Light Sheet Microscope Z-Resolution via Thickness Optimization
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Solution Overview
Problem
Conventional light sheet microscopes face challenges in achieving high z-resolution and accuracy in three-dimensional information acquisition due to the trade-off between light sheet thickness and numerical aperture, leading to increased z-stack images and reduced image contrast.
Innovation Solution
A three-dimensional information obtaining apparatus that illuminates a sample with a light sheet of controlled thickness, capturing images from multiple directions and using correlation measurement methods to calculate three-dimensional information, with the light sheet thickness optimized to balance resolution and image contrast, adhering to specific conditional expressions for pixel pitch, wavelength, magnification, and numerical aperture.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If the light sheet thickness is reduced to improve z-resolution, then the measurement precision improves, but the image contrast deteriorates and the number of z-stack images increases
Solution Approach 1:
The patent changes the parameter of light sheet thickness to a specific range (D ≤ 8·PP/β/NA when PP > λ·β/(4·NA)) that balances both z-resolution and image contrast, rather than simply minimizing thickness. This parameter optimization resolves the contradiction by finding the optimal thickness value that satisfies both requirements simultaneously.
2Measurement precision
If the light sheet thickness is reduced to improve z-resolution, then the measurement precision improves, but the image acquisition time increases
Solution Approach 1:
By optimizing the light sheet thickness to a specific range rather than minimizing it, the patent reduces the number of z-stack images needed while maintaining high z-resolution. This parameter optimization resolves the contradiction by achieving both high precision and efficient data collection.
3Measurement precision
If the numerical aperture is increased to improve resolving power, then the measurement precision improves, but the illumination area decreases
Solution Approach 1:
The patent optimizes the light sheet thickness parameter in relation to the numerical aperture using the conditional expression D ≤ 8·PP/β/NA. This allows maintaining a practical illumination area while achieving high resolving power through coordinated parameter optimization rather than simply increasing NA.
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 rapid acquisition of highly accurate three-dimensional information with improved z-resolution and reduced image acquisition time, while maintaining image contrast and accuracy, even when the light sheet is thicker, allowing for wider illumination and more efficient data collection.
Implementation Method 1
an illumination optical system that illuminates a sample with a light sheet
Implementation Method 2
an observation optical system that forms, on the two-dimensional imaging element, a plurality of optical images of the sample observed from a plurality of different directions
Implementation Method 3
an imaging device that has a two-dimensional imaging element and that captures an image of the sample illuminated by the illumination optical system
Data Source
AI summary
A three-dimensional information obtaining apparatus includes an illumination optical system that illuminates a sample with a light sheet, an imaging device that has a two-dimensional imaging element and that captures an image of the sample illuminated by the illumination optical system, an observation optical system that forms, on the two-dimensional imaging element, a plurality of optical images of the sample observed from a plurality of different directions, and an arithmetic device that calculates three-dimensional information of the sample from a plurality of pieces of image data of the sample. A thickness D of the light sheet satisfies D≤8·PP/β/NA when PP>λ·β/(4·NA) is satisfied, where PP is a pixel pitch of the two-dimensional imaging element, λ is a wavelength of observation light, β is a magnification of the plurality of optical images, and NA is a numerical aperture on an object side of the observation optical system.