Controllable Phase Mask for Light Sheet Microscopy Intensity Homogenization
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Solution Overview
Problem
Current light sheet microscopy techniques face challenges in achieving homogeneous brightness in acquired images due to varying laser intensities and the inability to finely regulate illumination intensity, particularly in fluorescence microscopy, leading to mechanical complexity and high costs with existing solutions.
Innovation Solution
A method and apparatus for light sheet microscopy that control the phase angle deviation of a controllable phase mask to homogenize the intensity of illumination light across the light sheet's cross-sectional area, using a control unit to adjust the phase mask's phase pattern, allowing for continuous and fine adjustment of intensity without mechanical adaptations.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Shape
If a controllable phase mask is used to shape illumination light into a light sheet, then the light sheet can be formed with controlled phase patterns, but the illumination intensity becomes non-uniform (darker towards periphery) due to the Gaussian beam profile
Solution Approach 1:
The patent applies local quality by differentiating the phase angle deviation across different regions of the phase mask. Specifically, first regions (center) and second regions (periphery) have different phase angle deviations, allowing the illumination intensity to be locally adjusted to compensate for the Gaussian profile and achieve uniform intensity across the light sheet cross-section
Solution Approach 2:
The patent changes the phase parameter (phase angle deviation) of the phase mask to transform the illumination light. By controlling the phase mask to form phase patterns with spatially varying phase angle deviations, the Gaussian intensity profile is converted into a uniform intensity profile while maintaining the light sheet shape
2Illumination intensity
If acousto-optical elements or interchangeable attenuation filters are introduced to vary laser intensity, then the intensity can be regulated, but the mechanical-technical complexity and costs increase
Solution Approach 1:
The patent makes the phase mask multi-functional by using it for both light sheet shaping and intensity regulation. The same phase mask that shapes the light sheet can be controlled to provide different phase patterns that also control the illumination intensity, eliminating the need for separate acousto-optical elements or interchangeable filters
Solution Approach 2:
The patent merges the functions of light sheet generation and intensity regulation into a single phase mask component. By controlling the phase mask to form phase patterns with specific phase angle deviations, both the spatial shape and intensity of the illumination light are controlled simultaneously, reducing device complexity
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 enables the achievement of a substantially constant intensity profile over the light sheet's cross-sectional area, compensating for vignetting and allowing for precise intensity control, reducing mechanical complexity and costs while improving image brightness and fluorescence excitation.
Implementation Method 1
the controllable phase mask is controlled to form a phase pattern in which at least first regions and second regions are arranged in alternation, and wherein a greater phase angle deviation is impressed onto the illumination light in the first regions than in the second regions
Data Source
AI summary
The invention relates to a method and an apparatus for light sheet microscopy, in which a sample is illuminated with a light sheet and is observed with a microscope, wherein illumination light is shaped to form the light sheet using a controllable phase mask, wherein the controllable phase mask is controlled to form a phase pattern in which at least first regions and second regions are arranged in alternation, wherein a greater phase angle deviation is impressed on the illumination light in the first regions than in the second regions, and wherein at least the phase angle deviation of the first regions is controlled in a spatially dependent manner for influencing the intensity of the illumination light at different locations of a cross-sectional area of the light sheet. The invention additionally relates to a method and an apparatus for providing illumination light with variable intensity.