Chromosomal Aberration Detection Using 3D Optical Sectioning
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Solution Overview
Problem
Current methods for detecting structural chromosome aberrations, such as inversions and translocations, face limitations in visualizing multiple colors simultaneously due to overlapping signals, leading to misdiagnosis and inability to reliably detect small inversions and insertions, especially in FISH and BrISH techniques.
Innovation Solution
A method using three differently labeled nucleic acid probes (A, B, and C) that flank breaking point regions, forming fusion signals A-B and C-C, which change to A-C and B-C upon chromosome aberrations, allowing for clear detection of inversions and translocations, particularly in small genomic sections, using combinations of fluorescent dyes or reporter molecules.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If multiple fluorescent dyes are used to detect multiple chromosome regions simultaneously, then the detection capability is improved, but the signals overlap and cannot be clearly differentiated
Solution Approach 1:
The patent transitions from 2D planar signal detection to 3D spatial resolution by using optical sectioning and z-stack imaging. This allows signals at different depths within the cell nucleus to be resolved separately, enabling clear differentiation of multiple fluorescent signals that would otherwise overlap in conventional 2D imaging.
Solution Approach 2:
The patent replaces conventional widefield fluorescence microscopy with confocal laser scanning microscopy, substituting a mechanical optical system that uses pinholes to eliminate out-of-focus light. This substitution enables optical sectioning and improves axial resolution, allowing clear separation of multiple fluorescent signals in three-dimensional space.
2Measurement precision
If only two or three colors are used for FISH detection, then signal differentiation is maintained, but the ability to detect multiple different chromosome regions is limited
Solution Approach 1:
The patent utilizes the third dimension (depth/z-axis) through optical sectioning to encode additional detection capacity. By resolving signals at different focal planes, the system can detect more than three chromosome regions simultaneously without signal overlap, effectively adding a spatial dimension to the detection capacity.
Solution Approach 2:
The patent segments the cell nucleus into multiple optical sections along the z-axis. Each section can contain distinct fluorescent signals at different depths, allowing multiple chromosome regions to be detected and differentiated by their spatial positions in three-dimensional space rather than relying solely on color differentiation.
3Ease of operation
If conventional FISH or BrISH methods are used, then the detection process is simple, but small inversions and insertions cannot be reliably detected
Solution Approach 1:
The patent replaces conventional fluorescence microscopy with confocal laser scanning microscopy, which provides optical sectioning and three-dimensional imaging capabilities. This substitution enables reliable detection of small inversions and insertions by resolving fine spatial details and distinguishing true signals from background noise through optical sectioning.
Solution Approach 2:
The patent introduces three-dimensional spatial resolution through z-stack imaging and optical sectioning. This additional dimensional information allows for more accurate detection and characterization of small chromosomal abnormalities by analyzing signal positions and patterns in 3D space, improving detection accuracy while maintaining operational feasibility.
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 reliable detection of structural chromosome aberrations, including small inversions and translocations, by forming distinct new fusion signals, improving diagnostic accuracy and differentiating between inversion and translocation events.
Implementation Method 1
in situ hybridization (ISH) using fluorescence-labeled (fluorescence ISH (FISH)) nucleic acid fragments, so-called probes
Data Source
AI summary
The invention relates to a method for detecting several different chromosomes or DNA regions in a cell in order to provide evidence for structural chromosomal aberrations, wherein the chromosomal aberrations have at least two breaking point regions within a chromosome, on the basis of directly or indirectly labeled nucleic acid fragments (probes), wherein: a first probe labeled with label A (probe A) and a second probe labeled with label B (probe B) flank a breaking point region 1, and form the fusion signals A-B; and two probes, a third and a fourth, each labeled with a label C (probes C), flank a breaking point region 2, and form the fusion signals C-C, wherein the above-mentioned fusion signals change in the event of a chromosomal aberration to fusion signals A-C and to fusion signals B-C.