Fluorescence Reading Device Height Difference Detection
Solution Overview
Problem
Existing microarray analysis methods struggle with accurate alignment, especially when the sample contains a small amount of DNA or when the DNA chip lacks a positive control, leading to incomplete or inaccurate data acquisition due to insufficient fluorescence intensity from DNA probes.
Innovation Solution
A fluorescence reading device and method that acquires alignment image data by positioning the DNA chip closer to the objective lens than the focal position, allowing for the detection of height differences on the substrate surface, enabling accurate alignment and analysis even with low DNA amounts or without positive controls.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Illumination intensity
If the DNA chip is positioned at the focal position of the objective lens, then the fluorescence intensity is maximized, but the height difference detection accuracy deteriorates due to insufficient depth of field
Solution Approach 1:
The patent transitions from a single focal plane to multiple focal planes by positioning the DNA chip at different distances from the objective lens. By capturing images at multiple depths (including out-of-focus images), the system enables both fluorescence intensity measurement and height difference detection through optical sectioning and image processing techniques.
2Measurement precision
If the DNA chip is positioned closer to the objective lens than the focal position, then the height difference detection accuracy is improved, but the fluorescence intensity decreases
Solution Approach 1:
The patent performs preliminary acquisition of multiple images at different focal positions before final analysis. By capturing a series of images at varying depths (including positions before and after the focal point), the system prepares the necessary data to subsequently extract both height information and fluorescence intensity through image processing, resolving the intensity loss at non-focal positions.
3Measurement precision
If multiple images at different focal positions are acquired, then the height difference and fluorescence intensity can be accurately measured, but the measurement time increases
Solution Approach 1:
The patent employs periodic scanning of the DNA chip through the focal point by moving the chip in regular intervals at different speeds. This systematic periodic movement allows efficient acquisition of multiple focal planes in a structured manner, reducing redundant measurements and optimizing the time required to capture sufficient depth information for accurate height and intensity analysis.
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 ensures proper alignment and analysis of DNA chips with small DNA amounts or without positive controls, enhancing data accuracy and reliability by clearly detecting edge shapes and correcting image strain.
Implementation Method 1
an objective lens 13 for focusing light
Implementation Method 2
An optically detectable luminescent or fluorescent mark is added to the DNA sample
Data Source
Figure 1
Figure 2(a)~2(c)
Figure 3
AI summary
A detecting method, a microarray analyzing method, and a fluorescence reading device, which enable acquirement of an image, from which a height difference of a substrate is detectable, are provided. In a detecting method of irradiating a substrate having a concave and convex shape with laser light collected by a lens, acquiring, as image data, received light intensity of reflected light and/or scattered light from the substrate, and detecting a height difference of the concave and convex shape, a light irradiation surface of the substrate is arranged at a position closer to the lens than a focal position of the lens is, reflected light and/or scattered light from the light irradiation surface is received as detected light, and a height difference of the substrate is detected based on a change in intensity of the received light.