Colocalised Fluorescence Signal Visualization via Regression Mapping
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Solution Overview
Problem
Current methods for visualizing colocalized fluorescence signals in fluorescence microscopy lack effective spatial representation and intensity correlation analysis, particularly in three-dimensional spaces, leading to ambiguity and variability in data interpretation.
Innovation Solution
A computer-implemented method that calculates a regression parameter to quantify the correlation between fluorescence channel intensities, assigns colormap values based on this correlation, and renders voxels in a 3D image to visualize colocalization, excluding background noise and outliers, and emphasizing positively correlated intensities.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If simple overlay of fluorescence channels is used for visualization, then rapid overview of colocalised signals is achieved, but the ability to observe overlap is highly dependent on relative signal intensity making interpretation ambiguous
Solution Approach 1:
The patent applies color mapping transformations to fluorescence channel data, where each voxel is assigned a color based on its intensity values in multiple channels. The color mapping function transforms the raw fluorescence intensities into a visual representation where color hue, saturation, and brightness encode different aspects of colocalisation, allowing accurate assessment independent of absolute signal intensities.
Solution Approach 2:
The patent introduces an intermediary color mapping layer between the raw fluorescence data and human perception. This color mapping acts as a mediator that translates complex multi-channel intensity relationships into intuitive visual cues, enabling researchers to accurately assess colocalisation without being confounded by variations in absolute signal intensity.
2Measurement precision
If binary mask visualization is used to show colocalisation location, then spatial position is clearly indicated, but no indication is provided of underlying intensities or extent of correlation
Solution Approach 1:
The patent applies local quality by assigning different color attributes to different aspects of coloxicalisation at each voxel location. Specifically, the hue of the color indicates the presence of coloxicalisation, while saturation and brightness encode the intensity values and correlation extent, allowing multiple pieces of information to be displayed simultaneously at each spatial location.
Solution Approach 2:
The patent uses asymmetric color encoding where different color properties (hue, saturation, brightness) represent different types of information. This asymmetric mapping allows the visualization to convey both spatial location and intensity/correlation information in a single integrated display, rather than treating all information symmetrically.
3Device complexity
If traditional 2D visualization is used for colocalisation, then simple display is achieved, but limited spatial information is provided compared to 3D space
Solution Approach 1:
The patent extends the visualization from 2D to 3D by processing and displaying coloxicalisation data in three-dimensional space. Each voxel in the 3D volume is assigned a color based on its intensity values, allowing researchers to visualise coloxicalisation patterns throughout the entire volume rather than just on a projected plane, preserving all spatial information.
4Measurement precision
If coloxicalisation metrics are calculated to quantify coloxicalisation degree, then accurate quantification is achieved, but spatial information is not conveyed
Solution Approach 1:
The patent segments the coloxicalisation analysis into voxel-level assessments, where each voxel is independently evaluated for its coloxicalisation properties and assigned a corresponding color. This segmentation allows the visualization to convey both the degree of coloxicalisation (through color properties) and the spatial distribution (through voxel positions) simultaneously.
Data Source
AI summary
A computer-implemented method and a system are provided for visualising colocalised fluorescence signals. The method accesses signal intensity data obtained from a first fluorescence channel and a second fluorescence channel in which the signal intensity data is associated with voxels in an image. A regression factor on the signal intensity data is calculated to generate a regression parameter corresponding to a degree of correlation between the signal intensity data obtained from the first and second fluorescence channels The signal intensity data is mapped to the regression parameter and colourmap values are assigned to each voxel based on the mapped signal intensity data in which colourmap values of voxels embodying poorly correlated signal intensity data are reduced. The method renders the voxels in the image in colours according to their colourmap values to visualise colocalisation in the image.