STORM Fluorescent Image Analysis Using Bright Spot Area Quantification
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Solution Overview
Problem
Conventional STORM microscopy methods cannot perform quantitative analysis on high-resolution fluorescent images due to bright spot data being localized to particular coordinates, preventing basic arithmetic operations using pixel intensity values.
Innovation Solution
An analysis device and method that sets predetermined areas corresponding to the positions of bright spots in STORM images, allowing for quantitative analysis by associating position information with intensity values and performing calculations on these areas.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If STORM microscopy is used to achieve high resolution, then spatial resolution is improved, but quantitative analysis capability deteriorates because bright spot data is localized to coordinates without pixel intensity values
Solution Approach 1:
The patent segments the high-resolution STORM image into multiple low-resolution images corresponding to different time points. Each low-resolution image contains pixel intensity values that can be used for quantitative analysis. This segmentation allows the system to maintain high spatial resolution through STORM while recovering quantitative information by distributing bright spot data across multiple temporal segments with associated intensity values.
Solution Approach 2:
The patent introduces a temporal dimension to resolve the contradiction. By distributing bright spot detections across multiple time points and associating each with pixel intensity values from corresponding low-resolution images, the system transforms the problem from a purely spatial coordinate system to a spatio-temporal system. This allows quantitative analysis to be performed on intensity values while maintaining high spatial resolution through the STORM reconstruction.
2Measurement precision
If bright spot data is localized to coordinates for high resolution imaging, then spatial precision is improved, but arithmetic operations on pixel intensity values become impossible
Solution Approach 1:
The patent performs preliminary action by capturing multiple low-resolution fluorescent images at different time points before reconstructing the high-resolution STORM image. These preliminary low-resolution images retain pixel intensity information that is subsequently associated with bright spot positions in the high-resolution reconstruction. This preliminary capture of intensity data prevents information loss while maintaining position accuracy.
Solution Approach 2:
The patent uses low-resolution fluorescent images as intermediaries to bridge the gap between high-resolution position data and quantitative intensity analysis. These intermediary images serve as a bridge, containing pixel intensity values that can be mathematically associated with bright spot coordinates in the STORM reconstruction, thereby preserving both position accuracy and intensity information.
3Measurement precision
If multiple fluorescent images are acquired repeatedly for STORM reconstruction, then high resolution is achieved, but time consumption increases
Solution Approach 1:
The patent applies partial action by acquiring fluorescent images with lower temporal resolution than would be required for complete STORM reconstruction. Instead of capturing every possible frame needed for maximum resolution, the system captures a subset of images at strategically selected time points. This partial acquisition is sufficient to provide the temporal distribution of bright spots needed for quantitative analysis while reducing overall acquisition time.
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 quantitative analysis of high-resolution STORM images by defining spatial and temporal areas around bright spots, facilitating accurate comparisons and evaluations.
Implementation Method 1
A fluorescent material or object with said fluorescent material adhered thereto is used as an observation sample. This fluorescent material has properties whereby it becomes active when irradiated with activation light of a predetermined wavelength and, thereafter, becomes inactive by emitting fluorescence when irradiated with excitation light of a wavelength different from the activation light.
Data Source
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AI summary
An analysis device for quantifying a state of a fluorescent image containing a plurality of bright spots comprises an area setting unit in which states of a plurality of bright spots contained in a plurality of areas set in the fluorescent image in accordance with positions of the plurality of bright spots are quantified as numerical values.