Confocal Microscope Autofluorescence and Reflected Light Data Association
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Solution Overview
Problem
Existing methods for identifying and analyzing microorganisms lack the ability to non-invasively acquire spatial positional information in three-dimensional space, and do not effectively record autofluorescence and reflected light in association with each other on a single set of coordinates.
Innovation Solution
A data creation method that uses a confocal laser scanning microscope to irradiate a specimen with laser light, obtaining autofluorescence and reflected light, and generating data that associates the intensity of this light with spatial coordinates, allowing for non-invasive analysis and identification of microorganisms.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional metagenomic analysis or colony autofluorescence methods are used to identify microorganisms, then species identification can be achieved, but spatial positional information in three-dimensional space cannot be acquired
Solution Approach 1:
The patent combines multiple types of light detection (autofluorescence and reflected light) into a single integrated system that captures both signals simultaneously with their spatial coordinates, merging previously separate identification and imaging functions into one comprehensive measurement approach
Solution Approach 2:
The invention transitions from conventional two-dimensional imaging to three-dimensional spatial mapping by utilizing the confocal microscope's depth scanning capability to acquire autofluorescence and reflected light signals at multiple focal planes, thereby reconstructing three-dimensional positional information
2Adaptability or versatility
If autofluorescence and reflected light are recorded separately, then each signal can be analyzed independently, but they cannot be associated with each other on one set of coordinates
Solution Approach 1:
The system uses the detected reflected light signal as feedback to identify and extract corresponding autofluorescence signals from the same spatial location, creating a coordinated relationship between the two signal types through iterative detection and matching processes
Solution Approach 2:
The patent creates a universal data structure that can accommodate both autofluorescence and reflected light signals within the same coordinate system, enabling the system to handle multiple signal types simultaneously while maintaining their spatial relationships for versatile analysis
3Measurement precision
If confocal microscope is used to observe microorganism dynamics in three-dimensional space, then positional and movement information can be grasped, but non-invasive identification based on autofluorescence at single-cell resolution has not been reported
Solution Approach 1:
The system performs preliminary scanning to detect reflected light signals that indicate the presence and position of microorganisms, then uses this preliminary information to guide subsequent autofluorescence detection at the same locations, ensuring accurate single-cell identification
Solution Approach 2:
The reflected light signal serves as an intermediary that bridges the gap between physical presence detection and biochemical identification, allowing the system to locate microorganisms and then identify them through their autofluorescence characteristics without direct contact or intervention
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 non-invasive analysis and identification of microorganisms by acquiring spatial positional information in three-dimensional space, effectively utilizing autofluorescence and reflected light data to distinguish between different species of microorganisms.
Implementation Method 1
irradiating a specimen with laser light, obtaining autofluorescence and reflected light
Implementation Method 2
irradiating a specimen with laser light, obtaining autofluorescence and reflected light
Data Source
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AI summary
A data creation method includes: an autofluorescence data generation step of placing a focus of light having a predetermined wavelength at one set of coordinates on a predetermined focal plane, irradiating a sample positioned at the set of coordinates with excitation light containing the light to obtain autofluorescence emitted from the sample, and generating autofluorescence data including intensity data and/or spectrum data of the autofluorescence; a reflected light data generation step of irradiating the set of coordinates on the predetermined focal plane with illumination light to obtain reflected light scattered by the sample, and generating intensity data of the reflected light; and a correspondence data creation step of creating correspondence data associating the autofluorescence data and the reflected light data on the set of coordinates on the predetermined focal plane.