Capillary Flow Cell Analyzer for Single-Molecule Detection

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Solution Overview

Problem

Current methods for detecting biological markers at low concentrations are limited in sensitivity and precision, making it difficult to diagnose early stages of conditions, monitor disease progression, and detect markers present in low amounts in biological samples.

Innovation Solution

A single molecule analyzer system that includes an electromagnetic radiation source, a capillary flow cell, a motive force, and a detector, using fluorescent moieties capable of emitting at least 200 photons when stimulated by a laser, allowing for the detection of markers over a wide dynamic range from femtomolar to picomolar concentrations.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If conventional detection methods are used, then device complexity is reduced, but measurement precision deteriorates due to inability to detect low concentrations of analytes

Engineering Contradiction:
Improvedetection sensitivityVSAvoidanalyzer system complexity
Core Design Contradiction:
Measurement precisionVSDevice complexity

Solution Approach 1:

The system segments the detection process into distinct functional modules: electromagnetic radiation source, capillary flow cell with interrogation space, motive force source, and detector. This modular segmentation allows each component to be optimized for its specific function while maintaining overall system sensitivity for detecting single molecules at femtomolar concentrations.

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

The patent introduces fluorescent moieties as intermediary substances that bind to target analytes and convert their presence into detectable electromagnetic signals. These fluorescent intermediaries amplify the detection signal, enabling single-molecule sensitivity while managing the complexity through a well-defined interaction mechanism.

Inventive Principle:
Principle #24Intermediary (Mediator)

2Measurement precision

If fluorescent moieties emitting 200+ photons are used, then measurement precision improves for low concentration detection, but use of energy increases due to laser stimulation requirements

Engineering Contradiction:
Improvesingle molecule detection accuracyVSAvoidlaser energy consumption
Core Design Contradiction:
Measurement precisionVSUse of energy by moving object

Solution Approach 1:

The system optimizes detection by carefully controlling laser power parameters and exposure duration to achieve sufficient photon emission (200+ photons per stimulus) while minimizing total energy consumption. The interrogation space geometry and fluorescent moiety characteristics are also tuned to maximize photon yield per unit energy input.

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The laser stimulation is applied in periodic pulses rather than continuous operation, allowing the fluorescent moieties to emit photons during each stimulus pulse while providing rest periods that reduce cumulative energy consumption and heat generation, thereby maintaining detection precision with lower overall energy use.

Inventive Principle:
Principle #19Periodic action

3Device complexity

If a single interrogation space is used, then device complexity is reduced, but measurement precision deteriorates due to limited dynamic range

Engineering Contradiction:
Improveinterrogation space configurationVSAvoidconcentration detection range
Core Design Contradiction:
Device complexityVSMeasurement precision

Solution Approach 1:

The system achieves a wide dynamic range (femtomolar to picomolar concentrations) in a single interrogation space by dynamically adjusting detection parameters such as laser power, integration time, and gain settings. This allows the same physical space to accurately detect analytes across five orders of magnitude in concentration without requiring multiple specialized chambers.

Inventive Principle:
Principle #15Dynamics

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

Enables highly sensitive detection and quantification of biological markers at extremely low concentrations, facilitating early diagnosis, disease monitoring, and treatment selection by overcoming the limitations of existing detection methods.

Implementation Method 1

a fluorescent moiety that is capable of emitting at least about 200 photons when simulated by a laser emitting light at the excitation wavelength of the moiety

Methodology Applied
Scientific EffectFluorescence: Fluorescence

Data Source

PatentEP3156799B1Analyzer and method for highly sensitive detection of analytes
Publication Date: 2024.01.24 NOVILUX LLC
  • EP3156799B1 patent drawingFigure 1A
  • EP3156799B1 patent drawingFigure 2A
  • EP3156799B1 patent drawingFigure 3A

AI summary

Disclosed is an analyzer for determining the presence, absence or amount of an analyte in a sample, wherein the analyzer comprises: (a) an electromagnetic radiation source (301) that emits electromagnetic radiation that defines an interrogation space comprising a focused spot of a beam from the electromagnetic radiation source (301); (b) a capillary flow cell (313) for passing a sample through the interrogation space; (c) a source of motive force for moving the sample in the capillary flow cell (313); (d) an electromagnetic radiation detector (309) operatively connected to the interrogation space that detects photons emitted from the interrogation space during a plurality of bins; and (e) a processor (310) operatively connected to the detector (309), wherein the processor determines the presence, absence or amount of an analyte in a sample by: (i) determining a threshold photon value corresponding to a background signal in the interrogation space, (ii) determining the presence of a photon emitting species comprising or corresponding to the analyte in the interrogation space in each bin of the plurality of bins by identifying bins having a photon value greater than the threshold value, (iii) comparing the number of bins having a photon value greater than the threshold value to a standard curve that is generated using one or more samples of known standard concentrations.