成果報告書詳細
管理番号20110000001120
タイトル*平成22年度中間年報 「がん超早期診断・治療機器の総合研究開発/超早期高精度診断システムの研究開発:病理画像等認識技術の研究開発 病理画像等認識基礎技術の研究開発(定量的病理診断を可能とする病理画像認識技術)」(学校法人慶應義塾)
公開日2011/11/23
報告書年度2010 - 2010
委託先名学校法人慶應義塾
プロジェクト番号P10003
部署名バイオテクノロジー・医療技術部
和文要約和文要約等以下本編抜粋:1. 研究開発の内容及び成果等 1) 肝線維化定量法開発 肝細胞がん発生の危険因子である肝線維化を定量するために、膠原線維と弾性線維を染め分けるエラスチカ・ワンギーソン(EVG) 染色切片から、肝組織における線維化部分の占有率を算定する画像解析手法の開発に着手した(図1)。 2) 免疫蛍光デジタルスライド開発 がんの分子標的治療においては、がん組織における分子発現の適切な評価が必要である。現在、組織切片上の分子発現評価には、酵素抗体法を用いた免疫組織化学(immunohistochemistry: IHC)による評価がなされているが、その判定には議論すべき部分が多い。本来、タンパク質の定量においては、蛍光色素の方がより優れているとされているが、未だ応用はされていない。蛍光抗体法による標本は、保存性が悪く、また、組織の形態情報に乏しいため、病理診断への応用は限られたものになっている。
英文要約Title: Research and Development Project for pathological image recognition technology, Research and Development Project to develop basic technologies for recognizing pathology images, Pathological image analysis technology to enable a quantitative pathological diagnosis, (FY2010-FY2012) FY2010 Annual Report: 1. Development of digital image technology to histolgogical quantification of liver fibrosis: We started to develop the digital image technology to quantify the fibrotic area in elastica van Gieson stained sections, which discriminated collagen and elastic fibers from the other tissue components. Hepatic fibrosis is considered a risk factor for a hepatocellular carcinoma. 2. Research and development of immunofluorescent digital slide technology to quantify protein expression in archival paraffin-embedded tissue sections: Molecular targeted therapies require an adequate assessment of molecular expression in cancer tissues, but the quantification of signal intensity in immunohistochemistry (IHC) is still controversial. Immunofluorescent staining is a better method to quantify protein expression than IHC. The advanced technology of virtual slides permits digitizing a whole slide image of immunofluorescence for a few minutes. We have developed fluorescence-based, immunofluorescent quantification digital slides (IQD), a method widely applicable in routine practice. The IQD showed images of immunofluorescent and hematoxylin-stained sections simultaneously with “synchronized” or “merged” viewing. We adopted red quantum dots, which were brighter and had a much longer lifetime than organic fluorescent dyes, and its emission peak fell outside the tissue autofluorescence spectrum. After staining, bright and fluorescent imaging of whole slides were obtained with the NanoZoomer (Hamamatsu Photonics). To scan the emission of the red quantum dots, the IR cut filter was removed from an optical path of the fluorescence. IQD were established by merging the images of immunofluorescence and hematoxylin staining on exactly the same section. Initially we captured the immunofluorescent image, then stained with hematoxylin, and re-captured the image (the “two-step-scan” method). To skip the first scanning step, we attempted to scan the slides stained with hematoxylin, followed by immunostaining and capturing (the “one-step-scan” method). The membrane and cytoplasmic antigens signal was not significantly disturbed by the hematoxylin, and the localization and intensity of the immunofluorescent signal was almost the same as that with IHC. On the other hand, signals of nuclear antigens were significantly diminished, probably because the hematoxylin absorbed red emission from the quantum dots.
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