Single-Objective Light-Sheet Microscopy for Magnification Switching
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Conventional single-objective light-sheet fluorescence microscopes face limitations in achieving in-situ magnification switching and suffer from slow imaging speed due to strict optical parameter limitations and the need for perfect imaging data of the excitation plane.
Innovation Solution
A three-dimensional fluorescence imaging system using single-objective light-sheet microscopy that omits two remote imaging objectives for aberration correction, allowing for in-situ magnification switching and faster imaging by collecting projections of the excitation plane instead of perfect imaging data.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If three objective lenses are used for aberration-free remote focusing and oblique plane correction, then optical aberration is corrected and perfect imaging is realized, but the imaging system becomes complicated with low applicability
Solution Approach 1:
The patent removes the secondary and tertiary objective lenses from the conventional three-objective light sheet microscope, retaining only the primary objective lens. This extraction of unnecessary components simplifies the imaging system while maintaining the core functionality of light sheet illumination and fluorescence detection, directly resolving the contradiction between imaging quality and system complexity.
Solution Approach 2:
The single objective lens performs multiple functions: it serves as the illumination objective for generating the light sheet and as the detection objective for collecting fluorescence signals. This multi-functionality eliminates the need for separate secondary and tertiary objectives for oblique plane correction, simplifying the system while maintaining imaging capability.
2Measurement precision
If a high numerical aperture objective is used for single-objective illumination and detection, then optical aberration is reduced, but the field of view becomes smaller due to higher magnification
Solution Approach 1:
The patent introduces a magnification switching mechanism that allows dynamic adjustment between different magnification levels. The system can switch between high magnification (for detailed observation with smaller field of view) and low magnification (for broader field of view), enabling the user to optimize between resolution and field of view based on specific imaging needs.
Solution Approach 2:
The system changes the magnification parameter by switching between different objective lenses or lens configurations. This allows the numerical aperture and magnification to be adjusted according to the imaging requirements, resolving the fixed trade-off between field of view and resolution inherent in single-objective designs.
3Adaptability or versatility
If row by row exposure is used for in-situ magnification switching, then magnification switching is achieved, but imaging speed becomes slow
Solution Approach 1:
The patent employs a galvanometer to periodically scan the light sheet across the sample in a raster pattern, enabling rapid acquisition of multiple projection images. This periodic scanning action allows the system to capture comprehensive sample data much faster than sequential row-by-row exposure, maintaining magnification switching capability while dramatically improving imaging speed.
Solution Approach 2:
The system continuously scans the light sheet across the entire sample area in a systematic pattern, ensuring that useful imaging action occurs throughout the entire field of view simultaneously rather than progressing slowly row by row. This continuous scanning approach maintains high imaging speed while enabling magnification switching through rapid projection collection.
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 enables quick and convenient acquisition of three-dimensional structural information of biological samples with improved imaging speed and flexibility in magnification switching, expanding the application fields of light sheet fluorescence microscopes.
Implementation Method 1
a light sheet having an inclination angle α projected onto an excitation plane of a sample to excite fluorescence
Data Source
AI summary
A three-dimensional fluorescence imaging system, including an illumination objective, a scanning lens group, a galvanometer, an illumination module, and a detection module. The scanning lens group includes a first scanning lens and a second scanning lens; main optical axes of the first scanning lens and the second scanning lens are orthogonal and confocal; a common focus of the first scanning lens and the second scanning lens is disposed on a central part of the galvanometer. The illumination module is configured to generate an illumination laser beam; the illumination laser beam passes through the scanning lens group and enters a rear pupil plane of the illumination objective, forming a light sheet having an inclination angle α projected onto an excitation plane of a sample to excite fluorescence; the excitation plane is diagonally intersected with a main optical axis of the illumination objective. The detection module includes an area array detector.