Microscopy with ultraviolet surface excitation for wide-area pathology of breast surgical margins

Abstract. Intraoperative assessment of breast surgical margins will be of value for reducing the rate of re-excision surgeries for lumpectomy patients. While frozen-section histology is used for intraoperative guidance of certain cancers, it provides limited sampling of the margin surface (typically <1  %   of the margin) and is inferior to gold-standard histology, especially for fatty tissues that do not freeze well, such as breast specimens. Microscopy with ultraviolet surface excitation (MUSE) is a nondestructive superficial optical-sectioning technique that has the potential to enable rapid, high-resolution examination of excised margin surfaces. Here, a MUSE system is developed with fully automated sample translation to image fresh tissue surfaces over large areas and at multiple levels of defocus, at a rate of ∼5  min  /  cm2. Surface extraction is used to improve the comprehensiveness of surface imaging, and 3-D deconvolution is used to improve resolution and contrast. In addition, an improved fluorescent analog of conventional H&E staining is developed to label fresh tissues within ∼5  min for MUSE imaging. We compare the image quality of our MUSE system with both frozen-section and conventional H&E histology, demonstrating the feasibility to provide microscopic visualization of breast margin surfaces at speeds that are relevant for intraoperative use.

[1]  Andrew B. Sholl,et al.  Gigapixel surface imaging of radical prostatectomy specimens for comprehensive detection of cancer-positive surgical margins using structured illumination microscopy , 2016, Scientific Reports.

[2]  S. Griffey,et al.  The accuracy of intraoperative diagnoses based on examination of frozen sections. A prospective comparison with paraffin-embedded sections. , 1993, Veterinary surgery : VS.

[3]  J. Fujimoto,et al.  Rapid imaging of surgical breast excisions using direct temporal sampling two photon fluorescent lifetime imaging. , 2015, Biomedical optics express.

[4]  R. Pleijhuis,et al.  Obtaining Adequate Surgical Margins in Breast-Conserving Therapy for Patients with Early-Stage Breast Cancer: Current Modalities and Future Directions , 2009, Annals of Surgical Oncology.

[5]  Monica Morrow,et al.  Should intraoperative frozen section evaluation of breast lumpectomy margins become routine practice? , 2012, American journal of clinical pathology.

[6]  M. Osborne,et al.  The influence of additional surgical margins on the total specimen volume excised and the reoperative rate after breast-conserving surgery. , 2006, American journal of surgery.

[7]  Katherine N. Elfer,et al.  DRAQ5 and Eosin (‘D&E’) as an Analog to Hematoxylin and Eosin for Rapid Fluorescence Histology of Fresh Tissues , 2016, PloS one.

[8]  Kelly K. Hunt,et al.  Role for Intraoperative Margin Assessment in Patients Undergoing Breast-Conserving Surgery , 2007, Annals of Surgical Oncology.

[9]  James G. Fujimoto,et al.  Assessment of breast pathologies using nonlinear microscopy , 2014, Proceedings of the National Academy of Sciences.

[10]  Alan I Glaser,et al.  Survival after lumpectomy and mastectomy for early stage invasive breast cancer: The effect of age and hormone receptor status , 2013, Cancer.

[11]  A. Degnim,et al.  Reasons for Re-Excision After Lumpectomy for Breast Cancer: Insight from the American Society of Breast Surgeons MasterySM Database , 2014, Annals of Surgical Oncology.

[12]  Stephan Saalfeld,et al.  Globally optimal stitching of tiled 3D microscopic image acquisitions , 2009, Bioinform..

[13]  Peter A Ubel,et al.  Effect of esthetic outcome after breast-conserving surgery on psychosocial functioning and quality of life. , 2008, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[14]  O. Carrasco-Zevallos,et al.  Review of intraoperative optical coherence tomography: technology and applications [Invited]. , 2017, Biomedical optics express.

[15]  Gerrit-Jan Liefers,et al.  Survival after lumpectomy and mastectomy for early stage invasive breast cancer: The effect of age and hormone receptor status , 2013, Cancer.

[16]  J. Lewis,et al.  Frozen section analysis of margins for head and neck tumor resections: reduction of sampling errors with a third histologic level , 2011, Modern Pathology.

[17]  L. Jones,et al.  Characterization of SYBR Gold nucleic acid gel stain: a dye optimized for use with 300-nm ultraviolet transilluminators. , 1999, Analytical biochemistry.

[18]  Stephen A. Boppart,et al.  Stain-free histopathology by programmable supercontinuum pulses , 2016, Nature Photonics.

[19]  W C Maccarty.  The Diagnostic Reliability of Frozen Sections. , 1929, The American journal of pathology.

[20]  Tadayuki Yoshitake,et al.  Multiscale nonlinear microscopy and widefield white light imaging enables rapid histological imaging of surgical specimen margins. , 2018, Biomedical optics express.

[21]  B. E. F. Isher,et al.  Twenty-year follow-up of a randomized trial comparing total mastectomy, lumpectomy, and lumpectomy plus irradiation for the treatment of invasive breast cancer. , 2002 .

[22]  Lawrence D. True,et al.  Light-sheet microscopy for slide-free non-destructive pathology of large clinical specimens , 2017, Nature Biomedical Engineering.

[23]  Tadayuki Yoshitake,et al.  Rapid histopathological imaging of skin and breast cancer surgical specimens using immersion microscopy with ultraviolet surface excitation , 2018, Scientific Reports.

[24]  M. Trivella,et al.  Reoperation rates after breast conserving surgery for breast cancer among women in England: retrospective study of hospital episode statistics , 2012, BMJ : British Medical Journal.

[25]  Tyler C. Schlichenmeyer,et al.  High-Resolution Rapid Diagnostic Imaging of Whole Prostate Biopsies Using Video-Rate Fluorescence Structured Illumination Microscopy. , 2015, Cancer research.

[26]  Chi Zhang,et al.  Fast label-free multilayered histology-like imaging of human breast cancer by photoacoustic microscopy , 2017, Science Advances.

[27]  Farzad Fereidouni,et al.  Microscopy with ultraviolet surface excitation (MUSE): A novel approach to real‐time inexpensive slide‐free dermatopathology , 2018, Journal of cutaneous pathology.

[28]  Zachary T. Harmany,et al.  Microscopy with ultraviolet surface excitation for rapid slide-free histology , 2017, Nature Biomedical Engineering.

[29]  Nathan D. Shemonski,et al.  Real-time Imaging of the Resection Bed Using a Handheld Probe to Reduce Incidence of Microscopic Positive Margins in Cancer Surgery. , 2015, Cancer research.

[30]  G. Zanghirati,et al.  Towards real-time image deconvolution: application to confocal and STED microscopy , 2013, Scientific Reports.

[31]  Guillermo J Tearney,et al.  Spectrally encoded confocal microscopy for diagnosing breast cancer in excision and margin specimens , 2016, Laboratory Investigation.

[32]  Joachim Hornegger,et al.  Virtual Hematoxylin and Eosin Transillumination Microscopy Using Epi-Fluorescence Imaging , 2016, PloS one.

[33]  Marc A Bruce,et al.  Real-time GPU-based 3D Deconvolution. , 2013, Optics express.

[34]  Miguel Cordova,et al.  Intraoperative imaging during Mohs surgery with reflectance confocal microscopy: initial clinical experience , 2015, Journal of biomedical optics.

[35]  Saurabh Sinha,et al.  Intravital imaging by simultaneous label-free autofluorescence-multiharmonic microscopy , 2018, Nature Communications.

[36]  Katherine N. Elfer,et al.  Video-rate structured illumination microscopy for high-throughput imaging of large tissue areas. , 2014, Biomedical optics express.

[37]  Michael Unser,et al.  Complex wavelets for extended depth‐of‐field: A new method for the fusion of multichannel microscopy images , 2004, Microscopy research and technique.

[38]  Milind Rajadhyaksha,et al.  Confocal microscopy with strip mosaicing for rapid imaging over large areas of excised tissue , 2013, Journal of biomedical optics.