Skip to main content
SpringerLink
Log in

Modified rapid immunohistochemical staining for intraoperative diagnosis of malignant brain tumors

  • Original Article
  • Published:
Brain Tumor Pathology Aims and scope Submit manuscript

Abstract

Rapid immunohistochemistry (R-IHC) has been developing mainly as a support technique in the rapid diagnosis of central nervous system tumors; however, there have been problems regarding instability in specimen preparation and immunostaining. To overcome the weakness of this technology, the instability of immunostaining, we developed a modified R-IHC. This was achieved by switching to 4% paraformaldehyde as the fixative solution and utilizing a commercially available Polymer Refine Detection Kit, as a high-sensitivity kit, in place of the secondary antibodies. In this study, we tested the modified R-IHC by evaluating rapid immunostaining on new staining items in 94 brain tumor removal cases, which took place at Tokyo Women’s Medical University from 2014 to 2015. The results showed that, based on GFAP and p53 markers, the modified method obtained a higher stability in specimens than the standard rapid immunostaining method. It also achieved stainability on the same level as that of a permanent specimen. The modified method tested 86.6% (46/53) and 82.8% (24/29) in pHH3 and ATRX, respectively, in the percentage of correct classification (PCC) against the permanent specimens, and 100% (7/7) in the PCC against malignant lymphomas and gliomas that used CD20/CD3 for discrimination. We concluded that the modified R-IHC method indicated a higher stainability and PCC against the permanent specimens in comparison to the standard method in GFAP, p53, CD20/CD3, pHH3, and ATRX.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Plesec TP, Prayson RA (2007) Frozen section discrepancy in the evaluation of central nervous system tumor. Arch Pathol Lab Med 131(10):1532–1540

    PubMed  Google Scholar 

  2. Uematsu Y, Owai Y, Okita R, Tanaka Y, Itakura T (2007) The usefulness and problem of intraoperative rapid diagnosis in surgical neuropathology. Brain Tumor Pathol 24(2):47–52

    Article  PubMed  Google Scholar 

  3. Louis DN, Perry A, Reifenberger G, von Deimling A, Figarella-Branger D, Cavenee WK, Ohgaki H, Wiestler OD, Kleihues P, Ellison DW (2016) The 2016 World Health Organization classification of tumors of the central nervous system: a summary. Acta Neuropathol 131(6):803–820

    Article  PubMed  Google Scholar 

  4. Masui K, Mischel PS, Reifenberger G (2016) Molecular classification of gliomas. Handb Clin Neurol 134:97–120

    Article  PubMed  Google Scholar 

  5. Kannan K, Inagaki A, Silber J, Gorovets D, Zhang J, Kastenhuber ER, Heguy A, Petrini JH, Chan TA, Huse JT (2012) Whole-exome sequencing identifies ATRX mutation as a key molecular determinant in lower-grade glioma. Oncotarget 3(10):1194–1203

    Article  PubMed  PubMed Central  Google Scholar 

  6. Arita H, Narita Y, Fukushima S, Tateishi K, Matsushita Y, Yoshida A, Miyakita Y, Ohno M, Collins VP, Kawahara N, Shibui S, Ichimura K (2013) Upregulating mutations in the TERT promoter commonly occur in adult malignant gliomas and are strongly associated with total 1p19q loss. Acta Neuropathol 126(2):267–276

    Article  CAS  PubMed  Google Scholar 

  7. Hartmann C, Hentschel B, Wick W, Capper D, Felsberg J, Simon M, Westphal M, Schackert G, Meyermann R, Pietsch T, Reifenberger G, Weller M, Loeffler M, von Deimling A (2010) Patients with IDH1 wild type anaplastic astrocytomas exhibit worse prognosis than IDH1-mutated glioblastomas, and IDH1 mutation status accounts for the unfavorable prognostic effect of higher age: implications for classification of gliomas. Acta Neuropathol 120(6):707–718

    Article  PubMed  Google Scholar 

  8. Kurimoto M, Suzuki H, Aoki K, Ohka F, Kondo G, Motomura K, Iijima K, Yamamichi A, Ranjit M, Wakabayashi T, Kimura S, Natsume A (2016) Rapid sensitive analysis of IDH1 mutation in lower-grade gliomas by automated genetic typing involving a quenching probe. Cancer Invest 34(1):12–15

    Article  CAS  PubMed  Google Scholar 

  9. Haapasalo J, Mennander A, Helen P, Haapasalo H, Isola J (2005) Ultrarapid Ki-67 immunostaining in frozen section interpretation of gliomas. J Clin Pathol 58(3):263–268

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Denda T, Kamoshida S, Kawamura J, Harada K, Kawai K, Kuwao S, Sawabe M (2013) Rapid immunocytochemistry with simple heat-induced antigen retrieval technique for improvement in the quality of cytological diagnosis. J Histochem Cytochem 61(12):920–930

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Hatta H, Tsuneyama K, Kondo T, Takano Y (2010) Development of an ultrasound-emitting device for performing rapid immunostaining procedures. J Histochem Cytochem 58(5):421–428

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Kanamori M, Kikuchi A, Watanabe M, Shibahara I, Saito R, Yamashita Y, Sonoda Y, Kumabe T, Kure S, Tominaga T (2014) Rapid and sensitive intraoperative detection of mutations in the isocitrate dehydrogenase 1 and 2 genes during surgery for glioma. J Neurosurg 120(6):1288–1297

    Article  CAS  PubMed  Google Scholar 

  13. Shioyama T, Muragaki Y, Maruyama T, Komori T, Iseki H (2013) Intraoperative flow cytometry analysis of glioma tissue for rapid determination of tumor presence and its histopathological grade: clinical article. J Neurosurg 118(6):1232–1238

    Article  PubMed  Google Scholar 

  14. Rapid Immunohistochemistry Study Group (2012) Rapid Immunohistochemistry Study Group, Japan; c2017. Available from: http://www.rihc.jp/. Accessed 1 Dec 2016

  15. Tanino M, Sasajima T, Nanjo H, Akesaka S, Kagaya M, Kimura T, Ishida Y, Oda M, Takahashi M, Sugawara T, Yoshioka T, Nishihara H, Akagami Y, Goto A, Minamiya Y, Tanaka S, R-IHC Study Group (2015) Rapid immunohistochemistry based on alternating current electric field for intraoperative diagnosis of brain tumors. Brain Tumor Pathol 32(1):12–19

    Article  CAS  PubMed  Google Scholar 

  16. Louis DN, Ohgaki H, Wiestler OD, Cavenee WK, Burger PC, Jouvet A, Scheithauer BW, Kleihues P (2007) The 2007 WHO classification of tumours of the central nervous system. Acta Neuropathol 111:97–109

    Article  Google Scholar 

  17. Terata K, Saito H, Nanjo H et al (2017) Novel rapid-immunohistochemistry using an alternating current electric field for intraoperative diagnosis of sentinel lymph nodes in breast cancer. Sci Rep 7:2810

    Article  PubMed  PubMed Central  Google Scholar 

  18. Toda H, Minamiya Y, Kagaya M et al (2011) A novel immunohistochemical staining method allows ultrarapid detection of lymph node micrometastases while conserving antibody. Acta Histochem Cytochem 44(3):133–139

    Article  PubMed  PubMed Central  Google Scholar 

  19. Varughese RK, Lind-Landström T, Habberstad AH, Salvesen Ø, Haug CS, Sundstrøm S, Torp SH (2016) Mitosin and pHH3 predict poorer survival in astrocytomas WHO grades II and III. J Clin Pathol 69(1):26–34

    Article  CAS  PubMed  Google Scholar 

  20. Wiestler B, Capper D, Holland-Letz T, Korshunov A, von Deimling A, Pfister SM, Platten M, Weller M, Wick W (2013) ATRX loss refines the classification of anaplastic gliomas and identifies a subgroup of IDH mutant astrocytic tumors with better prognosis. Acta Neuropathol 126(3):443–451

    Article  CAS  PubMed  Google Scholar 

  21. Zhang J, Wu G, Miller CP, Tatevossian RG, Dalton JD, Baker SJ, Ellison DW et al; St. Jude Children’s Research Hospital–Washington University Pediatric Cancer Genome Project (2013) Whole-genome sequencing identifies genetic alterations in pediatric low-grade gliomas. Nat Genet 45(6):602–612

  22. Tabouret E, Nguyen AT, Dehais C, Carpentier C, Ducray F, Idbaih A, Mokhtari K, Jouvet A, Uro-Coste E, Colin C, Chinot O, Loiseau H, Moyal E, Maurage CA, Polivka M, Lechapt-Zalcman E, Desenclos C, Meyronet D, Delattre JY, Figarella-Branger D, Network For POLA (2016) Prognostic impact of the 2016 WHO classification of diffuse gliomas in the French POLA cohort. Acta Neuropathol 132(4):625–634

    Article  PubMed  Google Scholar 

  23. Zhang C, Moore LM, Li X, Yung WK, Zhang W (2013) IDH1/2 mutations target a key hallmark of cancer by deregulating cellular metabolism in glioma. Neurooncology 15(9):1114–1126

    CAS  Google Scholar 

Download references

Acknowledgements

The authors are grateful to Prof. Ken Masamune, Mr. Takashi Sakayori, Dr. Manabu Tamura, and all staff members of the Department of Neurosurgery and Faculty of Advanced Techno-Surgery at Tokyo Women’s Medical University, as well as all staff members of the Ogino Memorial Laboratory at Nihon Kohden Corporation for their assistance during this study.

Author information

Authors and Affiliations

  1. Faculty of Advanced Techno-Surgery, Tokyo Women’s Medical University, 8-1 Kawada-Cho, Shinjuku-Ku, Tokyo, 162-8666, Japan

    Akane Suzuki, Takashi Maruyama, Soko Ikuta, Takakazu Kawamata & Yoshihiro Muragaki

  2. Department of Neurosurgery, Tokyo Women’s Medical University, 8-1 Kawada-Cho, Shinjuku-Ku, Tokyo, 162-8666, Japan

    Takashi Maruyama, Masayuki Nitta, Takakazu Kawamata & Yoshihiro Muragaki

  3. Nihon Kohden Corporation, Tokyo, Japan

    Akane Suzuki

  4. Tokyo Metropolitan Neurosurgical Hospital, Tokyo, Japan

    Takashi Komori

  5. Department of Surgical Pathology, Tokyo Women’s Medical University, Tokyo, Japan

    Takashi Komori

Authors
  1. Akane Suzuki
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Takashi Maruyama
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Masayuki Nitta
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Takashi Komori
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Soko Ikuta
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Takakazu Kawamata
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Yoshihiro Muragaki
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Yoshihiro Muragaki.

Ethics declarations

Conflict of interest

The authors have no conflict of interest to declare.

Ethical standards

Written informed consent was obtained from all individual patients included in this study. Consent for the publication of the case report and any additional related information was taken from the patient involved in this study.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Suzuki, A., Maruyama, T., Nitta, M. et al. Modified rapid immunohistochemical staining for intraoperative diagnosis of malignant brain tumors. Brain Tumor Pathol 34, 141–148 (2017). https://doi.org/10.1007/s10014-017-0293-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10014-017-0293-9

Keywords

Search

Navigation