Fluorescence-Based Tissue Localization in FFPE Blocks
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Solution Overview
Problem
Existing methods for analyzing formalin-fixed paraffin-embedded (FFPE) tissue blocks are inaccurate and inconsistent in distinguishing tissue from paraffin, due to sensitivity to optical and surface characteristics, making it difficult to determine the location of tissue in the embedding medium.
Innovation Solution
The method involves irradiating the embedded sample with light of specific wavelengths (200 nm to 600 nm) to detect fluorescence emission from endogenous fluorophores, allowing for the accurate determination of tissue location within the FFPE block using fluorescence emission patterns.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If automated analysis using camera and light source is used to image tissue block surface, then productivity is improved, but measurement precision deteriorates due to sensitivity to optical and surface characteristics variability
Solution Approach 1:
The patent changes the detection parameter from reflected light intensity to fluorescence emission wavelength. By using fluorescence excitation (200-600 nm) and detecting emitted fluorescence, the system achieves wavelength-specific detection that is insensitive to surface texture and optical property variations, thereby maintaining high measurement precision while preserving automated analysis efficiency
Solution Approach 2:
The patent replaces the optical reflection-based detection system with a fluorescence-based detection system. This substitution eliminates dependence on surface characteristics and optical properties by utilizing the intrinsic fluorescence properties of tissue components, achieving both automated operation and high precision tissue location determination
2Measurement precision
If technician manually examines FFPE block to observe diffuse tissue image, then measurement precision is improved, but productivity deteriorates due to time-consuming manual analysis
Solution Approach 1:
The patent replaces manual visual examination with automated fluorescence detection. The system uses light sources to excite fluorescence in tissue components and detectors to capture emission patterns, automatically determining tissue boundaries with precision comparable to manual examination while dramatically increasing processing speed
Solution Approach 2:
The patent introduces fluorescence emission as an intermediary signal between the tissue sample and the detection system. This fluorescence signal provides contrast based on molecular composition rather than surface appearance, enabling automated systems to achieve the same measurement precision as manual examination by detecting tissue-specific fluorescence patterns
3Ease of operation
If existing optical methods are used to distinguish tissue from paraffin, then ease of operation is maintained, but measurement precision deteriorates when tissue is optically indistinguishable from embedding medium
Solution Approach 1:
The patent changes the detection approach from reflected light intensity (which depends on surface properties) to fluorescence emission wavelength (which depends on molecular composition). This parameter change enables differentiation of tissue from paraffin based on their distinct fluorescence spectra, maintaining operational simplicity while achieving high differentiation accuracy even when surface properties are similar
Solution Approach 2:
The patent utilizes fluorescence emission wavelengths (color) as the distinguishing feature between tissue and paraffin. By detecting the specific wavelengths of fluorescence emitted by tissue components versus paraffin, the system achieves accurate differentiation based on molecular composition rather than surface appearance, preserving ease of operation while improving measurement precision
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 provides a consistent and accurate method for differentiating tissue from paraffin, enabling precise localization of tissue in FFPE samples, even when they are optically indistinguishable from the embedding medium, and facilitates accurate sectioning for further analysis.
Implementation Method 1
irradiating an embedded sample with light having a wavelength of from about 200 nm to about 600 nm, wherein the embedded sample comprises a tissue and an embedding medium; detecting fluorescence emission of the embedded sample
Data Source
Figure 1A~1B
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Figure 4A~4B
AI summary
The present disclosure is directed to an improved method and device for distinguishing tissue from an embedding medium, such as paraffin in a formalin-fixed paraffin-embedded sample. The method involves the use of fluorescence of naturally-occurring species in tissue to determine the location of the tissue in the embedded sample. For example, an embedded sample (703) is generally excited by light of a selected wavelength from an LED light source (707), and the fluorescence emission at an emitted wavelength is recorded by a camera (717) and is used to locate the boundary or location of the tissue in the embedded sample. The device may a include microtome blade (705), e.g. for slicing a section from the embedded sample after determination of the location of the tissue.