title: Direct comparison between confocal and multiphoton microscopy for rapid histopathological evaluation of unfixed human breast tissue.
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.
authors.affiliation: Massachusetts Institute of Technology, Department of Electrical Engineering and Computer Science and Research Laboratory of Electronics, 32 Vassar Street, Cambridge, Massachusetts 02139, United States.
authors.forename: Tadayuki
authors.initials: T
authors.lastname: Yoshitake
authors.affiliation: Massachusetts Institute of Technology, Department of Electrical Engineering and Computer Science and Research Laboratory of Electronics, 32 Vassar Street, Cambridge, Massachusetts 02139, United States.
authors.forename: Michael G
authors.initials: MG
authors.lastname: Giacomelli
authors.affiliation: Massachusetts Institute of Technology, Department of Electrical Engineering and Computer Science and Research Laboratory of Electronics, 32 Vassar Street, Cambridge, Massachusetts 02139, United States.
authors.forename: Lucas C
authors.initials: LC
authors.lastname: Cahill
authors.affiliation: Harvard Medical School, Department of Pathology, Beth Israel Deaconess Medical Center, 330 Brookline Avenue, Boston, Massachusetts 02215, United States.
authors.forename: Daniel B
authors.initials: DB
authors.lastname: Schmolze
authors.affiliation: Harvard Medical School, Department of Pathology, Beth Israel Deaconess Medical Center, 330 Brookline Avenue, Boston, Massachusetts 02215, United States.
authors.forename: Hilde
authors.initials: H
authors.lastname: Vardeh
authors.affiliation: Harvard Medical School, Department of Pathology, Beth Israel Deaconess Medical Center, 330 Brookline Avenue, Boston, Massachusetts 02215, United States.
authors.forename: Beverly E
authors.initials: BE
authors.lastname: Faulkner-Jones
authors.affiliation: Harvard Medical School, Department of Pathology, Beth Israel Deaconess Medical Center, 330 Brookline Avenue, Boston, Massachusetts 02215, United States.
authors.forename: James L
authors.initials: JL
authors.lastname: Connolly
authors.affiliation: Massachusetts Institute of Technology, Department of Electrical Engineering and Computer Science and Research Laboratory of Electronics, 32 Vassar Street, Cambridge, Massachusetts 02139, United States.
authors.forename: James G
authors.initials: JG
authors.lastname: Fujimoto
mesh_terms: D001940:Breast / Q000000981:diagnostic imaging; D001943:Breast Neoplasms / Q000000981:diagnostic imaging*; D004867:Equipment Design; D005260:Female; D006801:Humans; D007090:Image Interpretation, Computer-Assisted / Q000379:methods*; D018613:Microscopy, Confocal / Q000295:instrumentation / Q000379:methods*; D036641:Microscopy, Fluorescence, Multiphoton / Q000295:instrumentation / Q000379:methods*