Adhesion of effector CTLs was assessed as described by Matthews and colleagues43 (link). Briefly, Ibidi VI0.4 m-slides were coated with 6 mg/mL ICAM-1 overnight at 4°C. Effector CD8 T cells at a density of 106 cells/mL in adhesion medium (RPMI with 0.1% BSA, 40 mM HEPES and 2 mM MgCl2) were injected into a flow system that used a silicone tubing loop connected to a Multi-phaser NE-1000 syringe pump (New Era Pump Systems Inc.). Cells were allowed to flow over the ICAM-1-coated plates at a continuous shear flow rate of 0.5 dynes / cm2 for 10 mins. Cells were monitored by microscopy and the number of adherent cells in the field of view was determined at 1 min intervals by manual counting.
Multi phaser ne 1000 syringe pump
Manufactured by New Era Pump Systems
The Multi-phaser NE-1000 is a syringe pump designed for precision fluid delivery. It features independent control of flow rate and infusion/withdrawal volumes. The device is capable of supporting a wide range of syringe sizes.
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2 protocols using multi phaser ne 1000 syringe pump
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Effector CTL Adhesion Assay
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Effector CTL Adhesion Assay
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Adhesion of effector CTLs was assessed as described by Matthews and colleagues43 (link). Briefly, Ibidi VI0.4 m-slides were coated with 6 mg/mL ICAM-1 overnight at 4°C. Effector CD8 T cells at a density of 106 cells/mL in adhesion medium (RPMI with 0.1% BSA, 40 mM HEPES and 2 mM MgCl2) were injected into a flow system that used a silicone tubing loop connected to a Multi-phaser NE-1000 syringe pump (New Era Pump Systems Inc.). Cells were allowed to flow over the ICAM-1-coated plates at a continuous shear flow rate of 0.5 dynes / cm2 for 10 mins. Cells were monitored by microscopy and the number of adherent cells in the field of view was determined at 1 min intervals by manual counting.
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