Fresh Digital Pathology Imaging for Rapid H&E Virtual Sectioning
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Solution Overview
Problem
Existing digital pathology methods for intraoperative tumor assessment (ITA) are labor-intensive, time-consuming, prone to artifacts, and require physical sectioning, and do not achieve the necessary resolution and stitching accuracy for reliable histopathological assessment of fresh specimens using standard H&E dyes without additional training.
Innovation Solution
A rapid fresh digital-pathology (RFP) method utilizing multimodal nonlinear optical laser-raster-scanning for optical virtual sectioning, enabling high-resolution imaging of HE-stained specimens with a laser-raster-scanning field-of-view of at least 1 mm² and effective digital resolution of less than 1 micron, achieving real-time digital display and artifact-free stitching.
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 per round, but the process becomes labor intensive and limits feasible rounds of ITAs
Solution Approach 1:
The patent replaces the mechanical cryosectioning process with optical virtual sectioning using nonlinear optical laser-raster-scanning. This substitution eliminates the need for physical sectioning equipment and manual cryostat operation, thereby reducing labor intensity while maintaining rapid assessment capability
Solution Approach 2:
The patent creates optical copies of tissue structures through nonlinear optical imaging signals (two-photon excitation fluorescence and third harmonic generation) without physical sectioning. This optical copying approach preserves diagnostic information while eliminating the labor-intensive mechanical sectioning process
2Reliability
If physical sectioning is performed for FS/FFPE pathology, then histopathological assessment can be performed, but the process is time-consuming and prone to artifacts
Solution Approach 1:
The patent replaces mechanical sectioning with optical virtual sectioning using multiphoton microscopy. This eliminates physical sectioning artifacts while maintaining histopathological assessment reliability through optical sectioning of intact fresh specimens
Solution Approach 2:
The patent performs rapid tissue staining (RTS) with hematoxylin and eosin dyes before imaging, allowing fresh specimens to be prepared and stained in less than 8 minutes. This preliminary staining action enables reliable H&E-based assessment without time-consuming fixation and sectioning processes
3Productivity
If optical virtual sectioning is used for fast ITA, then assessment speed is improved, but resolution and stitching accuracy may not meet WSI standards
Solution Approach 1:
The patent uses tunable laser wavelengths (1000-1100 nm or 1200-1300 nm) and adjusts numerical aperture (NA ≥ 0.7) to optimize both imaging speed and resolution. This parameter optimization enables gigapixel-resolution imaging with effective digital lateral resolution of less than 1 micron while maintaining rapid assessment capability
Solution Approach 2:
The patent implements multi-tile combined cumulative imaging with artifact-free stitching algorithms to extend the field-of-view from single-tile 1 mm² to multi-tile 400 mm² coverage. This dimensional extension maintains gigapixel resolution across large areas while enabling comprehensive tissue assessment
4Measurement precision
If high NA objective lens is used for high resolution imaging, then digital resolution is improved, but field-of-view is limited to less than 1 mm² requiring stitching
Solution Approach 1:
The patent transitions from single-field-of-view imaging to multi-tile cumulative imaging by systematically stitching multiple 1 mm² tiles into a comprehensive 400 mm² map. This dimensional approach maintains high resolution (less than 1 micron) while dramatically expanding the observable area
Solution Approach 2:
The patent divides the large-area tissue sample into multiple manageable 1 mm² tiles that are imaged separately with high NA objectives and then computationally stitched together. This segmentation strategy enables high-resolution imaging of extensive tissue areas that would be impossible to capture in a single field-of-view
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
The RFP method provides a fast, accurate, and reliable histopathological assessment of fresh specimens in less than 8 minutes, maintaining gigapixel resolution and compatibility with standard H&E dyes, with 100% tumor identification success and FS/FFPE-comparable accuracy.
Implementation Method 1
an optical virtual sectioning via a multimodal nonlinear optical laser-raster-scanning to provide with a nonlinear multi-harmonic generation
Implementation Method 2
nonlinear multi-photon excitation fluorescence signal(s) for multichannel digitization
Data Source
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 multi-photon 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).