Laser Scanning Microscope Dual Pinhole Confocal Filtration
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Solution Overview
Problem
Conventional confocal laser scanning microscopes face challenges in achieving complete confocality and image quality due to fixed pinhole sizes and distances, limiting variability in scanning areas and multifocal mode operations.
Innovation Solution
The implementation of two optically conjugate pinhole planes with variable apertures and a geometric orientation rotation of image points by 90° using miniaturized image-rotating prisms or periscopic mirror arrangements, allowing for adjustable spot distances and orientations without compromising confocal filtration.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If fixed pinhole sizes are used, then manufacturing is simplified, but adaptability to different scanning areas and multifocal modes is limited
Solution Approach 1:
The patent replaces fixed pinholes with variable aperture elements (such as liquid crystal shutters or micro-electromechanical systems) that can dynamically adjust their size and shape. This allows the system to adapt to different scanning areas and multifocal modes while maintaining manufacturing simplicity, as the aperture elements are integrated into the existing pinhole plane structure.
Solution Approach 2:
The invention introduces the ability to change aperture parameters (size, shape, position) electronically or mechanically after manufacturing. This parameter variability is achieved through controllable aperture elements that can be adjusted via control signals, enabling adaptation to different scanning configurations without requiring different physical pinhole sizes.
2Manufacturing precision
If the distance between pinholes is fixed, then manufacturing precision requirements are reduced, but the scanning area viewed cannot be varied
Solution Approach 1:
The patent implements movable or adjustable aperture elements that can change their positions relative to each other. This dynamic positioning capability allows the system to view different scanning areas by adjusting the effective pinhole array configuration, while the physical pinholes themselves can be manufactured with standard tolerances.
Solution Approach 2:
The invention divides the pinhole array into independently controllable segments or groups that can be selectively activated or positioned. This segmentation allows different subsets of pinholes to be used for different scanning areas, providing variability without requiring precise manufacturing of the entire array for each configuration.
3Reliability
If maximum pinhole size is limited by pinhole distance, then confocal filtration is maintained, but flexibility in aperture selection is reduced
Solution Approach 1:
The patent employs variable aperture elements that can dynamically adjust their size within the constraints of the pinhole spacing. This allows the system to select from a range of effective aperture sizes while maintaining the confocal filtration property, as the maximum aperture size is determined by the physical pinhole distance rather than being fixed at a single value.
4Reliability
If multiple pinhole planes are introduced, then complete confocality in orthogonal directions is achieved, but device complexity increases
Solution Approach 1:
The patent combines multiple pinhole planes into a single integrated structure where the pinholes are arranged in a three-dimensional configuration. This merging reduces the complexity of aligning separate planes while maintaining complete confocal filtration, as the pinholes are positioned relative to each other within a unified optical path.
Solution Approach 2:
The invention extends the pinhole arrangement from a two-dimensional plane to a three-dimensional structure, with pinholes distributed across multiple depths or layers. This dimensional extension enables confocal filtration in orthogonal directions without requiring separate adjustable planes, as the third dimension provides the necessary spatial separation and filtering capability.
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 solution enables complete confocal filtration in orthogonal directions, reduces manufacturing tolerances' impact on image quality, and allows for slight variability in scanning area sizes, enhancing the flexibility and efficiency of multifocal operations.
Implementation Method 1
fluorescing samples in particular are stimulated to emit light
Implementation Method 2
The fluorescence, originating from the sample, is imaged on a confocal aperture
Data Source
AI summary
A laser scanning microscope (LSM) consisting of at least one light source from which an illumination beam path extends in the direction of a sample, at least one detection beam path for transmitting sample light to a detector array, a first pinhole for confocal filtering in front of the detector array, a scanner for causing a relative motion between the illumination light and the sample in at least one direction, and a microscope lens. For illuminating a sample, at least two illumination beams, which the microscope lens focuses as illumination points in a sample plane, are generated in the illumination beam path. The laser scanning microscope is characterized in that in addition to the preferably adjustable, slit-shaped first pinhole, a second, preferably adjustable, slit-shaped pinhole is arranged downstream of the first pinhole so as to create optically conjugate beams arranged between the first and the second pinhole.