Spinning Disk Confocal Microscopy Illumination Uniformity
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Solution Overview
Problem
Spinning disk confocal microscopy systems face limitations in illumination intensity and speed, leading to reduced fluorescence and slower imaging of live samples due to the spreading of excitation light over multiple spots, which results in less intense illumination and slower imaging.
Innovation Solution
The use of a liquid light guide and a reflecting mirror tube to direct light from a light source to the confocal optics, providing more uniform illumination by altering the optical path and reducing spatial and temporal coherence, and incorporating a light engine with multiple adjustable light sources and microlenses to enhance brightness and image acquisition.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If spinning disk confocal microscopy uses multiple spots and pinholes for simultaneous imaging, then imaging speed is improved, but illumination intensity is reduced
Solution Approach 1:
The system divides the illumination into multiple discrete spots arranged in a pattern on the spinning disk, allowing simultaneous illumination of multiple regions. This segmentation enables parallel imaging across multiple spots while maintaining focused illumination at each spot through the confocal pinhole arrangement.
Solution Approach 2:
The patent introduces a spatial dimension by arranging multiple spots in a two-dimensional array on the spinning disk. This allows the system to illuminate multiple locations simultaneously without increasing the intensity at each individual spot, as the light is distributed across multiple spatial positions rather than concentrated at a single point.
2Speed
If laser light is used for excitation, then imaging speed is improved through rapid scanning, but spatial and temporal coherence causes non-uniform illumination
Solution Approach 1:
The patent converts the harmful effect of laser coherence into a beneficial one by using the coherent light to create sharp, well-defined spots through the confocal pinhole system. The coherence that would normally cause non-uniformity is instead harnessed to produce high-contrast, uniformly illuminated spots through precise optical focusing and pinhole filtering.
Solution Approach 2:
The confocal pinhole acts as an intermediary element between the laser source and the sample. It filters and shapes the coherent laser light to create uniform illumination spots, mediating between the high-coherence laser and the requirement for uniform illumination across the sample surface.
3Area of stationary object
If excitation light is spread over many spots, then imaging coverage is improved, but fluorescence signal is reduced
Solution Approach 1:
The imaging coverage is segmented into multiple discrete spots arranged in a pattern, allowing the system to cover a larger area simultaneously. Each spot maintains focused illumination through the confocal pinhole, ensuring that fluorescence signal intensity is preserved at each location while the overall coverage area is expanded through the multi-spot arrangement.
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 results in improved uniformity of illumination, increased image brightness, and faster image acquisition rates, enabling the capture of high-quality images with reduced blur and enhanced imaging of live samples.
Implementation Method 1
thermal motion of the liquid in the LLG may alter the optical path and scatter light, reducing or eliminating spatial and temporal coherence introduced by the light source
Implementation Method 2
A reflecting mirror tube may similarly have advantages
Data Source
AI summary
A spinning-disk confocal microscopy system, and components thereof, with improved illumination. The system may include a liquid light guide (LLG), a reflecting mirror tube, and/or other light guide directing light from a light source to the system's confocal optics. An LLG may provide certain advantages over other conveyance mechanisms. For example, thermal motion of the liquid in the LLG may alter the optical path and scatter light, reducing or eliminating spatial and temporal coherence introduced by the light source. This, in turn, may create more uniform illumination on samples. A reflecting mirror tube may similarly have advantages.