Real-time histological evaluation of gastrointestinal tissue using non-linear microscopy

Area of Science:

• Histopathology
• Optical Imaging
• Digital Pathology

Background:

• Optical sectioning technologies are crucial for histology, enabling real-time intraoperative assessment and efficient tissue triage.
• Digital pathology demands high-throughput optical sectioning to replace traditional slide preparation and scanning.
• Non-linear microscopy (NLM) shows promise for histology but hasn't been applied to gastrointestinal tissue.

Purpose of the Study:

• Extend NLM applications to gastrointestinal histology.
• Develop an NLM-based image atlas for automated digital pathology.
• Evaluate NLM's potential in gastrointestinal tissue assessment.

Main Methods:

• Applied NLM to gastrointestinal tissue samples.
• Developed a comprehensive NLM image atlas.
• Utilized time-lapse videos to capture 3D structures.

Main Results:

• NLM produced diagnostic-quality images comparable to H&E slides.
• NLM provided valuable 3D spatial information for assessing invasion depth, lymphovascular/perineural invasion, tumor budding, and margins.
• NLM captured 3D histological structures up to 100 µm depth.

Conclusions:

• NLM is a viable optical sectioning platform for gastrointestinal histology.
• NLM offers diagnostic-quality imaging and advanced 3D visualization.
• An NLM-based atlas can redefine anatomical pathology and advance digital pathology image analysis.