Fluorescence Microscopy Scanning System with Separate Optics
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Solution Overview
Problem
Conventional fluorescence microscopy systems face challenges in efficiently analyzing large numbers of samples while discriminating against background noise, particularly in industrial settings where the existing configurations are not suitable or cost-effective.
Innovation Solution
A scanning light source system with improved optical geometry and separate illumination and collection optical components, allowing for efficient scanning and detection of fluorescence signals from specific vertical depths within a sample, using a combination of pulsed or modulated lasers and advanced optics like cylindrical lenses and optical fibers to enhance spatial resolution and reduce background interference.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional fluorescence microscopy systems are used, then fluorescence signals can be detected, but background noise cannot be effectively discriminated against and analysis of large numbers of samples is inefficient
Solution Approach 1:
The patent segments the detection process by using multiple detectors positioned at different vertical depths to collect fluorescence signals from different focal regions simultaneously. This segmentation allows specific signal discrimination from background noise while enabling parallel processing of multiple sample locations, thus improving both measurement precision and productivity
Solution Approach 2:
The patent introduces a vertical depth dimension to the detection system by positioning detectors at different heights above the sample surface. This dimensional addition enables selective collection of fluorescence signals from specific focal planes, improving signal discrimination while allowing rapid scanning across multiple sample positions
2Measurement precision
If confocal laser scanning microscopy or wide-field deconvolution technologies are used to improve resolution and eliminate background noise, then optical slices can be generated, but the systems are complex and not suitable for routine imaging or industrial applications
Solution Approach 1:
The patent extracts only the essential function of optical sectioning by using simple aperture-based detection at specific vertical depths, eliminating the need for complex confocal pinholes or computational deconvolution algorithms. This extraction maintains spatial resolution capability while dramatically reducing system complexity for routine and industrial applications
Solution Approach 2:
The patent replaces expensive, complex optical systems with simpler, more cost-effective components including basic apertures, standard detectors, and straightforward scanning mechanisms, making the system suitable for routine imaging and industrial deployment
3Volume of moving object
If two-photon excitation with infrared ultra-short pulsed laser is used, then penetration depth is improved and photo bleaching is reduced, but the system complexity increases and cost efficiency decreases for routine imaging
Solution Approach 1:
The patent achieves effective deep-tissue imaging by using visible light excitation with carefully controlled optical sectioning that copies the beneficial effects of two-photon microscopy (reduced scattering, improved penetration) through simpler means, eliminating the need for expensive ultra-short pulsed infrared lasers while maintaining comparable performance for routine applications
4Device complexity
If illumination light and fluorescent light share an optical path, then the system is simpler, but background noise from excitation light cannot be effectively eliminated
Solution Approach 1:
The patent employs dynamic temporal separation by using pulsed or modulated excitation light sources synchronized with gated detection windows. This dynamic approach allows the system to maintain a shared optical path while effectively discriminating fluorescent signals from excitation light background through time-resolved detection
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
Enables efficient analysis of multiple samples with improved spatial resolution and reduced background noise, allowing for precise mapping of fluorescence characteristics and rapid assay analysis without the need for conventional autofocus mechanisms, suitable for both cellular and microarray analysis.
Implementation Method 1
The illumination light, which typically is provided from a laser, excites the fluorophores into a higher energy state where they remain for a short period of time, before returning to their original energy state while emitting fluorescent light of a wavelength longer than the excitation wavelength.
Implementation Method 2
The systems that have found most use in laboratories generally use visible fluorescence of materials and visible light sources. The illumination light, which typically is provided from a laser, excites the fluorophores
Implementation Method 3
both the excitation light and the fluorescent light share an optical path through the microscope's optical system, and can be separated as needed, by optical components such as dichroic mirrors that reflect light above the excitation wavelengths while passing the excitation light
Implementation Method 4
the infrared wavelength excitation significantly reduces scattering within the tissue as the scattering coefficient is proportional to the inverse fourth power of the excitation wavelength, resulting in penetration deeper into the specimen
Data Source
AI summary
Methods and apparatus, including computer program products, implementing and using techniques for collecting optical data pertaining to one or more characteristics of a sample. A light beam of a first frequency is scanned onto a sample surface using one or more illumination optical elements. Light of a second frequency is collected from a scan line on the sample surface using one or more collection optical elements. None of the one or more collection optical elements are included among the one or more illumination optical elements. The collected light is transmitted to a detector.