Hexagonal Fiducial Layout for Sequencing Image Registration

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Solution Overview

Problem

As the density of features in nucleic acid arrays increases, accurate detection becomes problematic due to challenges in locating sites, aligning them in successive cycles, and avoiding issues that confuse comparison of successive images or image data.

Innovation Solution

The use of fiducial features arranged in a hexagonal pattern, which include at least one depression with objects comprising fluorescent material, provides a solution by allowing for the registration of successive images and transformation of image data into sequence data.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If the density of features in the array increases, then the productivity and information capacity of the array improves, but the measurement precision and detection accuracy deteriorate

Engineering Contradiction:
Improvesequencing capacityVSAvoiddetection accuracy
Core Design Contradiction:
ProductivityVSMeasurement precision

Solution Approach 1:

Fiducial features are introduced as intermediary reference elements that do not participate in the biological sequencing function but mediate the detection process. These fiducial features provide stable reference points for image registration and alignment, enabling accurate detection of high-density biological features by serving as a mediator between the imaging system and the dense feature array

Inventive Principle:
Principle #24Intermediary (Mediator)

Solution Approach 2:

The fiducial features serve multiple functions: they provide reference points for image registration, enable alignment of successive images, assist in determining imaging parameters, and help correct for drift or distortion. This multi-functionality allows a single feature type to address multiple detection challenges simultaneously

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

2Quantity of substance

If the density of features in the array increases, then more genetic information can be analyzed, but the difficulty of detecting and measuring individual sites increases

Engineering Contradiction:
Improvenumber of sitesVSAvoidsite location difficulty
Core Design Contradiction:
Quantity of substanceVSDifficulty of detecting and measuring

Solution Approach 1:

The array is segmented into functional regions: biological feature sites for sequencing and fiducial feature sites for reference. This segmentation allows the detection system to distinguish between sites requiring biological analysis and sites providing geometric reference, simplifying the detection process by separating measurement functions from biological functions

Inventive Principle:
Principle #1Segmentation

3Area of stationary object

If the overall size of the array expands, then more samples can be processed, but the alignment and indexing of sites in successive cycles becomes more problematic

Engineering Contradiction:
Improvearray sizeVSAvoidalignment precision
Core Design Contradiction:
Area of stationary objectVSManufacturing precision

Solution Approach 1:

Fiducial features are pre-positioned at known locations throughout the array before imaging begins. These pre-established reference points enable the system to perform preliminary alignment and establish coordinate transformations before actual biological feature detection, facilitating accurate indexing across large array areas

Inventive Principle:
Principle #10Preliminary action

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

This approach enhances the accuracy of sequence analysis by improving the registration of images and data, thereby addressing the challenges associated with high-density feature arrays.

Implementation Method 1

The plurality of spots may comprise at least one reference spot arranged for encoding identification information for identification of said two-dimensional sensor-array. The at least one reference spot may comprise a fluorescent material.

Methodology Applied
Scientific EffectFluorescence: Fluorescence

Data Source

PatentEP3577441B1System and method with fiducial features in a hexagonal arrangement
Publication Date: 2025.05.21 ILLUMINA INC
  • EP3577441B1 patent drawingFigure 1
  • EP3577441B1 patent drawingFigure 2~4
  • EP3577441B1 patent drawingFigure 5~7

AI summary

Fiducial markers are provided on patterned arrays of the type that may be used for molecular analysis, such as sequencing. The fiducials may have configurations and layouts that enhance their detection in image or detection data, that facilitate or improve processing, that provide encoding of useful information, and so forth. Examples of the fiducials may include offset layouts that may be useful in detecting the fiducials in different types and approaches in imaging, and that may help to distinguish regions of the array from one another in image data.