Act 5 diff hematology analyzer

PRP was extracted using a two-stage density gradient centrifugation method. About 30 mL venous blood were drawn in a sterile syringe containing 600 U heparin calcium (1 mL:10,000 U). The blood sample was centrifuged at 1200 r/min for 10 minutes. Subsequently, plasma, buffy coat, and 2–3 mm red blood cells were collected, mixed, and then centrifuged at 3500 r/min for 5 minutes. About 1/2 volume of platelet-poor plasma (PPP) was discarded, and the remaining PPP was resuspended to obtain PRP. Platelet count of all samples of whole blood and PRP were detected through A^c·T™ 5diff Hematology analyzer (Beckman Coulter, Inc., Brea, CA), and platelet concentration was 700 × 10⁹/L to 1000 × 10⁹/L. Lastly, PRP was adequately mixed with calcium gluconate at the ratio of 9:1 to activate platelets.

We collected blood samples in the morning after overnight fasting. Venous blood (10 mL) was collected from each patient, and part of it was transferred to the EDTA (ethylenediaminetetraacetic acid) coated vacutainers for the estimation of hematological parameters. The remaining blood was allowed to clot and centrifuged at 300 rpm for 10 minutes to obtain serum and stored at–20°C to measure the rest of the analytes.
Hematological analysis included the estimation of hemoglobin, total white blood cells, red blood cells, platelets, packed cell volume, mean corpuscular volume, mean corpuscular hemoglobin, and erythrocyte sedimentation rate. These were analyzed using the Ac T 5diff Hematology Analyzer (Beckman Coulter). Biochemical analysis included plasma glucose, serum creatinine blood urea, liver enzymes, and lipid profile (total cholesterol, triglycerides, high-density lipoprotein cholesterol, and very low density lipoprotein cholesterol).