Microscopy Illumination Lattice for Faster High-Resolution Confocal Imaging
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Solution Overview
Problem
Conventional structured illumination microscopy (SIM) methods require complex sample acquisition with multiple orientations and rotations of 1D lattice illumination, leading to lengthy image reconstruction times and suboptimal signal-to-noise ratio (SNR) due to crosstalk and out-of-focus light suppression challenges.
Innovation Solution
Employing asymmetrical 2D Bravais lattices with a longer first primitive vector and a shorter second primitive vector in combination with scanned line illumination, using an illumination modulator to create a light strip that minimizes crosstalk and allows for efficient line-confocal detection, thereby improving resolution and SNR.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If 1D line lattice illumination with multiple orientations and rotations is used to improve resolution and confocality, then image quality improves, but acquisition time increases significantly and device complexity increases
Solution Approach 1:
The patent transitions from 1D line lattice illumination to 2D lattice illumination, adding a spatial dimension to the illumination pattern. This allows simultaneous sampling of multiple spatial frequencies in both x and y directions within a single acquisition, achieving high resolution without requiring multiple rotational acquisitions. The 2D lattice pattern with primitive vectors a and b illuminates the sample in a grid arrangement, enabling parallel information gathering across the field of view.
Solution Approach 2:
The illumination pattern is segmented into discrete lattice points arranged in a 2D grid, where each point contributes to the structured illumination. This segmentation allows the system to sample different spatial frequencies at each lattice point position, and by combining signals from multiple lattice points simultaneously, the system achieves comprehensive frequency sampling without temporal sequencing of rotations.
2Object-affected harmful factors
If 1D line lattice illumination with multiple orientations is used to improve confocality, then out-of-focus light suppression improves, but device complexity and acquisition time increase
Solution Approach 1:
By extending the illumination pattern from 1D to 2D lattice structure, the system achieves confocality through spatial distribution of illumination points rather than temporal sequencing of rotational acquisitions. The 2D lattice with primitive vectors a and b creates a grid of illumination points that simultaneously suppress out-of-focus light across the entire field of view in a single acquisition plane.
3Measurement precision
If symmetrical 2D patterns with equal primitive vectors are used to improve resolution, then resolution improves, but crosstalk between illumination points increases
Solution Approach 1:
The patent employs asymmetrical 2D lattice patterns where the primitive vectors a and b have different lengths. This asymmetry strategically positions illumination points at different spatial separations, creating unequal distances between adjacent lattice points in different directions. The asymmetric arrangement reduces overlap of point spread functions from neighboring illumination points, thereby minimizing crosstalk while maintaining high spatial frequency sampling capability for resolution enhancement.
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 significantly reduces acquisition times and enhances image contrast by minimizing crosstalk and out-of-focus light, enabling faster and more efficient high-resolution 3D microscopy with improved SNR.
Implementation Method 1
an objective (20) is disposed in the illuminating beam path (24) in order to focus the illuminating light (27a) into the sample (32)
Implementation Method 2
an illumination modulator (17) is disposed in the illuminating beam path (24), optics (28) disposed in the illuminating beam path (24) together with the illumination modulator (17) produce a light strip (3) of the illuminating light (27a) in the sample (32) at the focus of the objective (20)
Implementation Method 3
a scanning unit (29) disposed in the illuminating beam path (24) is controlled in order to displace the light strip (3) of points of illumination (8, 11) in the sample (32) in a scanning direction (4)
Implementation Method 4
a detector (31) with an adjustable region of interest (12) detects radiation (25a) from the sample (32)
Data Source
Figure 1
Figure 2A~2C
Figure 3
AI summary
The invention concerns a microscopy method and a microscopy apparatus (100), with an illumination modulator (17) which is configured so as to produce a plurality of points of illumination (8, 11) during a scan of a light strip (3) of the illuminating light (27a) with an illumination modulator (17a), the points being in the form of an asymmetrical 2D Bravais lattice (G4, G5, G6) with a first, longer primitive vector (a) and a second, shorter primitive vector (b) and wherein the projection of the first primitive vector (a) onto the axial direction (3a) of the light strip (3) is longer than the projection of the second primitive vector (b) onto the axial direction (3a) of the light strip (3). In combination with a ROI of a detector synchronised to the illumination line, a more rapid confocal acquisition method is obtained with a better signal-to-noise ratio.