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Act diff analyzer

Manufactured by Beckman Coulter
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The AcT diff Analyzer is a compact, automated hematology analyzer designed for rapid and accurate analysis of complete blood count (CBC) and white blood cell differential (DIFF). It provides precise measurements of red blood cells, white blood cells, and platelet counts, as well as the distribution of different types of white blood cells.

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Lab products found in correlation

Lsrfortessa, by BD (4 mentions) Facscanto 2, by BD (2 mentions) Facsymphony, by BD (2 mentions) Ficoll paque premiem, by GE Healthcare (2 mentions) Collagenase d, by Roche (2 mentions) Alt activity assay kit, by Merck Group (1 mentions) Ast activity assay kit, by Merck Group (1 mentions) 21g needle, by BD (1 mentions) 7 aad, by BioLegend (1 mentions) Annexin 5 af647, by BioLegend (1 mentions) Facscanto, by BD (1 mentions) Zombie fixable viability kit, by BioLegend (1 mentions) Live dead fixable dead cell stain kit, by Thermo Fisher Scientific (1 mentions) Ack lysis buffer, by Thermo Fisher Scientific (1 mentions) Abx pentra 400, by Horiba (1 mentions) Lymphoprep, by STEMCELL (1 mentions) Ficoll paque, by GE Healthcare (1 mentions) Rx daytona, by Randox (1 mentions)

Spelling variants of this product

AcT/Diff (20 citations) Coulter AcT diff Analyzer (19 citations) AcT Diff Hematology Analyzer (13 citations) AcT 5 diff AL Hematology Analyzer (8 citations) ACT 5 Diff. hematology analyzer (8 citations)

14 protocols using act diff analyzer

1

Whole Blood Analysis and Liver Function in Mice

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For whole blood analysis, mice were bled from the facial vein using a lancet. Blood was collected in EDTA tubes to prevent clotting (BD Biosciences), briefly vortexed and whole blood analysis was carried out using an ACT diff analyzer (Beckman Coulter) immediately after blood collection. For each blood sample, two analytical readings were taken and averaged. ALT Activity Assay kit (MAK052, Sigma-Aldrich) and AST Activity Assay kit (MAK055, Sigma-Aldrich) were used to perform liver function tests.
Rathore A.P., Mantri C.K., Tan M.W., Shirazi R., Nishida A., Aman S.A., Morrison J, & St. John A.L. (2021). Immunological and Pathological Landscape of Dengue Serotypes 1-4 Infections in Immune-Competent Mice. Frontiers in Immunology, 12, 681950.
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2

Blood Biomarkers Evaluation Protocol

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Fasting blood samples (following 48 h rest from training) were collected into an ethylenediaminetetraacetic acid (EDTA) tube from an antecubital vein using a 21G needle (BD Diagnostics, Dublin, Ireland) at pre- and post-intervention testing, subsequent to a 12-h overnight fast. Blood samples were put through a haematology analyser (Beckman Coulter AcT diff Analyzer, Beckman Coulter, Brea, CA, USA) 20–30 min subsequent to collection. The haematology analyser measured participant’s white blood cells, lymphocytes, monocytes, granulocytes, red blood cells, haematocrit, mean corpuscular volume (MCV), mean corpuscular width (MCW), red blood cell distribution width (RDW) and platelet content. Serum ferritin was measured pre and post intervention via Enzyme Linked Immunosorbent Assay (ELISA) (GmbH, Aachen, Germany). Intra-assay coefficient of variation was 3.9–9.9 ng/mL.
McSwiney F.T, & Doyle L. (2019). Low-Carbohydrate Ketogenic Diets in Male Endurance Athletes Demonstrate Different Micronutrient Contents and Changes in Corpuscular Haemoglobin over 12 Weeks. Sports, 7(9), 201.
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3

Multiparametric Flow Cytometry Analysis

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All fluorescently labeled antibodies were purchased from BD Biosciences, BioLegend and Invitrogen (Thermo Fisher Scientific) (S1 Table). Lysis of erythrocytes in whole blood was done with NH4Cl. Spleens were mechanically disrupted and filtered through a 70 μm cell strainer before separation of mononuclear cells on Ficoll-Paque gradients. Cell suspensions were stained with antibodies for 30 min on ice, washed, and analyzed on FACSCanto or LSR Fortessa cytometers (BD Biosciences). Analysis of flow cytometric data was performed with FlowJo (Tree Star). The absolute numbers of leukocytes in each category were computed from the white blood cell count measured with a Beckman Coulter AcT diff Analyzer. Early apoptotic cells were assessed by incubating cells with AF647 annexin V (BioLegend) and 7AAD (BioLegend) diluted in annexin binding buffer for 15 min at room temperature in the dark, 400 μl binding buffer was added and the staining was directly analyzed on FACSCanto.
Murer A., McHugh D., Caduff N., Kalchschmidt J., Barros M., Zbinden A., Capaul R., Niedobitek G., Allday M., Chijioke O, & Münz C. (2018). EBV persistence without its EBNA3A and 3C oncogenes in vivo. PLoS Pathogens, 14(4), e1007039.
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4

Phenotyping Immune Cells in huNSG-A2 Mice

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Whole blood and splenocyte cell suspensions of huNSG-A2 mice were subjected to erythrocyte lysis by ACK lysis buffer (Life Technologies) prior to staining. Fluorescently-labeled antibodies were purchased from BioLegend unless otherwise stated: CD45 (Pacific Blue/HI30), CD3 (PE/UCHT1 or BV785/OKT3), CD8 (PerCP/SK1), CD4 (APC-Cy7/RPA-T4 or BV605/OKT4), CD19 (PE-Cy7/HIB19 or AF700/HIB19), HLA-DR (FITC/L243), CD62L (APC/DREG-56, BD Bioscience) and CD45RA (BV510/HI100). Cell viability was assessed using fixable NIR and Aqua viability dyes (Zombie Fixable Viability Kit, BioLegend or LIVE/DEAD Fixable Dead Cell Stain Kit, ThermoFisherScientific). Acquisition was performed on BD FACSCanto II or BD LSR Fortessa flow cytometers and data was analyzed using FlowJo 9.9 software. Absolute leukocyte numbers were calculated from white blood cell counts measured with a hemocytometer (Beckman Coulter AcT Diff Analyzer).
Caduff N., McHugh D., Murer A., Rämer P., Raykova A., Landtwing V., Rieble L., Keller C.W., Prummer M., Hoffmann L., Lam J.K., Chiang A.K., Raulf F., Azzi T., Berger C., Rubic-Schneider T., Traggiai E., Lünemann J.D., Kammüller M, & Münz C. (2020). Immunosuppressive FK506 treatment leads to more frequent EBV-associated lymphoproliferative disease in humanized mice. PLoS Pathogens, 16(4), e1008477.
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5

Hematology Analysis of Blood Samples

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Blood samples were analyzed on a Coulter Ac•T diff Analyzer using the Veterinary Application Software (Beckman-Coulter, Fullerton, CA) and reticulocytes were counted at the Pathology Department of the Royal Brisbane and Women's Hospital (Brisbane, Australia) using a Sysmex XE-5000 automated hematology analyzer (Roche Diagnostics, Castle Hill, NSW, Australia).
Fuqua B.K., Lu Y., Darshan D., Frazer D.M., Wilkins S.J., Wolkow N., Bell A.G., Hsu J., Yu C.C., Chen H., Dunaief J.L., Anderson G.J, & Vulpe C.D. (2014). The Multicopper Ferroxidase Hephaestin Enhances Intestinal Iron Absorption in Mice. PLoS ONE, 9(6), e98792.
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6

Multiparametric Analysis of Hematopoietic Cells

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Peripheral blood from mice was collected either by tail vein bleeding or terminal heart puncture at the day of sacrifice. An aliquot of whole blood was used to determine the whole blood cell count with a Beckman Coulter AcT diff Analyzer for the calculation of total cell counts. Red blood cells were lysed with ACK-lysis buffer for 5 min and subsequently stained with fluorochrome-conjugated antibodies for phenotypic analysis by flow cytometry. Cells were stained with surface markers for 20 min at 4 °C. Human reconstitution levels were determined with the expression of human CD45, CD3, CD4, CD8, CD19, and NKp46. Labeled cells were acquired on a BD FACSCantoII, BD LSRFortessa, or BD FACSymphony. Data analysis was performed using FlowJo software (FlowJo LLC).
Engelmann C., Schuhmachers P., Zdimerova H., Virdi S., Hauri-Hohl M., Pachlopnik Schmid J., Grundhoff A., Marsh R.A., Wong W.W, & Münz C. (2022). Epstein Barr virus-mediated transformation of B cells from XIAP-deficient patients leads to increased expression of the tumor suppressor CADM1. Cell Death & Disease, 13(10), 892.
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7

Tissue Dissociation and Cell Isolation

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Spleen tissue was manually dissociated, filtered through 70-μm cell strainer, and subjected to Ficoll gradient centrifugation (Ficoll Paque Premiem, GE Healthcare). Liver tissue was digested for 30min at 37°C in buffer containing DNase (0.4mg/ml) and Collagenase D (20μg/ml) (Roche), followed by addition of 0.5M EDTA, filtering and separation of mononuclear cells on Percoll gradients. Whole blood was collected by tail-vein and terminal cardiac puncture in tubes containing heparin or EDTA. Prior to erythrocyte lysis by ACK lysis buffer, the white blood cell count in whole blood was determined with a hematology analyzer (Beckman Coulter AcT Diff Analyzer). Human PBMCs were isolated from Lithium-Heparin vacutainers by Ficoll gradient centrifugation within 2 hours of the blood draw and cryopreserved until use, as described by Forconi et al. (2018) (link).
Tenover F.C., Jones R.N., Swenson J.M., Zimmer B., McAllister S, & Jorgensen J.H. (1999). Methods for Improved Detection of Oxacillin Resistance in Coagulase-Negative Staphylococci: Results of a Multicenter Study. Journal of Clinical Microbiology, 37(12), 4051-4058.
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8

Isolation of CD8+ and CD19+ Cells

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PBMCs were obtained from humanized mice by lysing blood for five minutes with 1x ACK lysis buffer, followed by washing. Spleens were manually dissociated, and cells filtered through a 70 μm strainer before separation of white blood cells on Ficoll-Paque gradients. Cells were counted using a Beckman Coulter AcT diff Analyzer. CD8+ and CD19+ cells were isolated via positive selection using Miltenyi microbeads and following manufacturer’s recommendations.
Chatterjee B., Deng Y., Holler A., Nunez N., Azzi T., Vanoaica L.D., Müller A., Zdimerova H., Antsiferova O., Zbinden A., Capaul R., Dreyer J.H., Nadal D., Becher B., Robinson M.D., Stauss H, & Münz C. (2019). CD8+ T cells retain protective functions despite sustained inhibitory receptor expression during Epstein-Barr virus infection in vivo. PLoS Pathogens, 15(5), e1007748.
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9

Comprehensive Blood Analysis Protocol

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Hematological analysis was performed using a Beckman Coulter Ac-T diff analyzer (Beckman Coulter, High Wycombe, United Kingdom). Total protein, blood urea nitrogen, cholesterol, glucose, triglycerides, creatinine, and creatine kinase concentrations were determined in serum samples using an ABX Pentra 400 clinical chemistry analyzer (Horiba, Northampton, United Kingdom) calibrated according to the manufacturer’s instructions, with every fifth sample run in duplicate to determine analyzer accuracy.
McCormack U.M., Curião T., Wilkinson T., Metzler-Zebeli B.U., Reyer H., Ryan T., Calderon-Diaz J.A., Crispie F., Cotter P.D., Creevey C.J., Gardiner G.E, & Lawlor P.G. (2018). Fecal Microbiota Transplantation in Gestating Sows and Neonatal Offspring Alters Lifetime Intestinal Microbiota and Growth in Offspring. mSystems, 3(3), e00134-17.
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10

PBMC Isolation and Characterization

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Peripheral blood mononuclear cells were isolated by gradient centrifugation as described38 (link). In brief, blood from healthy donors or patients with rheumatoid arthritis was diluted 1:1 in phosphate-buffered saline (PBS) and overlaid on Lymphoprep (Stem cell Technologies, Tullamarine, Victoria, Australia). Following density gradient separation PBMCs were collected and washed twice in sterile PBS and total and differential count performed using the AcT diff Analyzer (Beckman-Coulter). Cells were then immediately used for flow cytometry or treated with relevant recombinant cytokines in vitro for 0–18 h. All PBMCs were prepared and cultured under strict LPS-minimised conditions by using LPS-free culture media and reagents, z-filtration of buffers (<0.05 endotoxin units/ml), and by heat and/or chemical treatment of glassware and fume hoods (<0.2 units/item, Techni Tool, PA, USA).
Kane B.A., An H., Rajasekariah P., McNeil H.P., Bryant K, & Tedla N. (2019). Differential expression and regulation of the non-integrin 37/67-kDa laminin receptor on peripheral blood leukocytes of healthy individuals and patients with rheumatoid arthritis. Scientific Reports, 9, 1149.
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