Dichroic Mirror Incident Angle Optimization for Fluorescence Microscopes
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Solution Overview
Problem
High-functional dichroic mirrors used in confocal laser scanning fluorescence microscopes face challenges in separating multiple excitation lights and fluorescence types efficiently, leading to potential distortion of the laser spot, reduced spatial resolution, and decreased detection sensitivity due to the strong control of dielectric multilayer characteristics.
Innovation Solution
A laser excitation fluorescent microscope design with a high-functional dichroic mirror that sets the incident angle of excitation lights and fluorescence between 10° and 25°, preferably 12°, and forms a dielectric multilayer with specific transmittance and reflectance characteristics to optimize separation and reduce film thickness, maintaining spatial resolution and detection sensitivity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If the dielectric multilayer characteristics are strongly controlled to improve separation efficiency, then the separation of excitation lights and fluorescence is improved, but the total film thickness increases causing glass substrate deformation and laser spot distortion
Solution Approach 1:
The patent changes the incident angle parameter from the conventional 45° to a smaller angle (10°-25°, preferably 12°). This parameter change allows the dielectric multilayer to achieve the required separation efficiency with a reduced total film thickness, preventing substrate deformation and laser spot distortion while maintaining high separation performance
Solution Approach 2:
The patent optimizes the dynamic interaction between light and the dielectric multilayer by adjusting the incident angle, which changes how light interacts with each layer. This dynamic adjustment allows for thinner film designs that still achieve the necessary optical separation, reducing the cumulative stress on the substrate
2Manufacturing precision
If the dielectric multilayer characteristics are strongly controlled to improve separation efficiency, then the separation of excitation lights and fluorescence is improved, but the spatial resolution is lowered
Solution Approach 1:
By changing the incident angle parameter to a smaller value (10°-25°), the patent achieves high separation efficiency with thinner film layers, thereby maintaining the laser spot shape and spatial resolution that would otherwise be degraded by excessive film thickness
3Manufacturing precision
If the dielectric multilayer characteristics are strongly controlled, then the reflectivity in reflecting band and transmittance in transmitting band are improved, but the total film thickness increases causing glass substrate deformation
Solution Approach 1:
The patent applies a parameter change to the incident angle (reducing from 45° to 10°-25°), which enables the dielectric multilayer to achieve high reflectivity and transmittance with a reduced total film thickness, thereby minimizing the stress and deformation of the glass substrate
4Manufacturing precision
If the wavelength characteristic curve has ripple to improve separation, then the separation of multiple fluorescence types is improved, but the detection sensitivity is lowered
Solution Approach 1:
By optimizing the incident angle parameter to a smaller value (10°-25°), the patent reduces the ripple in the wavelength characteristic curve while maintaining precise separation wavelengths. This results in both accurate separation of multiple fluorescence types and high detection sensitivity, eliminating the trade-off between separation precision and sensitivity
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
The optimized design enhances the separation efficiency of excitation lights and fluorescence, maintains high spatial resolution, and increases detection sensitivity by reducing film stress and thickness, while also reducing manufacturing costs.
Implementation Method 1
a dielectric multilayer with high efficiency, it is only required to devise to improve a reflectivity in a reflecting band and a transmittance in a transmitting band
Implementation Method 2
a dichroic mirror for separating the excitation lights and a plurality of types of fluorescence generated in accordance with the excitation lights
Implementation Method 3
the objective lens 26 forms a laser spot on the sample S
Implementation Method 4
a sample S on which multistaining procedure is performed using a plurality of types of fluorescent dyes
Data Source
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AI summary
The present invention has a proposition to provide a highly efficient laser excitation fluorescent microscope. Accordingly, a laser excitation fluorescent microscope of the present invention includes a laser light source part (10) radiating at least two types of excitation lights having different wavelengths; a light collecting part (110) collecting the two types of excitation lights radiated by the laser light source part on a sample; a high-functional dichroic mirror (22), disposed between the laser light source part and the light collecting part, reflecting the two types of excitation lights radiated by the laser light source part to make the excitation lights incident on the light collecting part, and transmitting two types of fluorescence generated at the sample in accordance with the two types of excitation lights; and a detecting part (50) detecting light transmitted through the high-functional dichroic mirror, in which an incident angle θ of the excitation lights and the fluorescence to the high-functional dichroic mirror (22) satisfies a formula of 0°<θ<45°.