Waveguide-based detection system with scanning light source

• US 9,976,192 B2
• Filed: 10/03/2016
• Issued: 05/22/2018
• Est. Priority Date: 03/10/2006
• Status: Active Grant

• Current Assignee: LDIP, LLC
• Original Assignee: LDIP, LLC
• Inventors: Duer, Reuven, Chang, Yun-Pei, Shastry, Ashutosh
• Primary Examiner(s): AKANBI, ISIAKA O
• Application Number:
  US15/284,396
• Publication Number:
  US 20170023477A1
• Time in Patent Office: 596 Days
• Field of Search: 356311, 356432, 356445-446
• CPC Class Codes: B01L 2300/0636 Integrated biosensor, microarrays
• CPC Class Codes: B01L 2300/0654 Lenses; Optical fibres
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0819 Microarrays; Biochips
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: B01L 7/52 with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples amplification or hybridisation processes per se C12Q1/68; controlling sequential reactions for synthesis B01J19/0046
• CPC Class Codes: C12Q 1/6825 Nucleic acid detection involving sensors
• CPC Class Codes: C12Q 1/703 Viruses associated with AIDS
• CPC Class Codes: G01N 2021/6463 Optics
• CPC Class Codes: G01N 21/6428 Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 21/6452 Individual samples arranged in a regular 2D-array, e.g. multiwell plates
• CPC Class Codes: G01N 2201/0853 Movable fibre optical member, e.g. for scanning or selecting
• CPC Class Codes: G01N 2201/10 Scanning
• CPC Class Codes: G01N 33/54373 involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
• CPC Class Codes: G02B 6/4226 Positioning means for moving the elements into alignment, e.g. alignment screws, deformation of the mount

Abstract

The invention provides methods and devices for generating optical pulses in one or more waveguides using a spatially scanning light source. A detection system, methods of use thereof and kits for detecting a biologically active analyte molecule are also provided. The system includes a scanning light source, a substrate comprising a plurality of waveguides and a plurality of optical sensing sites in optical communication with one or more waveguide of the substrate, a detector that is coupled to and in optical communication with the substrate, and means for spatially translating a light beam emitted from said scanning light source such that the light beam is coupled to and in optical communication with the waveguides of the substrate at some point along its scanning path. The use of a scanning light source allows the coupling of light into the waveguides of the substrate in a simple and cost-effective manner.

Claims

1. A method of measuring a concentration of an analyte using a waveguide chip, the method comprising:

adding a known amount of one or more normalizing moieties to a sample;

adding a first detection molecule labelled with a first fluorescent tag to the sample, the first detection molecule specific to the analyte;

adding one or more secondary detection molecules, each secondary detection molecule labelled with a distinct secondary fluorescent tag, to the sample, the one or more secondary detection molecules specific to the one or more normalizing moieties, wherein the first fluorescent tag and the one or more secondary fluorescent tags emit at different fluorescence wavelengths;

introducing the sample to an array of sensing sites on the waveguide chip, each sensing site of the array having a first capture molecule specific to the analyte and one or more secondary capture molecules specific to the one or more normalizing moieties, wherein the first capture molecule and the one or more secondary capture molecules are both immobilized in each sensing site of the array at a predetermined ratio;

introducing a first light pulse into one or more excitation waveguides on the waveguide chip at a first predetermined interval, the one or more excitation waveguides in optical communication with the array of sensing sites, the first light pulse configured to excite the first fluorescent tag;

introducing one or more secondary light pulses into the one or more excitation waveguides on the waveguide chip at one or more secondary predetermined intervals, wherein the first light pulse and the one or more secondary light pulses have different wavelengths, the one or more secondary light pulses configured to excite the one or more secondary fluorescent tags;

measuring the fluorescence emitted by the first detection molecule and the one or more secondary detection molecules through a plurality of collection waveguides in optical communication with the array of sensing sites; and

normalizing the measured fluorescence emitted by the first detection molecule using the measured fluorescence emitted by the one or more secondary detection molecules.

2. The method of claim 1, wherein the step of normalizing the measured fluorescence emitted by the first detection molecule comprises dividing the measured fluorescence emitted by the first detection molecule by the measured fluorescence emitted by the one or more secondary detection molecules.

3. The method of claim 1, wherein the first detection molecule, the one or more secondary detection molecules, the first capture molecule, and the one or more secondary capture molecules are antibodies.

4. The method of claim 1, wherein the step of normalizing the measured fluorescence emitted by the first detection molecule further comprises generating one or more calibration curves from the measured fluorescence emitted by the one or more secondary detection molecules.

5. The method of claim 1, wherein the one or more normalizing moieties comprises at least a first normalizing moiety and a second normalizing moiety, wherein the amount of the first normalizing moiety added to the sample is greater than the amount of the second normalizing moiety added to the sample.

6. The method of claim 1, wherein the step of normalizing the measured fluorescence emitted by the first detection molecule comprises dividing the measured fluorescence emitted by the first detection molecule by a slope of the measured fluorescence emitted by the one or more secondary detection molecules.

7. The method of claim 1, wherein the step of normalizing the measured fluorescence emitted by the first detection molecule comprises dividing a slope of the measured fluorescence emitted by the first detection molecule by a slope of the measured fluorescence emitted by the one or more secondary detection molecules.