Fluorescence Microscopy Without Dichroic Mirrors
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Solution Overview
Problem
Conventional fluorescence microscopic imaging apparatuses have complex structures and high costs due to the need for dichroic mirrors to separate light, leading to light energy loss and less accurate fluorescence images.
Innovation Solution
A fluorescence microscopic imaging method and apparatus that eliminates the use of dichroic mirrors by using multiple monochromatic fluorescence excitation light sources arranged around the central axis of the imaging light path, with the light sources intersecting at a preset position on the sample platform, allowing oblique illumination and reducing the amount of excitation light entering the objective lens, and incorporating an emitting light filtering module to filter and acquire fluorescence images.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If a dichroic mirror is used to separate excitation light from fluorescence, then light separation is achieved, but the structure becomes complicated and cost increases
Solution Approach 1:
The patent removes the dichroic mirror from the optical path entirely. Instead of using a dichroic mirror to separate excitation light from fluorescence, the system uses multiple monochromatic light sources positioned at different angles around the objective lens, allowing each light source to illuminate the sample from a specific direction without requiring light separation components
Solution Approach 2:
The patent divides the illumination function into multiple independent monochromatic light sources, each responsible for a specific excitation wavelength. These light sources are arranged around the objective lens and illuminate the sample from different angles, eliminating the need for a single complex dichroic mirror system
2Reliability
If a dichroic mirror is used to separate light, then excitation light can be reflected to the objective lens, but light energy is lost
Solution Approach 1:
The dichroic mirror is completely removed from the system. The patent uses direct illumination from multiple angles where excitation light travels directly from the light sources through the sample to the objective lens without reflection, eliminating energy loss at the dichroic mirror interface
Solution Approach 2:
Instead of using a single light source with a dichroic mirror to redirect light, the patent inverts the approach by using multiple light sources positioned to illuminate the sample directly from different angles, eliminating the need for light redirection and associated energy losses
3Measurement precision
If monochromatic fluorescence excitation light sources are arranged around the central axis of the imaging light path, then oblique illumination is achieved, but the arrangement complexity increases
Solution Approach 1:
The illumination system is segmented into multiple independent monochromatic light sources positioned at different angles around the objective lens. Each light source illuminates the sample from a specific direction, allowing precise control over the illumination geometry and reducing excitation light interference in the imaging path
Solution Approach 2:
The multiple light sources serve dual functions: they provide oblique illumination to reduce excitation light interference while also allowing selective activation of specific wavelengths. The control system enables flexible configuration of which light sources are active based on the fluorescence markers being detected
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 a simpler, cost-effective apparatus that minimizes light energy loss and enhances the accuracy and brightness of fluorescence images by reducing the influence of excitation light during imaging.
Implementation Method 1
monochromatic fluorescence excitation light sources... monochromatic fluorescence excitation light emitted by each of the multiple monochromatic fluorescence excitation light sources obliquely enters a preset detection region of the to-be-detected sample plate... fluorescence of particles within the preset detection region excited by irradiation of monochromatic fluorescence excitation light
Implementation Method 2
collecting, at a side of the to-be-detected sample plate facing away from the target light source, fluorescence of particles within the preset detection region excited by irradiation of monochromatic fluorescence excitation light emitted by the target light source, and magnifying the preset detection region a preset number of times
Implementation Method 3
filtering the excited fluorescence of the particles within the preset detection region... acquiring a fluorescence image of the preset detection region
Data Source
AI summary
A fluorescence microscopic imaging method includes: after a to-be-detected sample plate is placed, lightening, according to experimental requirements, at least one monochromatic fluorescence excitation light source with a same color among multiple monochromatic fluorescence excitation light sources as a target light source, where monochromatic fluorescence excitation light emitted by each monochromatic fluorescence excitation light source obliquely enters a preset detection region of the to-be-detected sample plate; collecting, at a side of the to-be-detected sample plate facing away from the target light source, fluorescence of particles within the preset detection region excited by irradiation of monochromatic fluorescence excitation light emitted by the target light source, and magnifying the preset detection region a preset number of times; filtering the excited fluorescence of the particles within the preset detection region; and acquiring a fluorescence image of the preset detection region.