Understanding of autophagy utilizing plasmon imaging and subcellular magnetic separation techniques

2016 Fiscal Year Final Research Report

• Project/Area Number: 26600053
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Nanobioscience
• Research Institution: Japan Advanced Institute of Science and Technology
• Principal Investigator: MAENOSONO SHINYA 北陸先端科学技術大学院大学, 先端科学技術研究科, 教授 (00323535)
• Research Collaborator: TAGUCHI TOMOHIKO 東京大学, 大学院薬学系研究科, 准教授 ; MUKAI KOJIRO 東京大学, 大学院薬学系研究科, 助教 ; TAKAKURA MASAHIRO 金沢大学, 附属病院周産母子センター, 准教授 ; WAGURI SATOSHI 福島県立医科大学, 解剖/組織学講座, 教授 ; MATSUMURA KAZUAKI 北陸先端科学技術大学院大学, 先端科学技術研究科, 准教授
• Project Period (FY): 2014-04-01 – 2017-03-31
• Keywords: オートファジー / 磁気分離 / バイオイメージング / 細胞小器官

Outline of Final Research Achievements

We developed ultrasmall magnetic-plasmonic hybrid nanobeads and applied them to the isolation of autophagosomes by applying a magnetic field. The beads were chemically synthesized and comprised an Ag/FeCo/Ag core/shell/shell structure with a mean diameter of 15 nm. The Ag core and the FeCo shell conferred imaging and magnetic separation capabilities, respectively. The nanobeads were transfected into mammalian cells by lipofection. Thirty minutes after lipofection, the nanobeads co-localized with Vps26, and subsequently with LC3. Cell lysates were prepared at the appropriate time points and were subjected to magnetic separation. The separated fraction contained LC3-II, transferrin receptor, and LAMP2, but not LC3-I, suggesting that autophagosomes of endosomal origin had been isolated.

Free Research Field

ナノ材料化学