/1027

  Determining speed or velocity of a particle · CPC title

• G01N15/149

  specially adapted for sorting particles, e.g. by their size or optical properties · CPC title

• B01L3/502761 Primary

  specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads or physically stretching molecules · CPC title

• B01L3/502715

  characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces · CPC title

• G01N15/1434 Primary

  Optical arrangements · CPC title