Rapid Fresh Digital Pathology via Nonlinear Optical Laser Scanning
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Solution Overview
Problem
Current digital pathology methods for intraoperative tumor assessment are labor-intensive, artifact-prone, and lack the speed and resolution required for reliable, real-time evaluation of fresh specimens stained with hematoxylin and eosin dyes, particularly for large-field imaging and gigapixel sampling.
Innovation Solution
A rapid fresh digital-pathology method using multimodal nonlinear optical laser-raster-scanning for optical virtual sectioning, enabling high-resolution imaging with a large field-of-view and real-time digital display of histopathological features without physical sectioning or fixation, utilizing a fiber-based or chromium-forsterite pulsed laser source and achieving gigapixel sampling in under 8 minutes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Loss of time
If frozen section pathology is used for intraoperative tumor assessment, then assessment time is reduced to 30 minutes, but the process becomes labor intensive and artifact-prone
Solution Approach 1:
The patent replaces mechanical cryosectioning with optical virtual sectioning using nonlinear optical laser-raster-scanning. The laser system optically sections the fresh specimen without physical cutting, eliminating the need for cryostats, microtomes, and manual sectioning operations while maintaining rapid assessment capability
Solution Approach 2:
The patent changes the physical state parameter of the specimen from frozen to fresh, and the imaging modality from transmitted light microscopy to nonlinear optical laser scanning. This allows optical sectioning of fresh, unfixed, unstained tissue with preserved histological architecture, eliminating artifacts associated with freezing and sectioning
2Measurement precision
If optical virtual sectioning with hematoxylin staining is used, then high-contrast cell nuclei visualization is achieved, but nonlinear multi-harmonic generation is required which complicates the system
Solution Approach 1:
The patent employs a unified nonlinear optical laser-raster-scanning system that simultaneously performs optical sectioning, hematoxylin-specific imaging via third harmonic generation, and eosin-specific imaging via two-photon excited fluorescence. This multi-functional approach achieves high-contrast nuclei visualization without requiring separate specialized systems for each imaging mode
Solution Approach 2:
The patent utilizes the composite staining properties of H&E dyes, where hematoxylin provides nuclear contrast through third harmonic generation and eosin provides cytoplasmic contrast through two-photon fluorescence. The nonlinear optical system detects both signals simultaneously, achieving comprehensive tissue visualization with enhanced nuclei contrast
3Measurement precision
If gigapixel sampling with Nyquist-satisfied resolution is achieved, then diagnostic reliability is maintained, but imaging speed and real-time display capability are compromised
Solution Approach 1:
The patent uses resonant galvanometric scanners that operate at resonant frequencies (e.g., 8 kHz line rate) to perform rapid periodic raster scanning across the specimen. This periodic high-speed scanning, combined with persistent buffer accumulation and real-time digital display, achieves gigapixel sampling at Nyquist resolution while maintaining imaging speed under 8 minutes for 1 cm² areas
Solution Approach 2:
The patent implements dynamic scanning with variable line rates and real-time image reconstruction. The resonant scanner dynamically adjusts scanning parameters while the system accumulates data in persistent buffers and performs real-time digital display, enabling adaptive optimization between resolution and speed based on diagnostic requirements
4Loss of information
If traditional WSI methods are used, then digital archiving is achieved, but physical sectioning and fixation are required which take 1-2 days
Solution Approach 1:
The patent replaces mechanical sectioning and chemical fixation processes with optical virtual sectioning of fresh specimens. The nonlinear optical laser system creates digital images of intact, unfixed tissue, eliminating the 1-2 day processing workflow while maintaining digital archiving capability through gigapixel sampling at Nyquist-satisfied resolution
Solution Approach 2:
The patent performs rapid hematoxylin and eosin staining (3-5 minutes) on fresh specimens before imaging, which preserves tissue architecture and provides diagnostic contrast without requiring subsequent fixation and sectioning. This preliminary staining action enables direct optical sectioning and digital archiving, dramatically reducing total processing time
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 method provides a 4 times faster assessment with high-resolution imaging and 100% tumor identification accuracy, comparable to formalin-fixed paraffin-embedded biopsies, while maintaining diagnostic reliability and eliminating the need for deep learning or additional interpretation training.
Implementation Method 1
nonlinear multi-harmonic generation approach to visualize the high-contrast cell nuclei
Implementation Method 2
two-photon excited fluorescence signal for multichannel digitization and real-time digital display
Data Source
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AI summary
A rapid fresh digital-pathology (RFP) method for assessing an excised unfixed biological specimen stained with hematoxylin (H) or eosin (E) or both hematoxylin and eosin (HE) staining dyes. The RFP method is assisted by a rapid tissue staining (RTS) procedure which is performed on the excised unfixed biological specimen, involving a short fixation; an H-staining; a rinsing; a bluing; an E-staining; a rinsing; and finally, a covering of a stained specimen with a coverslip. The RFP method is further assisted by a multimodal nonlinear optical laser-raster-scanning approach to provide with a nonlinear multi-harmonic generation and/or a nonlinear multiphoton excitation fluorescence signal(s) for multichannel digitization and real-time digital display of H- or E- or HE-specific histopathological features while providing a centimeter-scale imaging area, a submicron digital resolution, and a sustained effective data throughput of at least 500 Megabits per second (Mbps).