PRM-SRS Imaging for Lipid Subtype Identification
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Solution Overview
Problem
Current lipidomic technologies, such as shotgun lipidomics and MALDI-MS imaging, are destructive and lack the ability to distinguish multiple lipid subtypes in cells and tissues with high chemical specificity, while optical imaging techniques struggle to differentiate diverse molecular species due to limited spectral resolution and reliance on markers or labels.
Innovation Solution
The development of a multi-molecular hyperspectral Penalized Reference Matching algorithm with Stimulated Raman Scattering (PRM-SRS) imaging, which enables faster data processing and direct user-defined visualization by quantifying spectral similarity between image pixel spectra and known reference spectra, reducing false positives through positional discrepancy penalties.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If mass spectrometry or nuclear magnetic resonance techniques are used for lipid detection, then sensitivity and detection speed are improved, but the sample is destroyed during the imaging process
Solution Approach 1:
The patent replaces mass spectrometry and nuclear magnetic resonance techniques with stimulated Raman scattering (SRS) microscopy, which uses optical fields instead of mechanical or electromagnetic fields that cause sample destruction. This substitution enables non-destructive imaging while maintaining high detection speed and sensitivity through the use of laser-induced Raman scattering signals.
Solution Approach 2:
The patent utilizes the phase transition properties of lipids by detecting Raman scattering signals that are sensitive to molecular vibrations and phase states. This allows differentiation of lipid subtypes in various phases (liquid crystalline, gel, solid) without destroying the sample, achieving both non-destructive imaging and molecular specificity.
2Manufacturing precision
If conventional MALDI-MS imaging is used for lipid visualization, then label-free imaging is achieved, but the lateral resolution is limited to cell diameter scale and sample is destroyed
Solution Approach 1:
The patent replaces MALDI-MS imaging with SRS microscopy, substituting ionization-based detection with Raman scattering-based optical detection. This enables subcellular resolution imaging at the scale of 200-500 nm laterally and 1-5 µm axially, while maintaining non-destructive sample imaging through the use of coherent Raman scattering signals that do not require sample destruction or ionization.
3Object-affected harmful factors
If labeled optical imaging techniques are used for lipid visualization, then non-destructive imaging is achieved, but the ability to differentiate diverse molecular species is limited due to marker availability and native distribution alteration
Solution Approach 1:
The patent replaces fluorescence labeling with Raman scattering-based detection, which inherently provides molecular specificity through vibrationally resonant signals that are unique to each molecular species. This eliminates the need for external markers or labels, avoiding alterations to native lipid distribution while maintaining non-destructive imaging and achieving high molecular specificity through the fingerprint region of Raman spectra.
Solution Approach 2:
The patent utilizes the intrinsic Raman scattering properties of lipid molecules themselves for identification and differentiation, without requiring external labels or markers. Each lipid subtype serves as its own identifier through its unique Raman spectral signature, enabling label-free, non-destructive, and highly specific molecular differentiation.
4Measurement precision
If hyperspectral imaging with high spectral resolution is used for lipid species differentiation, then chemical specificity is improved, but imaging time becomes prohibitively long
Solution Approach 1:
The patent employs periodic modulation of the pump and Stokes laser beams at different frequencies, using lock-in detection to selectively detect specific Raman shifts. This frequency-modulated approach enables rapid acquisition of hyperspectral data by multiplexing multiple spectral measurements within a single imaging frame, significantly reducing total imaging time while maintaining high spectral resolution and chemical specificity.
Solution Approach 2:
The patent uses dynamic frequency tuning of the laser beams to rapidly scan through different Raman shift frequencies, enabling real-time hyperspectral imaging. The system dynamically adjusts laser frequencies and detection parameters to capture the full spectral range needed for lipid differentiation, achieving both high spectral resolution and fast imaging speeds through adaptive, time-efficient measurement sequences.
5Measurement precision
If unsupervised unmixing methods like MCR-ALS are used for lipid species identification, then spectral decomposition is achieved, but computational cost is high and direct molecular identification is not possible
Solution Approach 1:
The patent replaces complex computational unmixing algorithms with direct spectral matching against a database of reference Raman spectra. By comparing measured hyperspectral pixels with pre-acquired reference spectra of known lipid species, the system directly identifies molecular species without requiring computationally intensive iterative decomposition, significantly reducing computational complexity while maintaining high spectral resolution and accurate molecular identification.
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 allows for the efficient identification and visualization of multiple lipid subtypes in cells and tissues with high chemical specificity and subcellular resolution, enhancing the ability to understand lipid metabolism and its role in diseases, and can be applied to various biological samples with improved accuracy and speed compared to existing methods.
Implementation Method 1
Stimulated Raman scattering (SRS) microscopy has demonstrated advantages of non-destructive 3D imaging with subcellular resolution in a label-free manner
Implementation Method 2
PRM-SRS can advantageously provide faster data processing and direct user-defined visualization with enhanced chemical specificity for distinguishing clinically relevant lipid subtypes in different organs and species
Data Source
AI summary
Multi-molecular hyperspectral PRM-SRS imaging technologies advantageously provide faster data processing and direct user-defined visualization with enhanced chemical specificity for distinguishing clinically relevant lipid subtypes in different organs and species. PRM-SMS may include quantifying the spectral similarity between an image pixel spectrum and a known reference spectrum.