Non-Point Optical Sectioning for 3D Sperm Quality Scoring
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Solution Overview
Problem
Existing sperm quality determination methods are limited by 2D confinement, restricted field of view, and inability to simultaneously image multiple freely swimming spermatozoa, lacking the resolution to reconstruct morphological details and dynamics effectively.
Innovation Solution
A method utilizing non-point scanning optical sectioning microscopy for sequential acquisition of volumetric image groups, extracting three-dimensional morphological and dynamics features of spermatozoa, and determining a quality score based on statistical comparisons with benchmarks.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If a high magnification objective is used to resolve morphological details of the head and flagellum, then measurement precision is improved, but the field of view is reduced allowing only a few spermatozoa to be visualized
Solution Approach 1:
The system divides the observation space into multiple focal planes along the z-axis, sequentially imaging different depth layers. This segmentation allows high-magnification objectives to maintain detailed morphological resolution while the systematic scanning across multiple planes enables comprehensive coverage of multiple spermatozoa throughout the sample volume.
Solution Approach 2:
The system transitions from 2D planar imaging to 3D volumetric imaging by adding the z-axis dimension through optical sectioning. This dimensional extension allows simultaneous visualization of multiple spermatozoa at different depths while maintaining high lateral resolution through the use of high magnification objectives, effectively resolving the field of view limitation.
2Quantity of substance
If a low magnification objective is used to enable a large field of view and imaging of multiple spermatozoa, then quantity of substance is improved, but measurement precision deteriorates with limited resolution that only allows head tracking
Solution Approach 1:
The system segments the volumetric sample into multiple optical sections at different z-positions. By sequentially imaging these sections and reconstructing them computationally, the system achieves high-resolution morphological details of multiple spermatozoa throughout the volume, overcoming the resolution limitations of low magnification objectives.
Solution Approach 2:
The system adds the z-axis dimension to enable volumetric imaging of multiple spermatozoa. Through optical sectioning and 3D reconstruction, the system recovers high-resolution morphological details that would be unavailable in 2D imaging, allowing simultaneous visualization of multiple cells with detailed structural information.
3Ease of operation
If CASA systems constrain spermatozoa movement in a 2D plane, then ease of operation is improved with simplified imaging, but reliability deteriorates as spermatozoa are not freely moving in a volume
Solution Approach 1:
The system transitions from 2D planar confinement to 3D volumetric imaging, allowing spermatozoa to swim freely in three dimensions. The optical sectioning technique captures images at multiple z-positions, reconstructing the complete 3D trajectories and natural swimming behaviors of spermatozoa without artificial planar constraints.
Solution Approach 2:
The system creates a virtual 3D copy of the sperm sample volume through computational reconstruction of optical sections. This digital replica preserves the natural 3D swimming environment and trajectories, allowing analysis of authentic biological behavior without physical constraints on sperm movement.
4Measurement precision
If piezoelectric systems use a piezoelectric stage to rapidly adjust the position of a microscope objective, then measurement precision is improved for 3D tracking, but device complexity increases and the field of view remains limited
Solution Approach 1:
The system replaces the mechanical piezoelectric stage with an optical sectioning approach. Instead of physically moving the objective through the sample volume, the system uses optical methods to sequentially image different z-planes, achieving 3D tracking capability without the mechanical complexity and field of view limitations of piezoelectric positioning systems.
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 simultaneous imaging and characterization of multiple freely moving spermatozoa, providing reliable quality scores through decoupled depth of field and detection objective, allowing natural swimming conditions for accurate 3D analysis.
Implementation Method 1
locating and tracking image data representative of at least one spermatozoon in a plurality of sequentially acquired volumetric groups of images of a sample that includes a plurality of freely swimming spermatozoa, and wherein the sequential acquisition has been carried out by means of non-point scanning optical sectioning microscopy
Data Source
AI summary
The present invention relates to a method for sperm quality determination, comprising:a) locating and tracking image data of a spermatozoon in a plurality of sequentially acquired volumetric groups of images of a sample that includes a plurality of freely swimming spermatozoa, wherein the sequential acquisition has been carried out by means of non-point scanning optical sectioning microscopy;b) extracting features from the image data, the extracted features being three-dimensional morphological and dynamics features of the head and flagellum of the spermatozoon;c) determining a quality score for the spermatozoon based on the extracted three-dimensional features, applying statistics, and conducting a comparison with benchmarks referring to spermatozoa morphology and dynamics; andd) providing the quality score.The present invention also relates to a computer program and a system implementing the method of the invention.