Molecularity Measurement Instrument Using Image Detector Calibration
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Solution Overview
Problem
Current micro-array scanners face challenges in accurately measuring and comparing fluorescent light quantities due to the lack of direct correlation between excitation and emission light, and they cannot provide absolute values for molecular counting, limiting the ability to compare measurements across different instruments and over time.
Innovation Solution
A method and instrument for quantitatively measuring molecularity by calculating the theoretical light quantity per molecule using molar absorption coefficients, quantum efficiency, and excitation wavelength efficiency, and measuring the light quantity with an image detector to derive molecularity through a calibrated ratio.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If a micro-array scanner uses a photomultiplier to measure fluorescent light quantity, then the measurement can be performed, but it is difficult to obtain correlation between the fluorescent light quantity and the excitation light quantity irradiated on the sample
Solution Approach 1:
The patent introduces an image detector as an intermediary device that can measure both the fluorescent light quantity emitted by the sample and the excitation light quantity irradiated on the sample. This intermediary measurement capability enables the establishment of correlation between excitation and emission light quantities, resolving the technical contradiction by providing a new measurement approach that maintains precision while managing system complexity through integrated measurement.
Solution Approach 2:
The patent employs an image detector that serves multiple functions: it detects fluorescent light emission from the sample, measures excitation light intensity, and provides spatial information. This multi-functional approach allows the system to obtain correlated measurements without requiring separate dedicated devices for each measurement type, thereby improving measurement precision while controlling device complexity.
2Measurement precision
If a photodiode is used to measure uniform light quantity traceable to national standard, then absolute light quantity values can be obtained, but only point data in zero dimension can be obtained without image information
Solution Approach 1:
The patent merges the capabilities of point-based absolute measurement (through calibration with national standards) and image-based spatial measurement into a single integrated system using an image detector. This combination allows the system to obtain both absolute light quantity values traceable to national standards and spatial distribution information simultaneously, eliminating the loss of spatial information while maintaining measurement precision.
Solution Approach 2:
The patent transitions from zero-dimensional point measurement to two-dimensional image measurement by using an image detector. This dimensional change enables the system to capture spatial distribution information across the sample while maintaining the ability to measure absolute light quantity values through calibration, thus resolving the contradiction between measurement precision and information completeness.
3Adaptability or versatility
If conventional methods are used, then relative measurements within one screen can be taken, but direct comparison between data blocks from respective measurements taken a plurality of times and by use of a plurality of apparatuses is not possible
Solution Approach 1:
The patent changes the measurement parameter from relative units to absolute units traceable to national standards. By calibrating the image detector against national standards, the system establishes a unified measurement scale that enables direct comparison between data blocks from multiple measurements and multiple apparatuses, thereby improving adaptability while maintaining measurement precision through standardized absolute values.
4Loss of information
If a camera is used to capture DNA micro-array images, then spatial distribution information can be obtained, but the light quantity measured lacks direct correlation with excitation light quantity and cannot provide absolute molecular counting
Solution Approach 1:
The patent introduces a feedback mechanism through calibration measurements. By measuring the response of the image detector to known light quantities (through calibration with national standards), the system establishes a conversion factor that enables absolute molecular counting. This feedback from calibration data allows the camera to transform relative pixel values into absolute light quantity values, thereby achieving both spatial distribution information and absolute counting capability.
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 absolute value measurements traceable to national standards, allowing for direct comparison of fluorescent light quantities and molecular counting across different instruments and over time, while also enabling spatial distribution analysis of fluorescence molecularity in two or three dimensions.
Implementation Method 1
measuring a light quantity of the sample by use of an image detector
Implementation Method 2
calculating the theoretical light quantity per molecule using molar absorption coefficients
Implementation Method 3
fluorescent light quantity of fluorescence emitted from the sample
Data Source
AI summary
There is provided a molecularity measurement instrument capable of working out the number of molecules in a sample by comparing a measured value of a light quantity with a theoretical light quantity per a single molecule, and a molecularity measurement method using the same. The molecularity of the sample is quantitatively estimated on the basis of a light quantity having correlation with the molecularity. The molecularity measurement method comprises the step of working out a theoretical light quantity per a single molecule, the step of measuring a light quantity of the sample by use of an image detector, and the step of working out the molecularity of the sample on the basis of a ratio of the light quantity of the sample to the theoretical light quantity as worked out.