In order to investigate the rolling velocity of murine neutrophils on E- or P-selectin, we used an autoperfused flow chamber (6 (link), 31 (link), 33 (link)). Rectangular glass capillaries (20×200 µm) were filled either with E-selectin (2.5 µg/ml, R&D Systems, MN, USA) or P-selectin (20 µg/ml, R&D Systems) alone or in combination with ICAM-1 (2 µg/ml in combination with E-selectin, 5 µg/ml in combination with P-selectin, R&D Systems) for 2 h and then blocked for 2 h using casein (1%, Thermo Scientific). One side of the chamber was connected to a mouse carotid artery. The other side of the chamber was connected to a PE 50 tubing (Becton Dickinson, Sparks, MD, USA) and used to control the wall shear stress in the capillary. To investigate chemokine-induced adhesion in vitro (33 (link)), glass capillaries were coated with P-selectin (50 µg/ml) and ICAM-1 (15 µg/ml) or P-selectin and ICAM-1 in combination with CXCL1 (25 µg/ml).
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Investigating Murine Neutrophil Rolling Velocity
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Corresponding Organization : Max Planck Institute for Molecular Biomedicine
Other organizations : University of California, San Francisco
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Variable analysis
independent variables
- Selectin type (E-selectin or P-selectin)
- Presence of ICAM-1 (with or without)
- Rolling velocity of murine neutrophils
- Rectangular glass capillaries (20×200 µm)
- Coating concentrations (E-selectin: 2.5 µg/ml, P-selectin: 20 µg/ml, ICAM-1: 2 µg/ml with E-selectin, 5 µg/ml with P-selectin)
- Coating duration (2 h)
- Blocking with casein (1%, 2 h)
- Mouse carotid artery
- Wall shear stress in the capillary
- P-selectin (50 µg/ml) and ICAM-1 (15 µg/ml)
- P-selectin and ICAM-1 in combination with CXCL1 (25 µg/ml)
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