Microscopy System for Protoporphyrin IX Quantification
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Solution Overview
Problem
Conventional microscopy systems are unable to satisfactorily quantify the fluorescence of protoporphyrin IX and its concentration in tissues, which is crucial for determining the intensity and accumulation of the fluorescent dye in medical applications.
Innovation Solution
A microscopy method and system that utilize two detector fields with different pixel configurations and beam paths, employing a beam splitter and wavelength-dependent filters to separate and quantify the fluorescence of protoporphyrin IX from autofluorescence, using a formula that accounts for radiation intensity and detection efficiencies to determine spatially dependent fluorescence intensity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional microscopy systems are used to detect fluorescence, then the system structure is simple, but the measurement precision of fluorescence quantification is insufficient
Solution Approach 1:
The patent divides the detection system into two separate detector fields with different pixel configurations. The first detector field uses pixels optimized for one wavelength range while the second detector field uses pixels optimized for another wavelength range. This segmentation allows each detector to be specialized for specific spectral detection, thereby improving measurement precision without requiring a completely complex new system architecture.
Solution Approach 2:
Different regions of the detector field are assigned different pixel types with specific wavelength sensitivities. By optimizing local pixel characteristics for specific wavelength ranges, the system achieves superior fluorescence quantification precision in targeted spectral regions while maintaining overall system manageability.
2Adaptability or versatility
If multiple fluorescences are detected simultaneously, then the detection capability is enhanced, but the difficulty of detecting and measuring individual fluorescence intensities increases
Solution Approach 1:
The detection system is segmented into multiple detector fields, each with pixels having different wavelength-dependent detection efficiencies. This allows simultaneous detection of multiple fluorescences with different emission spectra while maintaining the ability to measure individual fluorescence intensities by utilizing the differential response of each pixel type to specific wavelengths.
Solution Approach 2:
The system exploits changes in the detection efficiency parameter across different pixel types and wavelength ranges. By analyzing the pattern of signal intensities across pixels with known different spectral responses, the system can mathematically resolve and quantify individual fluorescence contributions from multiple simultaneously detected fluorescences.
3Measurement precision
If wavelength-dependent filters are introduced to separate fluorescences, then the measurement precision improves, but the device complexity increases
Solution Approach 1:
The patent introduces wavelength-dependent detection efficiency as an intermediary parameter that naturally exists in the detector pixels themselves. Rather than adding complex external filter systems, the system utilizes the inherent spectral selectivity of different pixel types as a mediator to separate and quantify overlapping fluorescences, thereby improving measurement precision without proportionally increasing device complexity.
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 accurate quantification of protoporphyrin IX fluorescence intensity by separating it from autofluorescence, providing precise concentration measurements through the use of distinct wavelength-dependent detection efficiencies and filter characteristics.
Implementation Method 1
the beam splitter may have dichroic properties such that the beam splitter has different wavelength-dependent transmission characteristics for the first beam path and the second beam path
Implementation Method 2
At least one first optical filter with a wavelength-dependent transmission characteristic is arranged in the first beam path between the object region and the first detector field
Implementation Method 3
one value is determined in each case for a plurality of pixels or a plurality of groups of pixels in the first detector field, the value representing a fluorescence intensity
Implementation Method 4
exciting at least two fluorescences in the object region, wherein a first of the two fluorescences may be the fluorescence of protoporphyrin IX
Data Source
AI summary
A microscopy method for quantifying a fluorescence of protoporphyrin IX includes: imaging an object region onto a first detector field and a second detector field, wherein a first optical filter and a second optical filter, respectively, are arranged in the beam paths between the object region and the detector fields, the first optical filter and second optical filter respectively having a wavelength-dependent transmission characteristic; exciting at least a first fluorescence of protoporphyrin IX and a second fluorescence; recording first images and second images; and determining a spatially dependent fluorescence intensity of the first fluorescence in the object region by virtue of determining values representing a fluorescence intensity at locations in the object region, wherein the values are determined on the basis of the radiation intensities of the two detector fields detected in a spatially dependent manner and the spatially dependent wavelength-dependent detection efficiencies of the two detector fields.