Objective-Lens Fluorescence Imaging With Offset Gratings
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Solution Overview
Problem
Existing high-throughput DNA sequencing systems face challenges in synchronizing illumination with image detection during scanning to minimize photodamage and ensure efficient imaging of genetic samples.
Innovation Solution
A system utilizing dual-surface imaging with offset gratings and planar waveguides, where laser beams of different wavelengths are directed through an objective lens to synchronize illumination and detection, allowing for low-power, low-background imaging with high-speed scanning.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional illumination methods (widefield illumination, confocal illumination) are used, then fluorescence imaging can be performed, but the axial resolution is limited and optical sectioning capability is insufficient
Solution Approach 1:
The patent implements nested illumination by placing a second illumination optical element (such as a lens or mirror) inside the first illumination optical element. The second element is positioned within the focal plane or optical path of the first element, creating a nested configuration that enables dual-stage illumination control for improved axial resolution while maintaining manageable system complexity
Solution Approach 2:
The patent introduces a second illumination optical element that operates in a different dimensional plane or optical path within the illumination system. This additional dimension of control allows independent optimization of axial and lateral illumination characteristics, achieving superior optical sectioning capability without proportionally increasing overall system complexity
2Measurement precision
If conventional illumination methods are used, then fluorescence imaging can be performed, but optical sectioning capability is limited
Solution Approach 1:
The nested configuration of illumination optical elements enables independent control of illumination at different depths. The second illumination element positioned within the first element's focal plane creates distinct illumination zones that provide superior optical sectioning capability while maintaining a compact and manageable system structure
Solution Approach 2:
The illumination system is segmented into multiple independent optical elements (first illumination optical element and second illumination optical element) that can be controlled separately. This segmentation allows independent optimization of illumination characteristics at different axial positions, achieving enhanced optical sectioning without requiring a completely complex reconfiguration of the entire system
3Measurement precision
If high numerical aperture objective lens is used, then lateral resolution is improved, but illumination uniformity and penetration depth are compromised
Solution Approach 1:
The patent applies local quality by using the nested illumination optical elements to create spatially varying illumination patterns that are optimized for different regions of the sample. The first and second illumination elements can be independently positioned and controlled to provide uniform illumination in specific axial zones, compensating for the illumination non-uniformity that typically occurs with high NA objective lenses
Solution Approach 2:
The system incorporates feedback mechanisms where the illumination optical elements can be dynamically adjusted based on the specific imaging requirements and sample characteristics. This feedback control allows optimization of illumination uniformity and penetration depth while maintaining the high lateral resolution capability of the high NA objective lens
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
Achieves up to 70 times reduction in laser power while maintaining signal-to-noise ratio, facilitating high-speed scanning and reducing photodamage, and supporting dual-color excitation with improved alignment tolerance and cost-effectiveness.
Implementation Method 1
an illumination optical element and an objective lens. The illumination optical element is configured to illuminate the sample with incident light. The objective lens is configured to focus the incident light onto the sample and to collect emitted light from the sample
Implementation Method 2
a fluorescence detector configured to detect the emitted light
Data Source
Figure 1
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AI summary
A system includes: an objective lens; a first light source to feed first illuminating light through the objective lens and into a flowcell (e.g., with a relatively thin film waveguide) to be installed in the system, the first illuminating light to be fed using a first grating on the flowcell; and a first image sensor to capture imaging light using the objective lens, wherein the first grating is positioned outside a field of view of the first image sensor. Dual-surface imaging can be performed. Flowcells with multiple swaths bounded by gratings can be used. An auto-alignment process can be performed.