Cell dyn 3700

Rats were anaesthetized with ketamine-xylazine at the end of the experiment, and blood samples were collected via cardiac puncture for hematology and serum biochemistry analyses. Blood serum for biochemistry analysis was collected in non-heparinized tubes. Collected blood was left at room temperature for a while to allow clotting before serum collection by centrifugation (3000× g for 10 min) at 4 °C. The collected serum was stored at −4 °C before analysis using an automatic clinical chemistry analyzer (Hitachi, Japan). The parameters analyzed for serum biochemistry analysis were as follows: albumin, aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, creatinine, total bilirubin, urea, and total protein.
Blood was collected in K2EDTA tubes using an automated hematology analyzer (CELL-- DYN 3700 Abbott Diagnostics, Des Plaines, IL, USA). Parameters analyzed for whole blood cells were as follows: red blood cell count, hemoglobin, packed cell volume, mean corpuscular volume, mean corpuscular hemoglobin concentration, thrombocytes, and white blood cell count (including lymphocytes, monocytes, neutrophils, and eosinophils) [24 (link)].

For blood analytics, two samples were taken, namely one in a 3-mL tube with ethylenediaminetetraacetic acid (EDTA) and another in a 3.5-mL tube with polyethene terephthalate (PET). Red blood cell count was carried out in an automated Cell-Dyn 3700 analyser (Abbott Diagnostics, Chicago, IL, USA) using internal (Cell-Dyn 22) and external (Program of Excellence for Medical Laboratories-PEML) controls. Values of erythrocytes, haemoglobin, haematocrit, and haematimetry indexes were determined. These data were used to verify the health status of the subjects and were not included in the study.

Venous blood samples were taken for general analytics, in one tube of 3 mL ethylenediaminetetraacetic acid (EDTA) for hemogram, and in another tube of 3.5 mL with polyethene terephthalate (PET) for biochemical parameters. Red blood cells count (RBC) was carried out in an automated Cell-Dyn 3700 analyzer (Abbott Diagnostics, Lake Forest IL, USA), using internal (Cell-Dyn 22, Abbott Diagnostics, IL, USA) and external (Program of Excellence for Medical Laboratories-PEML) controls. Values of erythrocytes, hemoglobin, haematocrit, and hematimetric indexes (mean cell volume, MCV; mean cell haemoglobin, MCH; mean corpuscular hemoglobin concentration, MCHC; and red cell distribution width, RDW) were estimated.
Additionally, venous blood samples were collected pre VT1 test, post repeated sprint test, post second VT1 test, and 24 h after the end of the testing session for the measurement of antioxidant parameters (Figure 2). At each of the extraction points, 6 tubes of 3 mL of EDTA were obtained and one of them was centrifuged at 3500 rpm at 4 °C during 10 min and sent to the laboratory for later analysis. Urine samples, corresponding to 24 h urine collection from each participant after the supplementation, were frozen in liquid nitrogen after collection and thawed for its analysis.

Neonatal data, such as gender, neonatal weight, APGAR score, causes of admission to NICU, duration of admission in NICU, types of respiratory support used, CBC measurements (done by Cell Dyn 3700, automated cell counter, Abbott Diagnostics, USA), and thrombocytopenic manifestations, such as purpura, ecchymosis, gastric bleeding, bleeding from puncture site, and pulmonary hemorrhage or IVH, were recorded. Detailed systemic examinations focusing on skin examination, macrosomia, head circumference, intrauterine growth retardation, congenital anomalies, and dysmorphic features were recorded. Septic work-ups and blood cultures/sensitivities were performed for all included cases. A TORCH screen was carried out (by ARCHITECT i1000SR, Abbott Diagnostics, USA) in cases of suspected congenital infection. Chromosomal assays [13 , 14 (link)] were performed in cases of suspected chromosomal abnormities. Neonates suspected of having NAT, based on the presentation and clinic course of the illness, were designated as having an idiopathic cause as the diagnostic test for NAT is not available in our laboratory. Thrombocytopenia related morbidity were recorded in terms of pulmonary/IVH and mortality (alive or dead) for the study group.

The erythrogram (automated Cell-Dyn 3700, Abbott Diagnostics, Abbott Park, IL, USA) and biochemical tests (automated analyzer Architect C 8000, Abbott Diagnostics) were carried out in the Clinical Analysis Laboratory of the Clinical Hospital of the Federal University of Uberlândia. The osmotic fragility test used to evaluate the stability of erythrocytes was conducted at the Laboratory of Biophysical Chemistry of the Federal University of Uberlândia.
The analyzed hematological variables and their reference values were: erythrocytes (RBC), 4.3–5.7 millions/mm³; hemoglobin (Hb), 13.0–17.5%; hematocrit (Ht), 39–50%; mean corpuscular volume (MCV), 81.0–95.1 fL; mean corpuscular hemoglobin (MCH), 26–34 pg; mean corpuscular hemoglobin concentration (MCHC), 31–36 g/dL; red-cell distribution width (RDW), 12–15%; leucocytes (Leu), 3.5–10.5 mil/mm³; platelets (Plt), 150–450 mil/mm³; uric acid (UA), 3.5–7.2 mg/dL; creatine kinase (CK), 30–200 U/L; lactate dehydrogenase (LDH), 100–190 U/L; serum iron (Fe), 50–160 μg/dL; total cholesterol (t-C), <170 (optimum) and ≥240 mg/dL(high); high density lipoprotein cholesterol (HDL-C), ≥40 mg/dL (ideal); low density lipoprotein cholesterol (LDL-C), <130 (optimum) and≥160 mg/Dl (high); very low density lipoprotein cholesterol (VLDL-C), <40 mg/dL; and triglycerides (TGC), <150 (optimum) and ≥200 mg/dL (high).