Six milliliters of whole blood was collected from each animal on inoculation day and 7 days thereafter for processing blood cell count and cytokine assays. Blood was collected from femoral vein under anesthesia attained by intramuscular injection of a ketamine-xylazine mixture with ketamine HCl at a dosage of 10 mg/kg body weight and xylazine 5%. While 3 ml of the collected blood was processed for cell count, the other 3 ml were processed for serum, which was then stored at −20 °C. Cell counting was done using an automated differential cell counter (Coulter Ac·T 5diff CP, Beckman Coulter, Miami, FL, USA). Serum was separated from whole blood through centrifugation using a Jouan C422 centrifuge (LabCare America, FL, USA) at 1500×g for 10 min and then collected into a clean tube for storage.
Coulter ac t 5diff cp
Manufactured by Beckman Coulter
Sourced in United States, France
The Coulter Ac·T 5diff CP is a hematology analyzer that provides complete blood count (CBC) and 5-part differential (5diff) analysis. It performs automated analysis of red blood cells, white blood cells, and platelets in a blood sample.
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Lab products found in correlation
3 protocols using coulter ac t 5diff cp
1
Blood Sample Collection and Processing
2
Biochemical Indicators in Child Blood
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In the original studies, blood samples collected from the children were used to analyze biochemical indicators. In the study in KwaZulu-Natal, Hb concentrations were measured on whole blood, using the direct cyanmethemoglobin method (Bio Rad Laboratories PTY Ltd.) by using Drabkin solution and a standard miniphotometer [24] , while in the two studies in North West, a hematology analyzer (Coulter Ac-T 5diff CP; Beckman Coulter) was used [25, 26] . PF and CRP were measured using an automated chemiluminescent immunoassay system (IMMULITE; Siemens) [24] in the study in KwaZulu-Natal, while in the two studies in North West, a ferritin enzyme-linked immunosorbent assay kit (Ramco Laboratories Inc.) was used for PF and an immunoturbidimetric method (Technicon RAÀ1000 auto analyzer; Bayer) for CRP [25, 26] . For the pooled-data analysis, the PF values of children with a CRP concentration 5 mg/L were adjusted for inflammation by multiplying PF values by a correction factor of 0.65 [27] . Anemia was defined as Hb < 115 g/L, ID as PF < 15 mg/L, and ID anemia as a combination of Hb < 115 g/L and PF < 15 mg/L [5, 28] .
3
Blood Biochemistry Analysis Protocol
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A portion of each blood sample was introduced in situ into a blood glucose analyzer (Accu-chek, Roche, Spain) to determine blood glucose concentration. The whole blood was also analyzed for hemoglobin concentration (Coulter ACT5 Diff CP; Beckman-Coulter Instruments, Villepinte, France) and hematocrit was measured by triplicate using microcentrifugation. Afterwards, the blood samples were allowed to clot and they were centrifuged at 5000 g for 10 min to obtain serum samples. The serum samples were introduced in clean tubes and frozen at -80°C until they were analyzed for creatine kinase and myoglobin concentrations by means of an autoanalyzer (Access II, Beckman-Coulter Instruments, USA) in a latter date. Serum electrolyte concentrations (Na+, Cl- and K+) were also determined in all serum samples (Spotlyte, Menarini Diagnostics, Madrid, Spain).
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