To study the effect of VN-deficiency on immune cell infiltration of liver, kidneys, and lungs of mice with severe systemic inflammation, organs were harvested 6 h after intraperitoneal injection of LPS (see above) and homogenized. The platelet count as well as the total white blood count of each organ homogenate was determined using a ProCyte Dx cell counter (IDEXX Laboratories, Westbrook, ME, USA). After incubation with anti-CD45-APC/Cy7 mAb (clone 30-F11; BD Bioscience), anti-CD11b-FITC mAb (clone M1/70; eBioscience, Waltham, Massachusetts, USA), anti-GR-1-PE mAb (clone RB6-8c5; eBioscience), anti-F4/80-eFluor 450 mAb (clone BM8; eBioscience) for 30 min on ice, lysis of erythrocytes was performed using lysing solution (BD Biosciences) for 10 min at room temperature. After washing, immunostained cells were analyzed on a flow cytometer (Gallios flow cytometer; Beckman Coulter Inc, Brea, California, USA). Approximately 20,000 gated events were collected in each analysis and isotype-matched controls were used in all experiments. Data analysis was performed using FlowJo software (Becton, Dickinson and Company). Absolute immune cell numbers were calculated using the white blood count and the relative share of the respective immune cell subset as assessed by flow cytometry.
Procyte dx cell counter
Manufactured by IDEXX
Sourced in United States
The ProCyte Dx cell counter is a laboratory instrument designed to perform automated blood cell analysis. It provides accurate and efficient counting and classification of various blood cell types, including red blood cells, white blood cells, and platelets.
Automatically generated - may contain errors
Lab products found in correlation
Lysing solution, by BD (1 mentions)
Anti cd45 apc cy7 mab clone 30 f11, by BD (1 mentions)
Anti cd11b fitc mab clone m1 70, by Thermo Fisher Scientific (1 mentions)
Anti gr 1 pe mab clone rb6 8c5, by Thermo Fisher Scientific (1 mentions)
Anti f4 80 efluor450 mab clone bm8, by Thermo Fisher Scientific (1 mentions)
Gallios flow cytometer, by Beckman Coulter (1 mentions)
2 protocols using procyte dx cell counter
1
Assessing Immune Cell Infiltration in Organs of Mice with Severe Inflammation
2
Mesenteric Lymph Vessel Cannulation
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40 min prior to cannulation, 200 μl olive oil was administered intragastrally to identify mesenteric lymph vessels that are located just upstream of the thoracic duct. Mice were anesthetized (100 mg/kg ketamine, 20 mg/kg xylazine, 1% acepromazin i.p.), vessels were cannulated and lymph was drawn via a fine bore polythene tubing (Smiths Medical) that had previously been flushed with PBS containing EDTA. In some experiments, mice were injected i.p. with 1 mg/kg (or less) of FTY720 or 10 mg/kg of SEW2871 1 or 2 hours, respectively, before cannulation. Cell numbers were determined using an IDEXX ProCyte DX cell counter.
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