Intravital Imaging of Neutrophil Recruitment in intestinal Ischemia-Reperfusion Injury in Mice using two-photon laser scanning microscopy

2016 Fiscal Year Final Research Report

• Project/Area Number: 26461918
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: General surgery
• Research Institution: Kumamoto University
• Principal Investigator: Honda Masaki 熊本大学, 医学部附属病院, 非常勤診療医師 (80573609)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Keywords: 小腸虚血再灌流障害 / 2光子励起顕微鏡 / 好中球動態 / 生体内イメージング / 微小循環

Outline of Final Research Achievements

Activated neutrophils induce local inflammation and result in pathophysiologic changes during intestinal ischemia-reperfusion (I/R) injury. However, few studies have examined real-time intravital neutrophil recruitment. Here, we show a method for imaging and dynamic change of the neutrophil recruitment in intestinal I/R injury using two-photon laser scanning microscopy (TPLSM).
LysM-eGFP mice were subjected to 45 min of warm intestinal ischemia followed by reperfusion. Using time-lapse TPLSM technique, we directly observed the behavior of neutrophils in intestinal I/R injury. At low magnification, the number of neutrophils per field of view continued to increase for 4 hour after reperfusion. High magnification images revealed the presence or absence of blood circulation.
TPLSM imaging could reveal the real-time neutrophil recruitment in intestinal I/R injury. Z-stack imaging was useful to evaluate pathophysiological changes of intestinal wall.

Free Research Field

小児外科・移植外科・生体内イメージング