Optical Imaging Equipment Using Microsphere Arrays for Super-Resolution
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Solution Overview
Problem
Conventional optical microscopes are limited by the optical diffraction limit, and existing super-resolution techniques require fluorescence labeling, which can interfere with samples and have limited imaging speed and field of view, while microsphere-based methods struggle with precise manipulation and large-area imaging.
Innovation Solution
An optical imaging equipment and method that uses a 3D electric sample table to move a sample relative to microspheres, allowing for precise control and scanning of multiple regions, enabling label-free, wide-field, far-field super-resolution imaging beyond the diffraction limit without damaging the sample.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If fluorescence super-resolution microscopy is used to achieve super-resolution imaging, then the resolution exceeds the optical diffraction limit, but the sample requires fluorescence labeling which interferes with the sample and some samples cannot be labeled
Solution Approach 1:
The patent extracts and eliminates the fluorescence labeling requirement from the imaging system. By using the photonic nanojet effect of transparent microspheres, the system achieves super-resolution imaging without requiring any fluorescent labels on the sample, thereby removing the harmful interference caused by labeling while preserving the super-resolution capability
Solution Approach 2:
The patent changes the fundamental imaging parameter from fluorescence-based detection to photonic nanojet-based detection. By utilizing the refractive index contrast and optical scattering properties of transparent microspheres rather than fluorescence emission, the system achieves label-free super-resolution imaging that is compatible with all sample types including those sensitive to labeling
2Measurement precision
If fluorescence super-resolution microscopy is used to achieve super-resolution imaging, then the resolution exceeds the optical diffraction limit, but the imaging speed is limited due to point-by-point mapping reconstruction
Solution Approach 1:
The patent replaces the mechanical point-by-point scanning and reconstruction process with a direct optical imaging approach. The photonic nanojet effect enables wide-field super-resolution imaging where the entire field of view is captured simultaneously through the objective lens, eliminating the time-consuming sequential point mapping and reconstruction process while maintaining super-resolution capability
3Measurement precision
If microspheres are used for super-resolution imaging, then the resolution can be less than 100nm, but the microsphere coverage area is very small and the observable field of view is limited
Solution Approach 1:
The patent merges multiple microspheres into a dense array configuration on the sample surface. By arranging numerous microspheres in close proximity, the individual small fields of view of each microsphere are combined to create a large cumulative field of view, enabling observation of extensive sample areas while maintaining the super-resolution capability of each microsphere
Solution Approach 2:
The patent transitions from single-microsphere imaging to a two-dimensional array of microspheres. By distributing microspheres across the sample surface in a planar arrangement, the system expands the observable area from the limited coverage of a single microsphere to a large field of view that encompasses multiple regions of interest simultaneously
4Manufacturing precision
If microspheres are manipulated by AFM tips or capillary to achieve precise positioning, then the microsphere can be located at any position, but the rigid connection causes damage to the sample or microsphere and breaks the tip during moving
Solution Approach 1:
The patent replaces the mechanical rigid connection method (AFM tips or capillary) with an optical manipulation approach using optical tweezers. The optical gradient forces generated by focused laser beams enable contactless manipulation and precise positioning of microspheres, eliminating the mechanical contact that causes damage to both the sample and microsphere while maintaining positioning precision
Solution Approach 2:
The patent introduces optical fields as an intermediary for microsphere manipulation. Instead of direct mechanical contact, the optical tweezers use light momentum transfer to levitate and position microspheres in three-dimensional space, providing a non-contact intermediary mechanism that avoids damage while achieving precise positioning control
5Area of stationary object
If a microsphere array is embedded in PDMS thin film to expand imaging area, then large area imaging is achieved, but the film thickness cannot be adjusted and Mosaic effect appears in the image
Solution Approach 1:
The patent introduces dynamic adjustability to the microsphere array system. By making the microsphere array structure adjustable in thickness and configuration, the system can be optimized for different imaging depths and sample types, eliminating the fixed thickness limitation of PDMS films and enabling precise focus control across the large field of view without Mosaic effects
Solution Approach 2:
The patent enables dynamic parameter adjustment of the microsphere array, specifically the thickness and spacing of microspheres. This allows the imaging system to be adapted to different sample depths and requirements, maintaining optimal focus and resolution across the entire large field of view while eliminating the Mosaic effect caused by fixed film thickness
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 high-resolution imaging exceeding the optical diffraction limit with a simple, cost-effective, and fast method suitable for living biological specimens, providing a direct wide-field optical image without the need for sample labeling, suitable for both transparent and opaque samples.
Implementation Method 1
micron-sized spheres of transparent media, or 'microspheres', placed on the surface of samples have a 'photonic nanojet effect'，meaning that it can transfer the evanescence wave of sample surface，which carrying fine structure information of the sample onto the distance
Implementation Method 2
it can transfer the evanescence wave of sample surface，which carrying fine structure information of the sample onto the distance
Implementation Method 3
the evanescence wave can be received by the optical objective lens placed in far field，which means the distance between the sample and the objective lens is much larger than the wavelength of light
Data Source
Figure 1
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Figure 4A~4B
AI summary
An optical imaging equipment and method. The optical imaging equipment comprises an optical microscope, an objective table, a light source module and an objective lens. The objective table is movable in the x-y plane, the light source module contains light illumination sources, and the objective lens is movable in the z-axis direction; a three dimensions (3D) electric sample table is fixed on the objective table, which is used for carrying a sample to be tested and driving the sample to move in 3D directions relative to the objective table, a microsphere is fixed on a transparent substrate; the objective lens, the microsphere and the sample to be tested are arranged in the z-axis direction in sequence, wherein, the transparent substrate along with the microsphere thereon can be moved to a first position and remain stationary relative to the objective table in the z-axis direction, the 3D electric sample table can adjust the sample to be tested with respect to the microsphere to an imaging plane which is parallel to the x-y plane and a first image is formed by the microsphere, the objective lens can be adjusted to a second position so that the objective lens can perform a secondary imaging of the first image to form a second image.