Intravital imaging of microcirculation on murine sepsis model using two-photon laser scanning microscopy

2015 Fiscal Year Final Research Report

• Project/Area Number: 25462053
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Digestive surgery
• Research Institution: Mie University
• Principal Investigator: KOBAYASHI MINAKO 三重大学, 医学(系)研究科(研究院), 講師 (30359765)
• Co-Investigator(Kenkyū-buntansha): KOIKE Yuki 三重大学, 医学部附属病院, 助教 (10555551) ; TOIYAMA Yuji 三重大学, 医学系研究科, 助手 (00422824) ; TANAKA Koji 三重大学, 医学部付属病院, 講師 (10345986) ; MOHRI Yasuhiko 三重大学, 医学系研究科, 准教授 (70345974) ; MIZIGUCHI Akira 三重大学, 医学系研究科, 教授 (90181916) ; KUSUNOKI Masato 三重大学, 医学系研究科, 教授 (50192026)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Keywords: 二光子励起顕微鏡 / 敗血症 / リアルタイムイメージング

Outline of Final Research Achievements

We developed a method of intravital imaging of interaction between leukocytes, platelets and vascular endothelium in tissue microcirculation of a lipopolysaccharide -induced sepsis model using a two photon laser scanning microscopy.
We imaged the rolling or adhesion of leukocytes to vascular endothelium, leukocyte-platelet aggregates, and platelet aggregation in postcapillary venules of the cecum and hepatic sinusoids in vivo real-time. We also imaged neutrophil extracellular traps (NETs) which are activated by these interactions. NETs were classified as two distinct forms; cell-free NETs that were released away from neutrophils (fragmented or cotton-like structures) and anchored NETs that were anchored to neutrophils (linear, reticular, membranous, or spot-like structures). NETs seemed to be associated with the formation of platelet aggregates or leukocyte-platelet aggregates. These observations may suggest the adverse effect of intravascular NETs on the host during a sepsis.

Free Research Field

消化器外科