Nonlinear Optical Microscopy for Label-Free Immune Cell Imaging in Human Skin
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current imaging techniques are limited in their ability to non-invasively image immune cells beneath the skin surface, distinguish immune cells based on morphology, metabolic state, and behavior, and cover large areas rapidly, which is crucial for understanding autoimmune diseases and evaluating immunotherapy responses.
Innovation Solution
A nonlinear optical microscopy system, such as a fast, large area multiphoton exoscope (FLAME), is used to generate depth-resolved images with sub-micron resolution, analyzing fluorescence lifetime signals of endogenous biomolecules like NADH to distinguish immune cell populations based on morphological, metabolic, and behavioral signatures.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If current imaging techniques are used to image immune cells beneath skin surface, then imaging can be performed, but the ability to distinguish immune cells based on morphology, metabolic state, and behavior is limited
Solution Approach 1:
The patent segments the imaging process into multiple specialized modules: multiphoton excitation for deep tissue penetration, fluorescence lifetime imaging for metabolic state detection, and computational analysis for cell classification. This segmentation allows each module to optimize for its specific function while achieving comprehensive immune cell characterization
Solution Approach 2:
The patent utilizes fluorescence lifetime as an additional parameter beyond intensity to distinguish immune cell populations. By measuring the temporal decay characteristics of fluorescent signals from endogenous biomolecules like NADH and FAD, the system can differentiate metabolic states without requiring exogenous labels, thereby improving measurement precision
2Productivity
If imaging techniques attempt to cover large areas rapidly, then imaging speed improves, but the ability to retrieve detailed metabolic information is compromised
Solution Approach 1:
The patent employs periodic scanning patterns that systematically cover large skin areas while allocating sufficient dwell time at each pixel to capture complete fluorescence decay curves. The structured illumination and scanning approach ensures that metabolic information is not lost despite the need for rapid coverage
Solution Approach 2:
The system maintains continuous fluorescence signal acquisition during scanning, using real-time temporal binning to accumulate photons across multiple scan passes. This continuous action ensures that even rapidly acquired images retain sufficient signal-to-noise ratio for metabolic signature extraction
3Ease of operation
If non-invasive imaging of immune cells is performed, then patient comfort is maintained, but the resolution and depth capability are limited
Solution Approach 1:
The patent replaces mechanical biopsy or surgical intervention with optical field-based multiphoton microscopy. By using nonlinear optical interactions that are inherently confined to the focal volume, the system achieves high resolution imaging at depth without physical contact or tissue disruption, maintaining non-invasive operation while preserving image quality
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 rapid, non-invasive identification and quantification of immune cell populations in human skin, facilitating clinical applications like diagnosing skin disorders, understanding autoimmunity, and evaluating immunotherapy responses.
Implementation Method 1
a nonlinear optical imaging system that generates depth-resolved images over large areas with sub-micron resolution based on fluorescence signals generated from one or more endogenous biomolecules found in human skin
Implementation Method 2
fluorescence signals generated from one or more endogenous biomolecules found in human skin, wherein one of the endogenous biomolecules is the reduced form of nicotinamide adenine dinucleotide (NADH)
Implementation Method 3
analyzing the image(s) generated from the nonlinear optical imaging system to retrieve information about the metabolic signatures of the detected immune cells by temporal binning the decays of fluorescence lifetime signals
Data Source
AI summary
The disclosure provides noninvasive methods for the in vivo identification and characterization of various immune cell populations in human skin. The methods of the disclosure employ advanced imaging systems based on nonlinear optical microscopy to generate images that are analyzed to detect, identify, differentiate, and quantify immune cell populations in human skin by their morphological, metabolic and behavioral signatures.