Microscope with Inclined Light Sheet and Microlens Array
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Solution Overview
Problem
Current microscopic imaging methods face challenges in efficiently capturing three-dimensionally resolved image data of large tissue volumes with high resolution and minimal background suppression, particularly for thick samples, due to limitations in frame rate, crosstalk, and the need for multiple objectives and complex apparatus.
Innovation Solution
A microscope with a controllable excitation beam path that generates an inclined light sheet using a scanning apparatus and a microlens array for light-field technology, allowing for adjustable illumination modes and full numerical aperture utilization, enabling efficient three-dimensional imaging with reduced background suppression and flexible objective use.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If confocal laser scanning microscopy is used to achieve high-contrast image stacks with background suppression, then measurement precision is improved, but productivity deteriorates due to very low frame rate from sequential scanning
Solution Approach 1:
The invention segments the illumination function by using multiple independent laser sources that can be activated simultaneously or sequentially, allowing parallel illumination of different regions while maintaining confocal background suppression for each region, thus improving frame rate without sacrificing measurement precision
Solution Approach 2:
The system dynamically switches between different illumination modes (single-point confocal, multi-point parallel, and light sheet modes) based on the imaging requirements, enabling adaptive adjustment of frame rate and resolution to optimize both productivity and measurement precision for different sample types and experimental conditions
2Length of stationary object
If multiphoton excitation is used to examine thicker samples, then the detection depth is improved, but productivity deteriorates due to sequential scanning
Solution Approach 1:
The invention divides the thick sample volume into multiple sub-volumes that can be illuminated and detected simultaneously using multiple laser foci at different depths, enabling parallel acquisition of multiple focal planes and significantly improving frame rate while maintaining deep tissue penetration through multiphoton excitation
Solution Approach 2:
The system employs periodic scanning of multiple laser foci through the sample volume in a coordinated manner, where each focus cycles through its designated depth range, allowing continuous high-speed acquisition of three-dimensional data from thick samples without the frame rate limitations of traditional sequential scanning
3Productivity
If spinning disk microscopy is used to scan multiple image points simultaneously, then productivity is improved, but measurement precision deteriorates due to crosstalk between adjacent pinholes in thick samples
Solution Approach 1:
The invention extracts the confocal pinhole filtering function and applies it independently to each laser focus through optical sectioning, eliminating the crosstalk problem inherent in spinning disk microscopy where pinholes are physically close together. Each virtual pinhole is optically isolated, allowing simultaneous multi-point illumination without background suppression degradation
Solution Approach 2:
The system introduces an intermediary optical sectioning mechanism that mediates between the multiple illumination points and the detector, using spatial filtering and confocal detection to prevent signal crosstalk while maintaining the high frame rate benefits of parallel illumination across thick sample volumes
4Productivity
If light sheet microscopy is used to achieve high frame rates, then productivity is improved, but measurement precision deteriorates due to limited optical sectioning and background suppression
Solution Approach 1:
The invention merges the advantages of light sheet microscopy (high frame rate, widefield detection) with confocal microscopy (optical sectioning, background suppression) by combining parallel multi-point illumination with virtual pinhole filtering for each focus, achieving both high productivity and high measurement precision simultaneously in thick sample imaging
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
This approach allows for high-speed imaging of large volumes with improved axial resolution and background discrimination, utilizing a single objective for both illumination and detection, and enabling flexible illumination patterns for various microscopy methods.
Implementation Method 1
a laser light source (2) for providing a laser light beam as excitation light
Implementation Method 2
guiding excitation light... a laser light beam is incident on a laser focus (18) in an objective pupil
Implementation Method 3
a scanning apparatus (6) for setting an angle of incidence of a focused laser light beam into a laser focus (18) in an objective pupil
Implementation Method 4
detected evaluation of captured detection light coming from a sample illuminated with the laser light
Data Source
AI summary
The invention relates to a microscope having an excitation beam path for guiding excitation light, having a laser light source for providing a laser light beam as excitation light and having a scanning apparatus for aligning and moving a focused laser light beam in the entrance pupil of an illumination objective; wherein the laser focus is directed into an entrance point that is offset with respect to the optical axis of the illumination objective; and also having a detection beam path for guiding detection light, comprising a microlens array having a focal plane for generating partial imaged presentations and a detector arranged in the focal plane of the microlens array for capturing the partial imaged presentations. In addition, an evaluation unit for evaluating the captured image signals of the detector in accordance with light-field technology is present. The invention additionally relates to a method for microscopic image generation.