Multi-Depth Confocal Imaging Without High-NA Immersion Lenses
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Solution Overview
Problem
Conventional fluorescent microscopy techniques face challenges such as high photodamage, complex workflows, and space constraints due to the need for high-NA immersion objective lenses, limiting throughput and applicability to certain samples.
Innovation Solution
A multi-depth confocal imaging system that uses a combination of oblique-plane and confocal excitation, eliminating the need for high-NA immersion lenses by simultaneously imaging multiple focal depths with a synergistic approach, incorporating fiber optics and oblique-oriented image sensors to achieve high-throughput imaging.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If high-NA immersion objective lenses are used to achieve high optical resolution, then imaging resolution is improved, but device complexity and space requirements increase
Solution Approach 1:
The patent segments the imaging process by using multiple low-NA objectives instead of a single high-NA immersion objective. Each objective captures images at different focal depths, and computational methods combine these segments to achieve high-resolution 3D imaging without requiring complex immersion lens systems
Solution Approach 2:
The patent transitions from 2D planar imaging with high-NA lenses to 3D volumetric imaging using multiple low-NA objectives at different focal depths. By adding the depth dimension through multi-focal plane capture and computational reconstruction, the system achieves high resolution without the complexity of immersion optics
2Measurement precision
If conventional fluorescent microscopy is used to capture high-resolution images, then imaging quality is improved, but photodamage to the sample increases
Solution Approach 1:
The patent divides the total imaging task across multiple low-NA objectives capturing different focal depths. This segmentation allows lower illumination intensity per objective while maintaining overall image quality, thereby reducing photodamage to the sample compared to using a single high-NA objective requiring high intensity
Solution Approach 2:
The patent creates multiple copies of the sample at different focal depths using multiple objectives, then computationally reconstructs the high-resolution 3D image from these copies. This copying approach enables high-quality imaging with distributed, lower-intensity illumination rather than concentrated high-intensity light
3Productivity
If Light Sheet Fluorescence Microscopy is used for rapid 3D imaging, then imaging speed is improved, but system complexity and alignment difficulty increase
Solution Approach 1:
The patent merges multiple conventional microscopy objectives into a unified multi-depth imaging system. By combining the simplicity of standard objectives with computational multi-focal plane processing, the system achieves rapid 3D imaging capabilities similar to light-sheet microscopy but with lower operational complexity and easier alignment
Solution Approach 2:
The patent replaces complex mechanical alignment systems (like those in light-sheet microscopy requiring precise orthogonal objective positioning) with a computational approach. Multiple objectives at fixed focal depths capture images simultaneously, and software reconstructs the 3D volume, substituting mechanical complexity with computational processing
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
The system significantly increases imaging throughput by concurrently capturing multiple focal depths without requiring high-NA immersion lenses, addressing space constraints and reducing photodamage, thus enhancing the versatility and efficiency of fluorescent microscopy.
Implementation Method 1
illuminating a sample at a plurality of depths and detecting light from the sample (e.g., fluorescent excitation events, scattered light, transmitted light, or reflected light)
Implementation Method 2
the excitation beams are focused into a sample at a first plurality of focus depths (e.g., two or more different depths, such as 2-10 different depths, 4-12 different depths, 5-8 different depths) along an excitation direction through the objective lens
Implementation Method 3
an image sensor that receives emissions (e.g., light beams, such as excited emissions, fluorescent excitation emissions, scattered light emissions, transmitted light emissions, or reflected light emissions) from the sample
Data Source
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AI summary
A multi-depth confocal imaging system includes at least one light source configured to provide excitation beams and an objective lens. The excitation beams are focused into a sample at a first plurality of focus depths along an excitation direction through the objective lens. An image sensor receives emissions from the sample via the objective lens, wherein the emissions define foci relative to the image sensor at a second plurality of focus depths.