DNA Microarray Fluorescence Imaging With Photobleaching Correction
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Acquiring a fluorescence image enabling accurate detection is difficult in DNA microarray methods due to photobleaching of fluorescent dyes, especially when long measurement times are required, leading to errors in estimating the amount of detection target molecules.
Innovation Solution
The method involves acquiring a first fluorescence image and a second fluorescence image at different time periods, correcting the second fluorescence intensity based on the first fluorescence intensity, and generating an additively corrected fluorescence image by combining these images to compensate for photobleaching effects.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If a long measurement time period is used to improve detection accuracy, then more sufficient light collection is achieved, but photobleaching of fluorescent dye increases causing error in estimation
Solution Approach 1:
The patent applies preliminary action by capturing a reference fluorescence image before the main measurement to establish baseline intensity. This reference image is taken at an earlier time point when photobleaching has not yet significantly occurred, and is then used to correct the main measurement image, effectively compensating for photobleaching effects without requiring extended measurement time.
Solution Approach 2:
The patent implements feedback by using the reference fluorescence image intensity to correct the main measurement image. The system compares the reference image with the main image, calculates the intensity ratio, and applies this ratio as a correction factor to the main measurement data, thereby compensating for photobleaching effects and improving estimation accuracy.
2Measurement precision
If multiple fluorescence images are acquired at different time periods to correct photobleaching, then detection accuracy improves, but measurement time and processing complexity increase
Solution Approach 1:
The patent applies partial action by acquiring only two fluorescence images (one reference image and one main measurement image) rather than continuous monitoring. This minimal set of images is sufficient to calculate the photobleaching correction factor, balancing measurement accuracy with time efficiency without requiring excessive imaging.
3Reliability
If multiple fluorescence images are acquired and processed to generate corrected images, then photobleaching correction is achieved, but device complexity and processing steps increase
Solution Approach 1:
The patent replaces complex mechanical or chemical photobleaching prevention mechanisms with a computational image processing approach. Instead of using specialized equipment or chemicals to prevent photobleaching, the system uses software algorithms to calculate correction factors from reference images and apply mathematical corrections to main measurement images, simplifying the overall system design.
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 reduces quantitative errors caused by photobleaching, improves detection accuracy, shortens measurement time, and enables comparison of results with different exposure durations.
Implementation Method 1
a first fluorescence image of a measurement target specimen imaged in a first time period by an imaging apparatus
Implementation Method 2
imaging apparatus that acquires a first fluorescence image
Data Source
Figure 1(1)~1(4)
Figure 2(1)~2(2)
Figure 3
AI summary
An image acquisition apparatus 100 acquires a first fluorescence image of a DNA microarray A imaged in a first time period by an imaging apparatus 1 and a second fluorescence image of the DNA microarray A imaged in a second time period by the imaging apparatus, generates a corrected second fluorescence image resulting from correction of a second fluorescence intensity of a fluorescent spot that generates fluorescence in the second fluorescence image, the correction being based on a first fluorescence intensity of a fluorescent spot that generates fluorescence in the first fluorescence image, and generates an additively corrected fluorescence image resulting from correction of the first fluorescence image, by adding the corrected second fluorescence image to the first fluorescence image.