Complete blood counts were obtained by Beckman Coulter Hematology analyzer (Brea, CA, United States). Peripheral blood mononuclear cells (PBMC) and plasma were obtained by standard density gradient centrifugation over the blood separation polymer Ficoll (GE Healthcare Life Sciences, Pittsburg, PA, United States). PBMC were frozen in 10% DMSO/FBS and stored in liquid nitrogen while plasma was stored at -80°C until analysis.
Hematology analyzer
Manufactured by Beckman Coulter
Sourced in United States
The Hematology Analyzer is a diagnostic instrument used to analyze and count various blood cells, including red blood cells, white blood cells, and platelets. It provides accurate and reliable results for a comprehensive assessment of a patient's hematological status.
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Lab products found in correlation
Ficoll, by GE Healthcare (5 mentions)
Immunocap system, by Thermo Fisher Scientific (3 mentions)
High performance liquid chromatography (hplc), by Bio-Rad (1 mentions)
Gallios flow cytometer, by Beckman Coulter (1 mentions)
Integra 400, by Roche (1 mentions)
7150 analyzer, by Hitachi (1 mentions)
Cobas 6000 c501, by Roche (1 mentions)
Idk calprotectin, by Immundiagnostik (1 mentions)
Cobas e411, by Roche (1 mentions)
Mr frosty freezing container, by Thermo Fisher Scientific (1 mentions)
Enzyme immunoassay kit, by Bachem (1 mentions)
Immulite 2000, by Siemens (1 mentions)
Genomic dna purification kit, by Promega (1 mentions)
Enzyme linked immunosorbent assay kit, by Cusabio (1 mentions)
Coat a count, by Siemens (1 mentions)
Cobas c501, by Roche (1 mentions)
Cytometric bead array, by BD (1 mentions)
30 protocols using hematology analyzer
1
Isolation and Cryopreservation of PBMC
2
Chlamydomonas reinhardtii Cell Density Analysis
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In terms of densitometry, the cell density of Chlamydomonas reinhardtii was positively correlated with the OD750, which could be expressed via a detection of the OD750 of the cultures, after mixing 3–4 mL of the tested Chlamydomonas reinhardtii cultures and pouring the mixture into a cuvette to detect the absorbance value at 750 nm using an ultraviolet spectrophotometer.
In terms of cytometry, using dilutions supplied with the Beckman Coulter hematology analyzer, 1 mL of the tested cells was taken and diluted 20-fold (40-fold dilution in the logarithmic growth phase) for counting using the Beckman Coulter Z2 cytometer (https://www.beckmancoulter.cn/ , accessed on 8 October 2020).
In terms of cytometry, using dilutions supplied with the Beckman Coulter hematology analyzer, 1 mL of the tested cells was taken and diluted 20-fold (40-fold dilution in the logarithmic growth phase) for counting using the Beckman Coulter Z2 cytometer (
3
Sickle Cell Disease Hematological Profile
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Patients with SCD (n = 47) and healthy controls (n = 46) aged 20 to 59 years were enrolled in the present study. Participants with HbSS or HbSβ0 genotypes were included as patients with SCD, and hemoglobin patterns were confirmed by high-performance liquid chromatography (HPLC) (Bio-Rad) and DNA sequence analysis. Patients who had received blood transfusions in the past 3 months, in vaso-occlusive crises and with apparent infection were excluded from the study. No patient was being treated with antibiotics or corticosteroids, and all patients were under treatment with folic acid, calcium, and vitamin D. Moreover, most patients were under treatment with hydroxyurea. Blood samples were obtained from the participants during regular consultation at the Unicamp Hematology and Hemotherapy Center, São Paulo, Brazil. Complete blood counts with reticulocyte counts were performed on blood collected with EDTA in a hematology analyzer (Beckman Coulter). The study was approved by the Unicamp Human Research Ethics Committee (protocol number CAAE: 85061318.0.0000.5404). All patients and controls had agreed to participate and had signed informed consent forms.
4
Comprehensive Blood Cell Analysis
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A white blood cell count (WBC) and differential was performed using a Beckman-Coulter Hematology Analyzer. The peripheral immunophenotype consists of leukocyte differential, lymphocyte subsets, CD4/CD8 ratio and memory/naïve T cell subsets. Levels of cytotoxic T cells, central memory T cells, and NK cell/B cell/monocyte subsets were also assessed by multicolor flow cytometry. Cell surface markers were stained by first combining 100 µl of EDTA whole blood and 10 ug of each appropriate labeled monoclonal antibodies. Staining was performed by incubation at room temperature for 20 min. Red blood cells were lysed using Beckman-Coulter Optilyse as described by the manufacturer. Stained leukocytes were then fixed in 1.0% paraformaldehyde in PBS for 10 min and analyzed on a Beckman-Coulter Gallios flow cytometer.
5
Healthy Donor Blood Cell Analysis
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Blood samples from 10 anonymous healthy individuals were collected in ethylenediaminetetraacetic acid (EDTA) tubes after written informed consent from the Experimental Centre for Technical Medicine (ECTM) donor service (University of Twente, Enschede, The Netherlands). The frequencies of white blood cells, red blood cells, and platelets were assessed using a hematology analyzer (Beckman Coulter, Brea, CA, USA). The samples were processed on the same day of the drawing.
6
Comprehensive Metabolic Profiling Protocol
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Hemoglobin A1c testing was performed by Integra 400 (Roche Diagnostics, Laval, QC, Canada). Fasting blood glucose level was performed daily by Dextrostix (Dtx) with a Glucocheck meter. Blood chemistry tests were performed by the Siemens Advia 1800 analyzer. Complete blood count was performed by Beckman Coulter hematology analyzer.
7
Effects of Exercise on Inflammatory Markers
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Subjects’ blood samples for WBC count and serum were obtained before and immediately (1-5 minutes) following RE. WBC count was performed by the Beckman Coulter hematology analyzer (USA). Sera were isolated and stored at -20°C until use. Measurement of IL-15 serum levels was performed by an enzyme-linked immunosorbent assay (ELISA) kit (R and D Systems, Minneapolis, MN) according to the manufacturer’s protocol. The sensitivity of test was 0.2 pg/mL. TNF-α and hs-CRP levels were measured by IBL Company ELISA kit (Hamburg, Germany) as described by the manufacturer. The sensitivity of the tests was 2.3 pg/mL and 0.02 µg/mL, respectively.
8
Evaluation of Liver Disease Indicators
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All patients were evaluated on standard laboratory parameters. The complete blood count was measured on Hematology Analyzer (Beckman Coulter 5 diff, Miami FL) and clinical chemistry tests were performed using 7150 Analyzer (Hitachi, Japan). All recorded indicators were from blood routine examination, coagulation function, liver and kidney function, serum lipid, myocardial enzyme and demographic characteristics. Thirty-nine variables were excluded in this study, because of literature and medical background (28 predictors such as Chlorine, Urea, Uric acid), as well as over-missing values (11 predictors such as C-reactive protein, Hepatitis b E antigen, HBeAb, HBV-DNA).
The variable “sign” is primarily considered as an indicator of clinical feature, which represents the status and symptoms of patients. If a patient had both liver palms and spider nevus, the “sign” was assigned to 2. If a patient had either liver palms or spider nevus, the “sign” was assigned to 1. If a patient has neither liver palms nor spider nevus, the “sign” was assigned to 0.
The variable “sign” is primarily considered as an indicator of clinical feature, which represents the status and symptoms of patients. If a patient had both liver palms and spider nevus, the “sign” was assigned to 2. If a patient had either liver palms or spider nevus, the “sign” was assigned to 1. If a patient has neither liver palms nor spider nevus, the “sign” was assigned to 0.
9
Metabolic Profiling of Participants
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Demographic data was collected from all subjects (age and sex), and all subjects underwent physical examination, including measurement of height, body weight, and waist circumference. Overnight fasting blood samples were obtained to test for hemoglobin, HbA1c, liver and renal functions, and lipid profiles. An OGTT was performed to collect blood samples at 0, 30, 60, and 120 min of the OGTT to measure glucose and insulin levels. Plasma glucose, liver and renal functions, and lipid profiles were measured using a Hitachi 7180 automated analyzer (Hitachi High-Tech Science Systems Corporation, Hitachinaka-shi, Japan). Hemoglobin was measured using a Hematology analyzer (Beckman Coulter Inc, California, USA). HbA1c was measured with a high-performance liquid chromatography analyzer (Bio-Rad Labs, Brea, CA, USA). Serum insulin was measured with radioimmunoassay (Roche Diagnostics, Mannheim, Germany).
10
Blood Sample Processing Workflow
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Complete blood counts were obtained by Beckman Coulter hematology analyzer (Brea, CA). Peripheral blood mononuclear cells (PBMC) and plasma were obtained by standard density gradient centrifugation over Ficoll (GE Healthcare). PBMC were frozen in 10% DMSO/FBS and stored in liquid nitrogen until analysis. Plasma was stored at −80°C until analysis.
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