Dxh 800 hematology analyzer

Two reference systems were used to verify the techniques used in this study. First, a Coulter counter (DxH 800 Hematology Analyzer, Beckman Coulter, Indianapolis, IN, USA) was used to obtain the reference CBC data for the blood samples used in the analysis. Personal identity data such as names and addresses are protected by law and information regarding the individuals’ health and financial circumstances are protected. This CBC result was used as comparative data to verify the accuracy of the parameters developed for shadow image analysis. Flow cytometry (Guava easyCyte 8-10HT, Millipore, Burlington, MA, USA) was used to determine the purity of the isolated leukocyte samples. Since flow cytometry can distinguish types of blood cells using fluorescent antibodies, it is possible to confirm the purity of the separated leukocytes. The results of this analysis confirmed that high purity and high concentration leukocyte samples were obtained.

Non-fasting blood samples of 10 mL peripheral venous blood were obtained from each child by a trained pediatrician. Blood was drawn into plain and EDTA-containing VACUETTE^® Blood Collection Tubes (Greiner Bio one GmbH, Kremsmünster, Austria) handled, and stored by trained staff according to accepted protocols. The blood samples were transferred in cool conditions to a central clinical diagnostic laboratory at Hillel Yaffe Medical Center within 2 h of collection. All assays were performed on the same day in a blinded manner to the demographic, clinical, and nutritional data. A complete blood count (CBC) was conducted using a DxH-800 hematology analyzer (Beckman Coulter Inc., Brea, CA, USA). Levels of biochemical markers in serum were measured using Cobas-8000 instrument with Roche Diagnostics kits according to the manufacturer’s procedures (Roche Diagnostics GmbH, Mannheim, Germany).

Before treatment, we collected 3ml of peripheral venous blood from all patients. Serum CEA, CA125, and CA153 were measured using an automatic chemiluminescence immunoassay system (SIEMENS ADVIA centaur; Siemens, Germany). NLR was the absolute neutrophil count/the total lymphocyte, LMR was the total lymphocyte/the absolute monocyte, and PLR was the total platelet count/the total lymphocyte count. These parameters were analyzed from the peripheral blood cell count (DxH 800 hematology analyzer, Beckman Coulter). See Supplementary for the experimental methods of cellular immunology related indicators.

Peripheral venous blood samples were sampled via the antecubital vein in the morning after an overnight fasting. Complete blood counts and differential counts were analyzed by the automated cellular analysis system Beckman Coulter DxH 800 hematology analyzer (Beckman Coulter, Miami, FL). Biochemical parameters of albumin, lipid profile, high sensitive-C reactive protein (hs-CRP), fasting glucose, aspartate transaminase, blood urea nitrogen, and serum creatinine were measured by the ARCHITECT i2000SR immunoassay analyzer (Abbott Diagnostics, Abbott Park, IL, USA). Lymphopenia was defined by absolute lymphocyte count < 1000/cumm [29 (link)].
Lymphocyte subsets were determined flow-cytometric analysis system Beckman Coulter FC500 flow cytometer (Beckman Coulter, Miami, FL) in freshly drawn peripheral blood after adequate processing as the manufacturer’s instruction. Serum CMV IgG, VZV IgG, HBsAg IgG, HCV IgG, and C. pneumoniae IgG were measured by a chemiluminescent immunoassay on the ARCHITECT i2000SR immunoassay analyzer (Abbott Diagnostics, Abbott Park, IL, USA).

All CBC analyses were performed with an automatic hematologic analyzer (Beckmancoulter DxH 800 Hematology Analyzer). Hemoglobin (Hb), white blood cell (WBC), neutrophils, lymphocytes, monocytes, eosinophils, and platelet counts were obtained before surgical treatment. Blood samples were collected in di-potassium ethylenediaminetetraacetic acid tubes.

All biochemical procedures were done on the first day of hospital admission in a specialized biochemical laboratory of the Clinical Center Kragujevac, Serbia. Complete blood cell count (CBC) was measured using a hematology analyzer (DxH 800 Hematology Analyzer by Beckman Coulter). The biochemical parameters such as glucose, creatinine, urea, cholesterol, triglyceride (TG), aspartate aminotransferase (AST), alanine aminotransferase (ALT) gamma-glutamyl transferase (GGT), lactate dehydrogenase (LDH), total and direct bilirubin, C-reactive protein (CRP), sodium, and potassium were estimated from the serum samples by using standard kits in an automatic clinical chemistry analyzer (AU680 Clinical Chemistry Analyzer by Beckman Coulter). Measurement of the vitamin D level was performed using an automated immunoassay analyzer—the Alinity i system (Abbott Laboratories, IL, USA) that utilizes the chemiluminescent microparticle immunoassay (CMIA) principle. The level of procalcitonin in the serum was determined by the method of electrochemiluminescence, on the immunochemistry analyzer (Cobas e 411 by Roche). D dimer concentration measurement was performed on coagulation analyzer ACL-TOP 300 (Instrumentation Laboratory, Bedford, USA) employing the automated latex-enhanced particle immunoturbidimetric method.