Research Papers: Imaging

Direct comparison between confocal and multiphoton microscopy for rapid histopathological evaluation of unfixed human breast tissue

[+] Author Affiliations
Tadayuki Yoshitake, Michael G. Giacomelli, Lucas C. Cahill, James G. Fujimoto

Massachusetts Institute of Technology, Department of Electrical Engineering and Computer Science and Research Laboratory of Electronics, 32 Vassar Street, Cambridge, Massachusetts 02139, United States

Daniel B. Schmolze, Hilde Vardeh, Beverly E. Faulkner-Jones, James L. Connolly

Harvard Medical School, Department of Pathology, Beth Israel Deaconess Medical Center, 330 Brookline Avenue, Boston, Massachusetts 02215, United States

J. Biomed. Opt. 21(12), 126021 (Dec 29, 2016). doi:10.1117/1.JBO.21.12.126021
History: Received October 12, 2016; Accepted December 5, 2016
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Abstract.  Rapid histopathological examination of surgical specimen margins using fluorescence microscopy during breast conservation therapy has the potential to reduce the rate of positive margins on postoperative histopathology and the need for repeat surgeries. To assess the suitability of imaging modalities, we perform a direct comparison between confocal fluorescence microscopy and multiphoton microscopy for imaging unfixed tissue and compare to paraffin-embedded histology. An imaging protocol including dual channel detection of two contrast agents to implement virtual hematoxylin and eosin images is introduced that provides high quality imaging under both one and two photon excitation. Corresponding images of unfixed human breast tissue show that both confocal and multiphoton microscopy can reproduce the appearance of conventional histology without the need for physical sectioning. We further compare normal breast tissue and invasive cancer specimens imaged at multiple magnifications, and assess the effects of photobleaching for both modalities using the staining protocol. The results demonstrate that confocal fluorescence microscopy is a promising and cost-effective alternative to multiphoton microscopy for rapid histopathological evaluation of ex vivo breast tissue.

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© 2016 Society of Photo-Optical Instrumentation Engineers

Citation

Tadayuki Yoshitake ; Michael G. Giacomelli ; Lucas C. Cahill ; Daniel B. Schmolze ; Hilde Vardeh, et al.
"Direct comparison between confocal and multiphoton microscopy for rapid histopathological evaluation of unfixed human breast tissue", J. Biomed. Opt. 21(12), 126021 (Dec 29, 2016). ; http://dx.doi.org/10.1117/1.JBO.21.12.126021


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