Confocal Microscope Pinhole Disc Path Length Compensation
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Solution Overview
Problem
Conventional spinning disc confocal microscope systems suffer from low light efficiency due to the combination and separation of excitation and emission beams in a finite optical space, which limits their ability to achieve both high light throughput and simple structure.
Innovation Solution
A confocal system design where only emission light passes through pinholes for confocal filtering, while excitation spots are created in a space between discs, using a path length difference compensation arrangement to match the beam paths, allowing for separate and recombined excitation and emission beams to travel on different paths, thus avoiding the need for both beams to pass through the same pinholes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Loss of energy
If both excitation and emission beams pass through the same pinholes, then confocal filtering is achieved, but light efficiency decreases due to the finite optical space required for beam combination and separation
Solution Approach 1:
The patent divides the beam paths by allowing excitation and emission beams to travel on different optical paths. The excitation beam passes through the pinhole disc from one side while the emission beam returns through the same pinholes from the opposite side, effectively segmenting the finite optical space into separate excitation and emission pathways. This resolves the contradiction by maintaining confocal filtering while reducing the required optical space and improving light efficiency.
Solution Approach 2:
The patent utilizes the third dimension (depth along the optical axis) by positioning the pinhole disc such that excitation and emission beams traverse it in opposite directions. This dimensional approach allows both beams to share the same pinholes without requiring additional lateral space for separate beam paths, thereby improving light efficiency while maintaining manageable device complexity.
2Device complexity
If a single disc is used for both micro-lenses and pinholes, then device complexity is reduced, but path length matching between excitation and emission beams becomes difficult
Solution Approach 1:
The patent makes the single disc universal by integrating both micro-lenses and pinholes into one component. The disc performs dual functions: generating the excitation spot pattern through micro-lenses and providing confocal filtering through pinholes. This multi-functionality reduces device complexity while the specific design ensures that path length matching is achieved through the geometric arrangement of these elements on the same disc.
Solution Approach 2:
The patent adjusts the radial positions of micro-lenses and pinholes on the disc, as well as the disc's position relative to the objective, to optimize path length matching. By changing these geometric parameters, the system achieves precise path length equality between excitation and emission beams while maintaining a simple single-disc structure.
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 design enhances light efficiency and maintains a simple structure by ensuring precise path length matching between excitation and emission beams, improving image quality and resolution without the limitations of previous systems.
Implementation Method 1
micro-lenses in a corresponding second disk to concentrate excitation intensity onto the pinholes of the first disk
Implementation Method 2
confocal spatial filtering by pinholes carried by the same disc
Implementation Method 3
both discs rotate around a common axis, one obtains many (typically more than 1000) simultaneously illuminated spots on the sample which—by rotation—sequentially illuminate the entire area of the sample seen by the detector
Data Source
AI summary
A system for confocal observation of a sample, having a mask, which is arranged in both an illumination beam path and in an image beam path and which is rotatable around a central axis, openings arranged in a pinhole plane, an arrangement of focusing micro-optics aligned with the geometric arrangement of the openings of the mask to generate a focal pattern, and an optical assembly with an objective, and which is configured to image the focal pattern onto the sample for generating an illumination pattern moving across the sample and to collect light from the sample by the objective and to image it onto the openings of the mask for confocal filtering. The focal plane and pinhole plane are separated by a distance and the optical assembly has a path length difference compensation arrangement which is selective with regard to the illumination light and the light collected from the sample.