Fluorescence Normalization in Nucleic Acid Sequence Measurement
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Solution Overview
Problem
Nucleic acid sequence measurement devices experience variations and changes in spot light intensity due to manufacturing inconsistencies and environmental factors, leading to decreased detection sensitivity and reliability through false positives or negatives.
Innovation Solution
Incorporating a measurement area that emits fluorescence upon target reaction and a reference area that emits fluorescence independently of target reaction, with a calculator normalizing and correcting light intensity using the reference area's fluorescence to stabilize measurements.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If a nucleic acid sequence measurement device is used to measure target sequences, then detection capability is provided, but variations in spot light intensity occur due to manufacturing inconsistencies and environmental factors
Solution Approach 1:
A reference area containing reference probes is introduced as an intermediary element. This reference area emits fluorescence independently of target hybridization and serves as a mediator to detect and compensate for variations in spot light intensity. By comparing the fluorescence intensity in the measurement area with that in the reference area, the system can normalize the data and eliminate false positives or negatives caused by light intensity variations.
2Measurement precision
If detection probes are provided on a solid phase surface for hybridization-based measurement, then target detection capability is achieved, but false positives or negatives occur due to light intensity variations
Solution Approach 1:
The reference area provides continuous feedback information about the actual light intensity conditions in each spot. This feedback mechanism allows the system to dynamically adjust and normalize the measurement signals, enabling accurate target detection even when light intensity varies across different spots or over time.
3Ease of manufacture
If spot light intensity varies between manufacturing lots and devices, then manufacturing flexibility is maintained, but measurement consistency deteriorates
Solution Approach 1:
Instead of attempting to control and maintain absolutely uniform spot light intensity during manufacturing, the invention changes the approach by introducing a reference area that captures the actual light intensity parameter. This allows the system to adapt to manufacturing variations without requiring tight process control, thereby maintaining manufacturing flexibility while achieving measurement consistency through software-based normalization.
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
Ensures reliable nucleic acid sequence measurement by compensating for variations and changes in spot light intensity, enhancing detection accuracy and reducing false results.
Implementation Method 1
a detection probe having a complementary sequence to the specific nucleic acid sequence described above is provided on a solid phase surface such as a substrate. In this method, a sample containing a target is added to a DNA chip, and the target is measured by using the properties of the target reacted by the detection probe of the DNA chip due to hybridization.
Implementation Method 2
a detector (12) configured to detect fluorescence emitted from a nucleic acid sequence measurement device (DV)
Data Source
Figure 1
Figure 2~3
Figure 4~5(b)
AI summary
A nucleic acid sequence measurement apparatus that measures a target with a specific nucleic acid sequence contained in a sample. The nucleic acid sequence measurement apparatus includes: a detector configured to detect fluorescence emitted from a nucleic acid sequence measurement device, which is provided with a measurement area that emits first fluorescence due to a reaction with the target, and a reference area that emits second fluorescence regardless of the reaction with the target; and a calculator configured to measure the target on the basis of a light intensity obtained by correcting or normalizing a light intensity of the first fluorescence emitted from the measurement area using a light intensity of the second fluorescence emitted from the reference area in the fluorescence detected by the detector.