Magnetically Promoted Rapid Immunofluorescence Staining for Frozen Tissue Sections

  • J Histochem Cytochem. 2019 Aug;67(8):575-587. doi: 10.1369/0022155419841023.
Tatsuya Onishi  1  2 ,  Sachiko Matsuda  1 ,  Yuki Nakamura  1 ,  Junko Kuramoto  3 ,  Akinori Tsuruma  4 ,  Satoshi Sakamoto  4 ,  Shunichi Suzuki  5 ,  Daiichiro Fuchimoto  5 ,  Akira Onishi  6 ,  Shinichi Chikaki  7 ,  Miki Kaneko  7 ,  Akihiro Kuwahata  7 ,  Masaki Sekino  7 ,  Hiroshi Yasuno  8 ,  Naohiro Hanyu  8 ,  Tomoko Kurita  9 ,  Hiroyuki Takei  9 ,  Takashi Sakatani  10 ,  Kanae Taruno  11 ,  Seigo Nakamura  11 ,  Tetsu Hayashida  1 ,  Hiromitsu Jinno  12 ,  Moriaki Kusakabe  13  14 ,  Hiroshi Handa  15 ,  Kaori Kameyama  16 ,  Yuko Kitagawa  1
Affiliations
  • 1. Department of Surgery, Keio University School of Medicine, Tokyo, Japan.
  • 2. Department of Breast Surgery, National Cancer Center Hospital East, Kashiwa, Japan.
  • 3. Department of Pathology, Keio University School of Medicine, Tokyo, Japan.
  • 4. School of Life Science and Technology, Tokyo Institute of Technology, Yokohama, Japan.
  • 5. Division of Animal Sciences, Institute of Agrobiological Sciences, National Agriculture and Food Research Organization, Tsukuba, Japan.
  • 6. Laboratory of Animal Reproduction, Department of Animal Science and Resources, College of Bioresource Sciences, Nihon University, Fujisawa, Japan.
  • 7. Graduate School of Engineering, The University of Tokyo, Tokyo, Japan.
  • 8. Tamagawa Seiki Co. Ltd., Nagano, Japan.
  • 9. Department of Breast Surgery, Nippon Medical School, Tokyo, Japan.
  • 10. Department of Diagnostic Pathology, Nippon Medical School Hospital, Tokyo, Japan.
  • 11. Division of Breast Surgical Oncology, Showa University, Tokyo, Japan.
  • 12. Department of Surgery, Teikyo University School of Medicine, Tokyo, Japan.
  • 13. Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, Japan.
  • 14. Matrix Cell Research Institute Inc., Ushiku, Japan.
  • 15. Department of Nanoparticle Translational Research, Tokyo Medical University, Tokyo, Japan.
  • 16. Department of Diagnostic Pathology, Keio University Hospital, Tokyo, Japan.
PMID: 30958084 DOI: 10.1369/0022155419841023
Abstract

Current immunohistochemistry methods for diagnosing abnormal cells, such as Cancer cells, require multiple steps and can be relatively slow compared with intraoperative frozen hematoxylin and eosin staining, and are therefore rarely used for intraoperative examination. Thus, there is a need for novel rapid detection methods. We previously demonstrated that functionalized fluorescent ferrite beads (FF beads) magnetically promoted rapid immunoreactions. The aim of this study was to improve the magnetically promoted rapid immunoreaction method using antibody-coated FF beads and a magnet subjected to a magnetic field. Using frozen sections of xenograft samples of A431 human epidermoid Cancer cells that express high levels of epidermal growth factor receptor (EGFR) and anti-EGFR antibody-coated FF beads, we reduced the magnetically promoted immunohistochemistry procedure to a 1-min reaction and 1-min wash. We also determined the optimum magnetic force for the antibody reaction (from 7.79 × 10-15 N to 3.35 × 10-15 N) and washing (4.78 × 10-16 N), which are important steps in this technique. Furthermore, we stained paraffin-embedded tissue arrays and frozen sections of Metastatic Breast Cancer lymph nodes with anti-pan-cytokeratin antibody-coated FF beads to validate the utility of this system in clinical specimens. Under optimal conditions, this ultra-rapid immunostaining method may provide an ancillary method for pathological diagnosis during surgery. (J Histochem Cytochem 58:XXX-XXX, 2010).

Keywords
cancer; fluorescent ferrite beads; immunostaining; intraoperative diagnosis; magnetic field.