Intravital Microscopy of Leukocyte-endothelial and Platelet-leukocyte Interactions in Mesenterial Veins in Mice.
Intravital microscopy is a method that can be used to investigate different processes in different regions and vessels in living animals. In this protocol, we describe intravital microscopy of mesentery veins. This can be performed in a short period of time with reproducible results showing leukocyte-endothelial interactions in vivo. We describe an inflammatory setting after LPS challenge of the endothelium. But in this model one can apply many different types of inflammatory conditions, like bacterial, chemical or biological and investigate the administration of drugs and their direct effects on the living animal and its impact on leukocyte recruitment. This protocol has been applied successfully to a number of different treatments of mice and their effects on inflammatory response in vessels. Herein, we describe the visualization of leukocytes and platelets by fluorescently labeling these with rhodamine 6G. Additionally, any specific imaging can be performed using targeted fluorescently labeled molecules.
Department of Cardiology and Angiology I, Heart Center, University of Freiburg.
Nadine
N
Herr
Department of Cardiology and Angiology I, Heart Center, University of Freiburg; Faculty of Biology, University of Freiburg.
Maximilian
M
Mauler
Department of Cardiology and Angiology I, Heart Center, University of Freiburg.
Christoph
C
Bode
Department of Cardiology and Angiology I, Heart Center, University of Freiburg; daniel.duerschmied@universitaets-herzzentrum.de.
Daniel
D
Duerschmied
D000818:Animals; D001792:Blood Platelets / Q000166:cytology* / Q000187:drug effects; D002450:Cell Communication / Q000187:drug effects / Q000502:physiology*; D042783:Endothelial Cells / Q000166:cytology* / Q000187:drug effects; D005456:Fluorescent Dyes / Q000737:chemistry; D007962:Leukocytes / Q000166:cytology* / Q000187:drug effects; D008070:Lipopolysaccharides / Q000494:pharmacology; D008297:Male; D008642:Mesenteric Veins / Q000166:cytology* / Q000187:drug effects; D051379:Mice; D008853:Microscopy / Q000379:methods*; D012235:Rhodamines / Q000737:chemistry
D005456:Fluorescent Dyes; D008070:Lipopolysaccharides; D012235:Rhodamines; C026188:rhodamine 6G