Microscope with Color Splitter and Cylindrical Elements for Light-Field Imaging
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Solution Overview
Problem
Current microscopy methods for capturing three-dimensional sample volumes, particularly in neural networks, face challenges such as slow data capture, susceptibility to artifacts, low parallelization, and reduced contrast due to limitations in light-sheet illumination and detection techniques.
Innovation Solution
A microscope system with a flexible operating mode that uses a color splitter and cylindrical optical elements to generate and control a thick light sheet for improved parallelization and contrast, allowing for light-field microscopy with enhanced resolution and reduced background radiation, and switching between light-sheet and point-type illumination modes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If light-sheet illumination is used to illuminate a large volume, then parallelization is improved, but contrast is reduced due to strong background radiation from out-of-focus structures
Solution Approach 1:
The detection path is segmented into multiple channels: a light-sheet detection path for capturing in-focus fluorescence and a widefield detection path for capturing out-of-focus background radiation. These segmented paths allow independent processing and combination of signals, enabling both high parallelization from light-sheet illumination and improved contrast by separating background radiation
Solution Approach 2:
A beam splitter is introduced as an intermediary element that divides the fluorescence radiation from the sample into two separate detection paths. This intermediary enables simultaneous light-sheet and widefield detection by directing different portions of the light to different detectors, resolving the contradiction between parallelization and contrast
2Productivity
If scanning speed is increased to capture neural signals with high temporal resolution, then productivity is improved, but luminous power must be increased which saturates fluorescent markers and damages the sample
Solution Approach 1:
The scanning system is segmented into multiple independent scanning beams that operate in parallel. Instead of sequentially scanning one beam through the entire volume, multiple beams simultaneously scan different regions, achieving high temporal resolution without increasing the power of individual beams, thus avoiding sample damage
Solution Approach 2:
Multiple scanning beams cover different regions of the sample simultaneously, providing partial coverage that collectively achieves complete volumetric imaging. This distributed approach allows faster data capture without concentrating excessive power in a single location, preventing fluorescent marker saturation and sample damage
3Measurement precision
If a microlens array is used in light-field microscopy to capture large volumes with improved resolution, then measurement precision is improved, but only a part of the aperture is used by each microlens group reducing efficiency
Solution Approach 1:
The system transitions from traditional light-field microscopy that captures spatial information to a configuration that also captures angular information through the microlens array arrangement. This dimensional change in information capture allows full aperture utilization while maintaining high resolution through computational processing of the multi-dimensional data
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
Enables fast and high-resolution imaging of large volumes with improved contrast and reduced sample damage, allowing for comprehensive capture of neural signals with increased information content by combining light-field and confocal detection methods.
Implementation Method 1
a color splitter (8), which is reflective to the excitation radiation and through the effect of which a beam of the excitation radiation is reflected along the further excitation beam path
Implementation Method 2
a first cylindrical optical element (5.1) which limits a cross section of the beam of the excitation radiation transversely to the propagation direction of the excitation radiation
Implementation Method 3
an objective (11) for illuminating a sample (13) with the excitation radiation and for capturing detection radiation brought about by means of the excitation radiation
Implementation Method 4
a first detector (17) having an upstream microlens array (18), with the result that, in addition to a piece of spatial information of the origin of the detection radiation, a plurality of pieces of angle information of the detection radiation can also be captured
Data Source
AI summary
A microscope, which includes a color splitter that is reflective to excitation radiation, can switch between a first and a second operating mode. A first apparatus can introduce a first cylindrical optical element into the excitation beam path between a light source and the color splitter, when the microscope is in the first operating mode, and a second apparatus can introduce a second cylindrical optical element into the excitation beam path between the color splitter and a scanning apparatus.