FLIM Metabolic Imaging of Lesions via Phasor Analysis

Resolve Bottlenecks,
Find Innovative Solutions
Generate Solutions

Solution Overview

Problem

Current methods for measuring the metabolic status of lesions, such as wounds and tumors, lack sufficient spatial resolution and accuracy, particularly in deeper tissue layers, and fail to effectively quantify anaerobic glycolysis markers like the NADH/NAD+ ratio, which is crucial for understanding healing processes and cancer progression.

Innovation Solution

A fluorescence lifetime imaging microscopy (FLIM) based method that acquires and analyzes FLIM data of anaerobic glycolysis markers, using phasor analysis to generate quantitative 3D images of metabolic status, optionally combining with pH, oxygen tension, glucose, pyruvate, and lactate markers, to provide detailed metabolic maps of lesions.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If conventional fluorescence imaging methods are used to measure metabolic status of lesions, then the measurement process is simple, but the spatial resolution and accuracy are insufficient particularly in deeper tissue layers

Engineering Contradiction:
Improvespatial resolution and accuracy of metabolic status measurementVSAvoidcomplexity of FLIM acquisition and phasor analysis system
Core Design Contradiction:
Measurement precisionVSDevice complexity

Solution Approach 1:

The patent replaces conventional intensity-based fluorescence imaging with fluorescence lifetime imaging microscopy (FLIM), which measures the temporal decay of fluorescence rather than spatial distribution. This substitution of measurement principle enables quantitative metabolic status assessment with high precision in deep tissue layers by measuring lifetime parameters that are independent of probe concentration and excitation intensity variations.

Inventive Principle:
Principle #28Mechanics substitution (Replace mechanical system)

Solution Approach 2:

The patent transforms the measurement parameter from fluorescence intensity to fluorescence lifetime. By measuring the temporal decay characteristics of fluorescence emission, the system achieves accurate quantification of metabolic markers (NADH/NAD+ ratio, pH, oxygen tension) without being affected by variations in probe concentration, excitation power, or tissue scattering, thereby resolving the contradiction between measurement precision and device complexity.

Inventive Principle:
Principle #35Parameter changes

2Loss of information

If FLIM data acquisition is performed to obtain metabolic information, then quantitative 3D imaging of metabolic status is achieved, but the data processing and analysis complexity increases

Engineering Contradiction:
Improvecompleteness of metabolic status informationVSAvoidcomplexity of data processing and analysis
Core Design Contradiction:
Loss of informationVSDevice complexity

Solution Approach 1:

The patent introduces phasor analysis as an intermediary computational method that transforms complex FLIM decay curves into simplified 2D phasor plots. Each pixel's lifetime distribution is represented as a point in the phasor diagram, enabling intuitive visualization and quantification of metabolic status without requiring complex curve fitting or mathematical transformations. This intermediary approach preserves complete metabolic information while dramatically simplifying data processing.

Inventive Principle:
Principle #24Intermediary (Mediator)

Solution Approach 2:

The patent transforms the temporal dimension of fluorescence decay (time-domain data) into a spatial representation in the phasor domain (complex plane). By mapping lifetime information onto the real and imaginary components of the Fourier transform at a specific frequency, the system converts complex temporal decay profiles into 2D coordinates, enabling efficient processing and visualization of 3D metabolic status information.

Inventive Principle:
Principle #17Another dimension (Dimensionality change)

3Loss of information

If multiple metabolic markers are combined for comprehensive analysis, then the understanding of healing processes and cancer progression is enhanced, but the measurement and analysis time increases

Engineering Contradiction:
Improvecompleteness of metabolic status informationVSAvoidtime for data acquisition and processing
Core Design Contradiction:
Loss of informationVSLoss of time

Solution Approach 1:

The patent combines multiple metabolic markers (NADH/NAD+ ratio, pH, oxygen tension, glucose, pyruvate, lactate) into a single integrated FLIM measurement process. By using ratiometric probes that report multiple parameters simultaneously and processing all data through unified phasor analysis, the system achieves comprehensive metabolic characterization without requiring separate measurements for each marker, thereby reducing total acquisition and processing time.

Inventive Principle:
Principle #5Merging (Combining)

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 precise, non-invasive, three-dimensional imaging of metabolic activity in wounds and tumors, enhancing the understanding of healing processes and cancer progression, and potentially improving diagnostic and therapeutic strategies by providing detailed spatial distribution of glycolytic activity and pH levels.

Implementation Method 1

The method comprises: a) acquiring fluorescence lifetime imaging microscopy (FLIM) data of an anaerobic glycolysis marker of the lesion

Methodology Applied
Scientific EffectFluorescence: Fluorescence

Data Source

PatentEP4132347B1Method for quantitative imaging of the metabolic status of a lesion
Publication Date: 2024.05.01 HUMANITAS MIRASOLE SPA
  • EP4132347B1 patent drawingFigure 1
  • EP4132347B1 patent drawingFigure 2A~2E
  • EP4132347B1 patent drawingFigure 3A~3D

AI summary

This invention pertains to a fluorescence lifetime imaging microscopy (FLIM) - based method for the quantitative three-dimensional (3D) imaging of the metabolic status of a lesion, comprising the acquisition of FLIM data of a least one anaerobic glycolysis marker. This method applies in particular to lesions such as wounds and tumours, and to markers such as the NADH/NAD+ ratio, the NAD(P)H/NAD+ ratio, the NADH/FAD ratio, the NAD(P)H/FAD ratio or the FADH/FAD ratio as a first marker, and their optional combination with a second marker such as pH, oxygen tension, glucose, pyruvate and lactate.