An In Vivo Model to Study Shear-Dependent Bacterial Adhesion

Abstract

Source: Claes, J., et al, In Vitro and In Vivo Model to Study Bacterial Adhesion to the Vessel Wall Under Flow Conditions. J. Vis. Exp. (2015).

This video demonstrates an in vivo method for analyzing shear-dependent bacterial adhesion in a live mouse model. Following surgical preparation and catheterization, the mesenteric vasculature is exposed for real-time imaging under a fluorescence microscope. Perfusion with an ionophore and fluorescently labeled bacteria reveals how shear stress triggers von Willebrand factor (VWF) strand formation and bacterial aggregation on endothelial surfaces.

Protocol

1. In Vivo Mesenteric Perfusion Model
   1. Preparation/surgery of the mouse
      1. Fast the mouse the night before the experiment in order to limit bowel movement.
      2. Give a 6-8 week old mouse (C57Bl/6) pre-operative analgesia by a subcutaneous injection of buprenorphine (0.1 mg/kg body weight (BW)) 20-30 min prior to the surgery.
      3. Anesthetize the mouse by intraperitoneal injection of ketamine (125 mg/kg BW) and xylazine (12.5 mg/kg BW). Check by pedal reflex. Apply vet ointment to prevent dryness.
      4. Place the mo....

Results

Figure 1. A schematic overview of the in vivo mesenteric perfusion model. A right jugular vein catheter (yellow line) is inserted for the administration of fluorescently labeled bacteria (orange dots), additional anesthetics, or other components such a.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Temgesic	Reckitt Benckiser	283716	bruprenorphine
Anesketin (Ketamin hydrochloride 115 mg/ml (100 mg/ml ketaminum))	Eurovet	BE-V136516	ketamin
XYL-M 2% (xylazine hydrochloride 23.32 mg/ml (20 mg/ml xylazine))	VMD Arendonk	BE-V170581	xylazine
2 French intravenous catheter green	Portex	200/300/010
0,9% Sodium chloride (NaCl)	Baxter Healthcare	W7124
Cotton swabs	International Medical Product	300230
Ca2+-ionophore solution A23187	Sigma-Aldrich	C7522-10 MG
26 Gauge 1 ml syringe	BD Plastipak	300013
26 Gauge 1 ml syringe with needle	BD Plastipak	300015	intra-peritoneal injection
Centrifuge 5810-R	Eppendorf	5811 000.320
Glass cover slips (24x50)	VWR	BB02405A11	Thickness No, 1
PHD 2000 Infusion	Harvard Apparatus	702100	High-accuracy Harvard infusion pump
Axio-observer DI	Carl-Zeiss	Inverted fluorescence microscope
ImageJ	National Institute of Health	Analysis software
Graphpad Prism 5,0	Graphpad Software	Analysis software
AxioCam MRm	Carl-Zeiss	Black and white camera