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An end-to-end workflow for nondestructive 3D pathology.

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This study presents a detailed 3D pathology workflow for high-quality tissue imaging. The protocol, using fluorescent H&E staining and light-sheet microscopy, enables comprehensive volumetric analysis for research.

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Area of Science:

  • Pathology and Laboratory Medicine
  • Biomedical Imaging
  • Histology

Background:

  • Conventional 2D pathology methods have limitations in tissue imaging volume and contextual information.
  • Three-dimensional (3D) pathology offers enhanced tissue visualization without destructive sectioning.
  • Generating consistent, high-quality 3D pathology datasets requires meticulous optimization of the entire workflow.

Purpose of the Study:

  • To provide a detailed, end-to-end protocol for a 3D pathology workflow.
  • To enable consistent generation of high-quality 3D pathology datasets for preclinical and clinical studies.
  • To offer guidelines for diverse end-users, including biologists, clinical researchers, and engineers.

Main Methods:

  • Development and description of a comprehensive 3D pathology workflow.
  • Utilized light-sheet microscopy as an illustrative imaging platform.
  • Focused on a fluorescent analog of hematoxylin and eosin (H&E) staining for compatibility with gold-standard pathological reports.

Main Results:

  • An end-to-end 3D pathology procedure is presented, covering tissue preparation, imaging, and initial data processing.
  • The protocol is detailed sufficiently for well-controlled preclinical and clinical studies.
  • The workflow requires 3-6 days for completion, excluding data analysis time.

Conclusions:

  • The provided protocol facilitates the generation of high-quality 3D pathology datasets.
  • This workflow enhances volumetric tissue analysis capabilities beyond conventional 2D methods.
  • The guidelines support broader adoption of 3D pathology in research and clinical settings.