Variable-Illumination Imaging for Adjustable Optical Sectioning
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Conventional confocal microscopy is limited by a fixed pinhole size that restricts optical sectioning versatility and requires multiple scans for varying sectioning levels, leading to inefficiencies and potential sample degradation.
Innovation Solution
A method and device that utilize a focused illumination beam with variable intensity at multiple lateral positions to construct optically-sectioned images, allowing for adjustable sectioning through post-processing without hardware changes, using a combination of structured illumination and statistical thresholding.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If a fixed pinhole size is used in confocal microscopy, then the optical sectioning level is fixed, but the versatility of the microscope is limited
Solution Approach 1:
The patent applies the dynamics principle by replacing the fixed pinhole with a variable aperture that can dynamically adjust its size. The aperture is controlled by a piezoelectrically actuated membrane that can change the opening size in real-time, allowing the system to adapt the optical sectioning level to different imaging requirements without physical reconfiguration.
Solution Approach 2:
The patent implements parameter changes by varying the aperture size parameter to control the degree of optical sectioning. By changing the aperture diameter from fully open to fully closed states, the system can continuously adjust the sectioning level, transforming a fixed-parameter system into a variable-parameter system that adapts to different biological samples and imaging conditions.
2Measurement precision
If the pinhole size is reduced to improve optical sectioning, then the optical contrast improves, but the amount of collected light from the in-focus plane is reduced
Solution Approach 1:
The dynamic aperture allows the system to optimize the balance between optical contrast and light collection by adjusting the opening size in real-time. During imaging, the aperture can be opened wider to maximize light collection, then narrowed during post-processing to enhance contrast, eliminating the need to choose a fixed compromise setting.
Solution Approach 2:
The system performs preliminary imaging with the aperture in an open state to capture maximum light, then applies computational sectioning algorithms in post-processing to achieve the desired optical contrast. This preliminary action of capturing all available light before selective processing resolves the contradiction between light collection and contrast enhancement.
3Adaptability or versatility
If multiple scans are performed to achieve varying sectioning levels, then optical sectioning versatility is achieved, but the time required and sample degradation increase
Solution Approach 1:
The system performs a single preliminary scan with the aperture fully open to capture all necessary optical information. The varying sectioning levels are then achieved through post-processing algorithms that computationally reconstruct images at different focal planes, eliminating the need for multiple physical scans and reducing both time and sample exposure.
Solution Approach 2:
Instead of performing multiple physical scans to create different sectioning levels, the system creates computational copies of the same raw data through algorithmic processing. The post-processing algorithms generate multiple virtual sectioned images from a single scan, achieving versatility without repeated physical measurement and minimizing sample degradation.
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 continuous adjustment of optical sectioning without re-scanning, reducing hardware requirements and minimizing sample degradation, while maintaining image quality and versatility.
Implementation Method 1
providing an illumination beam through an imaging lens such that the illumination beam is focused at a focal plane of the imaging lens
Implementation Method 2
detecting, using a detector, signals collected via the imaging lens
Data Source
Figure 1
Figure 2
Figure 3a
AI summary
A method is presented for obtaining an optically-sectioned image of a sample. The method comprises: providing an illumination beam through an imaging lens such that the illumination beam is focused at a focal plane of the imaging lens; obtaining a plurality of images of the sample. Obtaining comprises providing the illumination beam at a plurality of lateral positions on the focal plane and obtaining each image at each lateral position of the illumination beam, such that an intensity of the illumination beam on a portion of the sample at the focal plane varies for each of the plurality of lateral positions. The method further comprises detecting, using a detector, signals collected via the imaging lens; and constructing the optically-sectioned image based on the plurality of images. The constructing comprises: obtaining a plurality of signal values from the portion of the sample from the plurality of images; evaluating a threshold for the portion; and evaluating a pixel value by integrating a fraction of the plurality of signal values based on the threshold.