Virtual Hematoxylin and Eosin Transillumination Microscopy Using Epi-Fluorescence Imaging.

Michael G Giacomelli,Hilde Vardeh,Lennart Husvogt,Joachim Hornegger,James G Fujimoto,James L Connolly,Beverly E Faulkner-Jones,Thomas Abraham

doi:10.1371/journal.pone.0159337

Abstract

We derive a physically realistic model for the generation of virtual transillumination, white light microscopy images using epi-fluorescence measurements from thick, unsectioned tissue. We demonstrate this technique by generating virtual transillumination H&E images of unsectioned human breast tissue from epi-fluorescence multiphoton microscopy data. The virtual transillumination algorithm is shown to enable improved contrast and color accuracy compared with previous color mapping methods. Finally, we present an open source implementation of the algorithm in OpenGL, enabling real-time GPU-based generation of virtual transillumination microscopy images using conventional fluorescence microscopy systems.

Highlights

Classical histopathology is the gold standard for clinical evaluation of many suspected neoplasms, including cancer of the breast
The vertex shader defines a surface on which a virtual transillumination microscopy image is rendered and provides features such as scaling of image data to the monitor resolution and rotation or mirroring of the image data to match the physical geometry of the microscope
To demonstrate the virtual transillumination algorithm, we present images from discarded surgical breast mastectomy specimens from a patient undergoing mastectomy for invasive ductal carcinoma

Summary

Introduction

Classical histopathology is the gold standard for clinical evaluation of many suspected neoplasms, including cancer of the breast. The tissue specimen is fixed, dehydrated, paraffin embedded, sectioned into micron thin slices, and visualized using molecularly-specific absorptive dyes in a transillumination light microscope. The complex specimen preparation requires processing times on the order of one day, which precludes using conventional histopathology for realtime applications such as surgical or biopsy guidance [1]. Repeat procedures pose additional risk to patients, may delay adjuvant therapy, reduce cosmetic outcomes, and impose an additional financial burden on the healthcare system [5,6]. To address this problem, various groups have investigated fluorescence microscopy techniques such as confocal microscopy [7,8], multiphoton microscopy

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Results

Conclusion