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Two-photon Intravital Imaging of Leukocytes During the Immune Response in Lipopolysaccharide-treated Mouse Liver

Authors
 Sang A Park  ;  Young Ho Choe  ;  Sung Hwan Lee  ;  Young-Min Hyun 
Citation
 Journal of Visualized Experiments : JoVE, Vol.0(132) : e57191, 2018 
Journal Title
 Journal of Visualized Experiments : JoVE 
Issue Date
2018
MeSH
Animals ; Humans ; Intravital Microscopy/*methods ; Leukocytes/*drug effects ; Lipopolysaccharides/pharmacology/*therapeutic use ; Liver/*physiopathology ; Mice
Abstract
Sepsis is a type of severe infection that can cause organ failure and tissue damage. Although the mortality and morbidity rates associated with sepsis are extremely high, no direct treatment or organ-related mechanism has been examined in detail in real time. The liver is the key organ that manages toxins and infections in the human body. Herein, we aimed to perform intravital imaging of mouse liver after induction of endotoxemia in order to track the motility of immune cells, such as neutrophils and liver capsular macrophages (LCMs). Accordingly, we designed a novel surgical method for exposure of the liver with minimally invasive surgery. Mice were intraperitoneally injected with lipopolysaccharide (LPS), a common endotoxin. Using our novel surgical approach for exposure and intravital imaging of the mouse liver, we found that neutrophil recruitment in LPS-treated LysM-green fluorescent protein (GFP) mouse liver was increased compared with that in phosphate-buffered saline-treated liver. After LPS treatment, the number of neutrophils increased significantly with time. Additionally, using CX3Cr1-GFP mice, we successfully visualized liver resident macrophages called LCMs. Therefore, to investigate the efficacy of new reagents to control immune mobility in vivo, determining the motility and morphology of neutrophils and LCMs in the liver may allow us to identify therapeutic effect in organ failure and tissue damage caused by leukocytes activation in sepsis.
Full Text
https://www.jove.com/video/57191/two-photon-intravital-imaging-leukocytes-during-immune-response
DOI
10.3791/57191
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Anatomy (해부학교실) > 1. Journal Papers
Yonsei Authors
Hyun, Young-Min(현영민) ORCID logo https://orcid.org/0000-0002-0567-2039
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URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/161938
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