Droplet Sequencing Device Substrate Printing Fluorescence Detection

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Solution Overview

Problem

Current methods for sequencing nucleic acids using microdroplets are inefficient for analyzing nucleic acid fragments with significant nucleotide length, as they require dispersion in an immiscible carrier medium and lack suitable apparatus for precise droplet handling and fluorescence-based identification.

Innovation Solution

A droplet sequencing apparatus that prints droplets directly onto a planar substrate with droplet-receiving locations, using a printer head with nozzle groups to dispense single nucleotides and oligonucleotide probes, and employs fluorescence spectroscopy for identification, allowing for precise handling and analysis of long nucleic acid fragments.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If droplets are dispersed in an immiscible carrier medium for sequencing, then droplet manipulation is enabled, but the method becomes inefficient for analyzing nucleic acid fragments with significant nucleotide length

Engineering Contradiction:
Improvesequencing efficiencyVSAvoidcapability to analyze long nucleic acid fragments
Core Design Contradiction:
ProductivityVSAdaptability or versatility

Solution Approach 1:

The invention extracts the droplets from the immiscible carrier medium and deposits them directly onto a solid substrate. This removal of the carrier medium eliminates the limitations it imposes on analyzing long nucleic acid fragments while preserving the droplet-based manipulation advantages.

Inventive Principle:
Principle #2Taking out (Extraction)

Solution Approach 2:

The solid substrate acts as an intermediary between the droplet generation system and the detection system. It provides a stable platform that supports droplet containment and enables fluorescence detection without requiring the droplets to remain suspended in an immiscible carrier medium.

Inventive Principle:
Principle #24Intermediary (Mediator)

2Manufacturing precision

If a printer head with multiple nozzle groups is used to dispense droplets, then precise droplet placement is achieved, but device complexity increases

Engineering Contradiction:
Improvedroplet placement precisionVSAvoidapparatus structure
Core Design Contradiction:
Manufacturing precisionVSDevice complexity

Solution Approach 1:

The printer head is segmented into multiple nozzle groups, each responsible for dispensing specific types of droplets (e.g., nucleic acid-containing droplets, probe droplets, wash droplets). This segmentation enables precise control over droplet composition and placement while organizing the complexity into manageable functional units.

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

The invention transitions from single-dimensional droplet dispensing to multi-dimensional control by arranging nozzle groups in spatial configurations that enable precise two-dimensional addressing of droplet-receiving locations on the substrate, adding spatial organization to manage system complexity.

Inventive Principle:
Principle #17Another dimension (Dimensionality change)

3Adaptability or versatility

If droplets are printed directly onto a substrate, then handling of long nucleic acid fragments is improved, but the system requires new apparatus not previously used in sequencing

Engineering Contradiction:
Improvecapability to sequence long nucleic acid fragmentsVSAvoidnew apparatus requirements
Core Design Contradiction:
Adaptability or versatilityVSDevice complexity

Solution Approach 1:

The solid substrate serves multiple functions: it acts as a support structure for droplets, a platform for fluorescence detection, and a means for organizing and addressing individual nucleic acid fragments. This multi-functionality reduces the need for separate specialized components.

Inventive Principle:
Principle #6Universality (Multi-functionality)

Solution Approach 2:

The substrate-based droplet containment system enables self-aligned droplet placement and stable positioning without requiring active control mechanisms. The physical structure of the substrate provides inherent guidance and stabilization, reducing the complexity of control systems needed.

Inventive Principle:
Principle #25Self-service

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 effective sequencing of nucleic acid fragments exceeding 100 nucleotides by enhancing droplet formation and fluorescence detection, improving the efficiency and accuracy of nucleotide identification on a substrate with optimized droplet-receiving locations and illumination.

Implementation Method 1

at least one source of incident electromagnetic radiation adapted to illuminate the droplet-receiving locations and at least one photodetector adapted to detect fluorescence radiation emitted by each droplet-receiving location after illumination

Methodology Applied
Scientific EffectFluorescence: Fluorescence

Data Source

PatentEP3319729B1Droplet sequencing device
Publication Date: 2020.03.18 BASE4 INNOVATION LTD
  • EP3319729B1 patent drawingFigure 1
  • EP3319729B1 patent drawingFigure 2A~3
  • EP3319729B1 patent drawingFigure 4~5

AI summary

An apparatus for sequencing a nucleic acid by printing droplets at least some of which contain single nucleotides derived from the nucleic acid is provided. It is characterised by comprising; • a planar substrate having a face with droplet-receiving locations arranged in at least one track parallel to a first axis defining the substrate; · a printer head comprising a plurality of droplet-dispensing nozzle groups juxtaposed above the droplet-receiving locations so that each nozzle group can in turn dispense a droplet into the droplet-receiving locations; • a means for stepping the printer head along the first axis relative to the droplet-receiving locations; · a nucleotide-generating site upstream of the printer head by means of which an ordered stream of single nucleotides is created from the nucleic acid and delivered to a first droplet-dispensing nozzle group; • at least one source of incident electromagnetic radiation adapted to illuminate the droplet-receiving locations and · at least one photodetector to detect adapted to detect fluorescence radiation emitted by each droplet-receiving location after illumination.