Normal haemoglobin HGB levels range from 130–175 g/L for men and 120–155 g/L for women. The Sysmex XP-300 (Sysmex Corporation, Kobe, Japan) automated haematology analyser was used to conduct HGB analysis. The Sysmex XP-300 is compliant with International Standards Organisation (ISO)/International Electrotechnical Commission (IEC) 17043:2010, via the Sysmex Network Communication Service. A total of 50 μL of each sample was aspirated by the Sysmex XP-300. A non-cyanide haemoglobin detection method [60 (link)] was used for HGB analysis. Calibration of the XP-300 was performed by Sysmex Corporation according to the manufacturer’s specifications; quality was assured using Sysmex internal quality control.
Xp 300
Manufactured by Sysmex
Sourced in Japan, Germany, United States
The XP-300 is a compact, automated hematology analyzer designed for small- to medium-sized laboratories. It performs complete blood count (CBC) analysis, including red blood cell, white blood cell, and platelet parameters.
Automatically generated - may contain errors
Lab products found in correlation
Automated hematology analyzer, by Sysmex (3 mentions)
Optiprep, by Merck Group (2 mentions)
Lsr 2 flow cytometer, by BD (2 mentions)
Facsdiva software, by BD (2 mentions)
Facscanto 2, by BD (2 mentions)
Xn 9000, by Sysmex (1 mentions)
Rpmi media, by Thermo Fisher Scientific (1 mentions)
U46619, by Bio-Techne (1 mentions)
Cellquest software, by BD (1 mentions)
Kx 21, by Sysmex (1 mentions)
Coagulation sodium citrate 3.2, by Greiner (1 mentions)
Vacutainer 7.2mg k2edta, by BD (1 mentions)
Facscanto 2 flow cytometer, by BD (1 mentions)
Modular auto analyser, by Roche (1 mentions)
Sepharose cl 2b, by GE Healthcare (1 mentions)
Ny2002500, by Merck Group (1 mentions)
Amicon ultra 2 ml, by Merck Group (1 mentions)
Sepmate tube, by STEMCELL (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Phosphate buffered saline (pbs), by Biowest (1 mentions)
81 protocols using xp 300
1
Automated Hemoglobin Analysis Using Sysmex XP-300
2
Automated Hemoglobin Analysis using Sysmex XP-300
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HGB, a secondary outcome measure, was analysed using the Sysmex XP-300 (Sysmex Corporation, Kobe, Japan) automated haematology analyser. The Sysmex XP-300 is compliant with International Standards Organisation (ISO)/International Electrotechnical Commission (IEC) 17043:2010, via the Sysmex Network Communication Service. The Sysmex instrument aspirated 50 μL of each sample. HGB was analysed using a non-cyanide haemoglobin detection method [51 (link)]. The XP-300 was calibrated by Sysmex Corporation according to the manufacturer’s specifications, and quality was guaranteed using Sysmex internal quality control.
3
Comprehensive Blood Analysis Protocol
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About 5 mL of blood was collected in Ethylene a Diamine Tetraacetic Acid (EDTA) test tube from participants who completed the questionnaire and who agreed to give blood. Four workers did not volunteer to participate, and an additional two were excluded due to acute infection and pregnancy cases. Complete blood count (CBC) tests, including total RBC count, total WBC count, Hb and Hct levels, total platelet, MCV, MCH, MCHC, mean platelet volume, and red blood cell distribution width (RDW), absolute and relative counts of lymphocyte and neutrophil, were analyzed using the 3-part hematological auto analyzer (Sysmex XP-300, Sysmex Corporation, Kobe, Japan) within 2 hours of blood collection. Sysmex XP-300 performs rapid and accurate analysis of a 17-parameter CBC, including a 3-part WBC differential.36
4
Platelet Enumeration in Whole Blood and PRP
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EDTA anticoagulated whole blood was measured on Sysmex®XN-9000 and Sysmex® XP-300 according to the instruction manual. In short, undiluted whole blood was aspirated into the devices and platelet count was obtained. Whole blood platelet count was determined with the PC100 platelet counter after manual dilution (1:25) of the whole blood with ammonium oxalate solution (ThromboCount Pur; Bioanalytic GmbH, Umkirch/Freiburg, Germany). Platelet enumeration was performed in two technical replicates by using single glass slides with two adjacent chambers (chamber A and B).
Prior to platelet count determination in PRP, the validation samples were diluted to 250 × 103/μl with homologous PPP and subsequently were measured by Sysmex® XP-300 in a single measurement. The PRP validation samples were diluted (1:100) with ThromboCount Pur and subsequently measured with PC100 platelet counters as described above for the whole blood samples.
Prior to platelet count determination in PRP, the validation samples were diluted to 250 × 103/μl with homologous PPP and subsequently were measured by Sysmex® XP-300 in a single measurement. The PRP validation samples were diluted (1:100) with ThromboCount Pur and subsequently measured with PC100 platelet counters as described above for the whole blood samples.
5
Bronchoalveolar Lavage Cell Analysis
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The trachea was exposed via blunt dissection and cannulated. A syringe was used to introduce RPMI media (Life Technologies) into the lungs for 30 s, before being withdrawn. 0.3 mls was introduced 3 times. The samples were then pooled to give 1 sample per animal, before being prepared for total and differential cell counts. The total leukocyte cell counts in BAL fluid were attained using a Sysmex XP-300 automated cell counter (Sysmex Ltd., UK). Slides for differential cell counts were prepared using 100 μl BAL fluid in a cytospin (807 g, 5 min, room temperature, low acceleration) (Shandon, Runcorn, UK). Slides were stained using a Hema-tek 2000 automated slide stainer (Ames Co., Elkhart, USA) using ACCUSTAIN® modified Wright-Giemsa stain (Sigma). Slides were analysed under light microscopy at ×40 magnification by an observer blinded to the specimen identities. Differential counts were completed on 200 cells per slide using standard morphological criteria. The percentage of macrophages/monocytes, neutrophils, eosinophils and lymphocytes were calculated. Macrophages and monocytes were counted as one group. Other cell types such as epithelial cells and red blood cells were ignored.
6
Platelet Aggregation Assay in Mice
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Platelet-rich plasma (PRP) was collected from Pf4-Cre+Gucy1b1+/LoxP and Pf4-Cre+Gucy1b1LoxP/LoxP mice as stated in the manuscript and thrombocyte count was measured using an automated hematology analyzer (XP-300; Sysmex Corp). PRP was centrifuged at 700g for 10 min to receive PPP used for blanking. Samples were incubated at 37 °C in glass cuvettes with constant stirring on an 8-channel personal computer-controlled platelet aggregation profiler (PAP-8, Biodata Corp) either in the presence of sodium nitroprusside (final concentration 10 μmol l−1, catalog no. HN34.1; Carl Roth), BAY-747 (150 ppm) or vehicle (dimethyl sulfoxide (DMSO), 0.4%) for 2 min. Subsequently, thrombocyte aggregation was induced by the addition of ADP (final concentration 2 μmol l−1, catalog no. 0203001; mölab), Ala-Tyr-Pro-Gly-Lys-Phe-NH2 (final concentration 75 μM, catalog no. A3227; Sigma-Aldrich), U46619 (final concentration 5 μM, catalog no. 1932; Tocris Bioscience) and collagen (final concentration 1 μg ml−1, catalog no. 0203009). Platelet aggregation was recorded over 5 min to measure the area under the curve and displayed as a.u. min.
7
Neutrophil Isolation and Characterization
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7.5ml of blood were collected and distributes as follows:
Neutrophil counts were measured as part of the complete blood cell counts using a Sysmex XP-300 automated haematology analyser, (USA) following the manufacturer’s instruction. Eightcheck-3wp (USA) control samples were tested before running the patient samples.
Neutrophils were isolated as described in [27 (link)]. Briefly, following density gradient centrifugation on HistopaqueH-1077 (Sigma), the supernatant was discarded, and the pellet (containing erythrocytes and neutrophils) was resuspended in PBS; 3% dextran sulfate was added, and the tube was left to stand for 20min at room temperature to allow for erythrocyte sedimentation. The supernatant was collected, washed, and suspended in red cell lysis buffer and incubated at 4°C for 15min. The suspension was washed with PBS and the neutrophil sediment was resuspended in PBS.
Neutrophil counts were measured as part of the complete blood cell counts using a Sysmex XP-300 automated haematology analyser, (USA) following the manufacturer’s instruction. Eightcheck-3wp (USA) control samples were tested before running the patient samples.
Neutrophils were isolated as described in [27 (link)]. Briefly, following density gradient centrifugation on HistopaqueH-1077 (Sigma), the supernatant was discarded, and the pellet (containing erythrocytes and neutrophils) was resuspended in PBS; 3% dextran sulfate was added, and the tube was left to stand for 20min at room temperature to allow for erythrocyte sedimentation. The supernatant was collected, washed, and suspended in red cell lysis buffer and incubated at 4°C for 15min. The suspension was washed with PBS and the neutrophil sediment was resuspended in PBS.
8
Comprehensive Serum and Hematological Profiling
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Biochemical parameters of serum, including blood glucose (Glu), cholesterol (Ch), triglyceride (Tg), urea (Ur), creatinine (Cr), AST (aspartate aminotransferase or SGOT, serum glutamic-oxaloacetic transaminase), ALT (alanine aminotransferase or SGPT, serum glutamic pyruvic transaminase), ALP (alkaline phosphatase), calcium (Ca), and phosphorus (Ph) were measured using an auto-analyzer (BT 1500, Biotechnica Co., Italy).
Whole blood was evaluated for hematological parameters (complete blood count [CBC]) by an automated cell counter (XP-300, Sysmex Co., Japan). CBC included the number of white blood cells (WBCs), percentage of neutrophils (%Neu), red blood cells (RBCs), and their indexes, including mean cell volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), and red cell distribution width (RDW). Hemoglobin (Hb), hematocrit (Hct), and platelet counts (Plts) were also obtained.
Whole blood was evaluated for hematological parameters (complete blood count [CBC]) by an automated cell counter (XP-300, Sysmex Co., Japan). CBC included the number of white blood cells (WBCs), percentage of neutrophils (%Neu), red blood cells (RBCs), and their indexes, including mean cell volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), and red cell distribution width (RDW). Hemoglobin (Hb), hematocrit (Hct), and platelet counts (Plts) were also obtained.
9
Whole-Blood Leukocyte and Lymphocyte Analysis
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Leukocytes and lymphocytes were measured in EDTA whole-blood samples with a SYSMEX XP-300 differential haematology analyser.
10
Biomarker Discovery in Cancer Patients
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Venous blood samples from healthy volunteers or cancer patients were collected into blood collection tubes containing different anticoagulants and/or preservatives. For proteomics and RNA sequencing experiments, samples were collected from prostate cancer patients prior to local treatment. All other experiments were performed on healthy donor samples. All donors were non‐smokers. Healthy donors did not take any medication, nor did they suffer from any chronic or acute disease at the time of venipuncture. Cancer patients were not on anticoagulant therapy at the time of sample collection. For all donors, erythrocyte count, haemoglobin, mean cellular volume (MCV), mean cellular haemoglobin (MCH), mean cellular haemoglobin concentration (MCHC), platelet count and leukocyte count were documented by a haematology analyser (XP‐300, Sysmex, Kobe, Japan). Parameters were within the normal range for all included donors. Collection of biological samples was according to the Ethical Committee of Ghent University Hospital approval EC/2015/0260 and in accordance with the guidelines and regulations of the Helsinki Declaration. Participants had given written informed consent. Donor characteristics and clinical chemistry data are summarized in Table S1 . Clinical data from cancer patients are summarized in Table S2 .
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