Establishment of Electron microscopy high-resolution CLEM method utilizing FFPE specimens Exploration of new renal transplantation pathology diagnosis

2022 Fiscal Year Final Research Report

• Project/Area Number: 19K07423
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 49020:Human pathology-related
• Research Institution: Showa University
• Principal Investigator: TAKAKI TAKASHI 昭和大学, 大学共同利用機関等の部局等, 准教授 (10774820)
• Co-Investigator(Kenkyū-buntansha): 康 徳東 昭和大学, 医学部, 講師 (00571952) ; 本田 一穂 昭和大学, 医学部, 教授 (10256505)
• Project Period (FY): 2019-04-01 – 2023-03-31
• Keywords: 免疫電顕染色 / 走査電子顕微鏡 / ホルマリン固定パラフィンブロック / CLEM法 / 腎生検 / 移植腎 / 病理診断

Outline of Final Research Achievements

Using formalin-fixed paraffin-embedded (FFPE) samples, which are widely used in pathology, we established an immunostaining technique to extensively visualize minute positive areas that are difficult to visualize with optical microscopy using scanning electron microscopy in order to determine if renal transplantation is a good or bad decision. Using image analysis after staining FFPE sections after renal transplantation with this immunostaining, we analyzed and compared the correlation and initial trends with rejection after renal transplantation using five different analysis software, but no significant differences were found.
However, a patent is pending for a case combining the immunostaining technique and microscopy obtained in the course of this research and the development of a new slide that can be deparaffinized without the use of xylene.

Free Research Field

病理学

Academic Significance and Societal Importance of the Research Achievements

Ｔ細胞関連型拒絶反応、抗体関連型拒絶反応、移植後ウイルス感染と移植拒絶の移植前後の画像解析結果を、症例毎に比較検討を行ったが、画像解析の結果から有意な差は認められなかった。しかしこの研究を行う事で、FFPE標本を用いた走査電子顕微鏡の免疫染色法の確立、この染色を用いた新たな解析装置の開発、キシレンによる脱パラフィンが不要となる新たなスライドの開発とそれに関連した2件の特許を現在申請中である。