Optical Super-Resolution Microscopy Using Annular Light and Varifocal Lens
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Solution Overview
Problem
Current optical super-resolution microscopic imaging technologies, such as STED, PALM/STORM, and SIM, face limitations in complexity, cost, light damage, repetitive processing, and balancing field of view with resolution, which restrict their applicability and efficiency.
Innovation Solution
An optical super-resolution microscopic imaging system utilizing a dichroic beamsplitter, focusing lens, confocal pinhole, varifocal lens system, and detector to form a single fluorescent excited light spot with a diameter smaller than the diffraction limit, achieving increased resolution without complex calculations or reconstructions, and maintaining high light collection efficiency.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If STED uses high intensity lost light to improve resolution, then resolution is improved, but light damage to biological sample increases
Solution Approach 1:
The patent changes the optical parameters by using annular parallel light with specific inner and outer diameters that satisfy particular mathematical relationships with the objective lens numerical aperture. This parameter optimization allows achieving super-resolution without requiring excessively high light intensity, thus reducing light damage while maintaining measurement precision.
Solution Approach 2:
The patent employs a varifocal lens system that can dynamically adjust the focal length to optimize the excitation light spot size. By dynamically controlling the optical path and light focusing, the system achieves high resolution with reduced light intensity requirements, thereby minimizing light damage to biological samples.
2Measurement precision
If PALM/STORM performs thousands of repetitions of activation-excitation-localization-bleaching, then super-resolution image is reconstructed, but time consumption increases
Solution Approach 1:
The patent replaces the complex multi-step chemical and computational reconstruction process of PALM/STORM with a direct optical imaging approach. By using specially designed annular parallel light excitation and confocal pinhole filtering, the system achieves super-resolution in a single shot or with minimal scanning, eliminating the need for thousands of repetitive cycles and complex image reconstruction algorithms.
3Measurement precision
If SIM uses CCD for fluorescence detection, then fluorescence information is received, but field of view and super-resolution cannot be balanced
Solution Approach 1:
The patent segments the detection approach by using a confocal pinhole to filter out-of-focus light, enabling optical sectioning. This allows the system to achieve super-resolution with an extended field of view by maintaining signal-to-noise ratio across larger areas, overcoming the CCD limitation where increasing field of view typically compromises resolution.
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 achieves a resolution increase of at least 1.6 times, providing a clear super-resolution image while minimizing light damage and maintaining maximum light collection efficiency, thus overcoming the limitations of existing technologies.
Implementation Method 1
a dichroic beamsplitter used for annular parallel light to transmit through
Implementation Method 2
a focusing lens used for converging the annular parallel light transmitted through the dichroic beamsplitter
Implementation Method 3
a confocal pinhole used for the annular parallel light after being converged to pass through so as to filter the annular parallel light
Implementation Method 4
a varifocal lens system used for collimating the annular parallel light passing through the confocal pinhole into excited annular parallel light
Implementation Method 5
forming a single fluorescent excited light spot with a diameter smaller than a diffraction limit of an objective lens on a sample positioned on a focal plane of the objective lens
Implementation Method 6
a detector used for receiving and processing fluorescence emitted by the excited sample
Data Source
AI summary
The invention provides an optical super-resolution microscopic imaging system comprising a dichroic beamsplitter for annular parallel light to transmit through; a focusing lens used for converging the annular parallel light transmitted through the dichroic beamsplitter; a confocal pinhole for the annular parallel light after being converged to pass through to filter the annular parallel light; a varifocal lens system for collimating the annular parallel light passing through the confocal pinhole into excited annular parallel light; and a detector for receiving and processing fluorescence emitted by the excited sample, the fluorescence emitted by the excited sample being returned by the same way, and the dichroic beamsplitter separating the fluorescence emitted by the sample from an annular parallel light path and turning the fluorescence to the detector to obtain a super-resolution image of the sample.