PMID- 29131161 OWN - NLM STAT- MEDLINE DCOM- 20181217 LR - 20240315 IS - 1530-0307 (Electronic) IS - 0023-6837 (Print) IS - 0023-6837 (Linking) VI - 98 IP - 1 DP - 2018 Jan TI - Rapid virtual hematoxylin and eosin histology of breast tissue specimens using a compact fluorescence nonlinear microscope. PG - 150-160 LID - 10.1038/labinvest.2017.116 [doi] AB - Up to 40% of patients undergoing breast conserving surgery for breast cancer require repeat surgeries due to close to or positive margins. The lengthy processing required for evaluating surgical margins by standard paraffin-embedded histology precludes its use during surgery and therefore, technologies for rapid evaluation of surgical pathology could improve the treatment of breast cancer by reducing the number of surgeries required. We demonstrate real-time histological evaluation of breast cancer surgical specimens by staining specimens with acridine orange (AO) and sulforhodamine 101 (SR101) analogously to hematoxylin and eosin (H&E) and then imaging the specimens with fluorescence nonlinear microscopy (NLM) using a compact femtosecond fiber laser. A video-rate computational light absorption model was used to produce realistic virtual H&E images of tissue in real time and in three dimensions. NLM imaging could be performed to depths of 100 mum below the tissue surface, which is important since many surgical specimens require subsurface evaluation due to contamination artifacts on the tissue surface from electrocautery, surgical ink, or debris from specimen handling. We validate this method by expert review of NLM images compared to formalin-fixed, paraffin-embedded (FFPE) H&E histology. Diagnostically important features such as normal terminal ductal lobular units, fibrous and adipose stromal parenchyma, inflammation, invasive carcinoma, and in situ lobular and ductal carcinoma were present in NLM images associated with pathologies identified on standard FFPE H&E histology. We demonstrate that AO and SR101 were extracted to undetectable levels after FFPE processing and fluorescence in situ hybridization (FISH) HER2 amplification status was unaffected by the NLM imaging protocol. This method potentially enables cost-effective, real-time histological guidance of surgical resections. FAU - Cahill, Lucas C AU - Cahill LC AD - Department of Electrical Engineering and Computer Science and Research Laboratory of Electronics, Massachusetts Institute of Technology, Cambridge, MA, USA. AD - Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA, USA. FAU - Giacomelli, Michael G AU - Giacomelli MG AD - Department of Electrical Engineering and Computer Science and Research Laboratory of Electronics, Massachusetts Institute of Technology, Cambridge, MA, USA. FAU - Yoshitake, Tadayuki AU - Yoshitake T AD - Department of Electrical Engineering and Computer Science and Research Laboratory of Electronics, Massachusetts Institute of Technology, Cambridge, MA, USA. FAU - Vardeh, Hilde AU - Vardeh H AD - Department of Pathology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA. FAU - Faulkner-Jones, Beverly E AU - Faulkner-Jones BE AD - Department of Pathology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA. FAU - Connolly, James L AU - Connolly JL AD - Department of Pathology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA. FAU - Sun, Chi-Kuang AU - Sun CK AD - Department of Electrical Engineering and Graduate Institute of Photonics and Optoelectronics, National Taiwan University, Taipei, Taiwan. FAU - Fujimoto, James G AU - Fujimoto JG AD - Department of Electrical Engineering and Computer Science and Research Laboratory of Electronics, Massachusetts Institute of Technology, Cambridge, MA, USA. LA - eng GR - F32 CA183400/CA/NCI NIH HHS/United States GR - R01 CA075289/CA/NCI NIH HHS/United States GR - R01 CA178636/CA/NCI NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, U.S. Gov't, Non-P.H.S. PT - Validation Study DEP - 20171113 PL - United States TA - Lab Invest JT - Laboratory investigation; a journal of technical methods and pathology JID - 0376617 RN - 0 (Coloring Agents) RN - 0 (Fluorescent Dyes) RN - 0 (Rhodamines) RN - F30N4O6XVV (Acridine Orange) RN - FX0ES3271V (sulforhodamine 101) SB - IM MH - Acridine Orange/chemistry MH - Breast/cytology/immunology/*pathology/surgery MH - Breast Carcinoma In Situ/diagnosis/immunology/*pathology/surgery MH - Breast Neoplasms/diagnosis/immunology/*pathology/surgery MH - Carcinoma, Ductal, Breast/diagnosis/immunology/*pathology/surgery MH - Carcinoma, Lobular/diagnosis/immunology/*pathology/surgery MH - Coloring Agents/chemistry MH - Female MH - Fluorescent Dyes/chemistry MH - Humans MH - Imaging, Three-Dimensional MH - Intraoperative Period MH - *Margins of Excision MH - Mastectomy MH - Mastectomy, Segmental MH - Microscopy, Fluorescence MH - Neoplasm Invasiveness MH - Nonlinear Optical Microscopy MH - Organ Sparing Treatments MH - Rhodamines/chemistry PMC - PMC5752596 MID - NIHMS905378 COIS- DISCLOSURE/CONFLICT OF INTEREST None declared. EDAT- 2017/11/14 06:00 MHDA- 2018/12/18 06:00 PMCR- 2018/05/13 CRDT- 2017/11/14 06:00 PHST- 2017/05/04 00:00 [received] PHST- 2017/09/07 00:00 [revised] PHST- 2017/09/08 00:00 [accepted] PHST- 2017/11/14 06:00 [pubmed] PHST- 2018/12/18 06:00 [medline] PHST- 2017/11/14 06:00 [entrez] PHST- 2018/05/13 00:00 [pmc-release] AID - S0023-6837(22)01160-6 [pii] AID - 10.1038/labinvest.2017.116 [doi] PST - ppublish SO - Lab Invest. 2018 Jan;98(1):150-160. doi: 10.1038/labinvest.2017.116. Epub 2017 Nov 13.