Hemacytometer

Manufactured by Hausser Scientific

Sourced in United States, Panama

A hemacytometer is a device used to count and determine the concentration of cells, such as red blood cells or white blood cells, in a liquid sample. It consists of a thick glass or plastic slide with a counting chamber that is precisely calibrated to a known volume. The sample is placed in the counting chamber, and the cells are then counted under a microscope. The cell concentration can be calculated based on the number of cells counted and the volume of the sample.

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Lab products found in correlation

Trypan blue, by Thermo Fisher Scientific (4 mentions) Trypan blue dye, by Merck Group (2 mentions) Frosted glass slides, by Leica (2 mentions) Leishman solution, by Leica (2 mentions) Prism 6, by GraphPad (2 mentions) Spectramax plate reader, by Molecular Devices (1 mentions) Celltiter 96 aqueous one solution reagent, by Promega (1 mentions) Hi fbs, by Thermo Fisher Scientific (1 mentions) Vacutainer cpt mononuclear cell preparation tube, by BD (1 mentions) Penicillin streptomycin solution, by Thermo Fisher Scientific (1 mentions) Hepes, by Corning (1 mentions) Rpmi 1640 with l glutamine, by Corning (1 mentions) Celltiter glo luminescent viability assay, by Promega (1 mentions) Infinite m1000 pro plate reader, by Tecan (1 mentions) Phase contrast microscope, by Thermo Fisher Scientific (1 mentions) Phosphate buffered saline (pbs), by Corning (1 mentions) Endogro, by Merck Group (1 mentions) Mesenpro rs, by Thermo Fisher Scientific (1 mentions) Vegf165, by Thermo Fisher Scientific (1 mentions) Trypan blue, by Lonza (1 mentions)

Spelling variants of this product

Bright-Line Hemacytometer (17 citations) Neubauer hemacytometer (3 citations) Reichert Bright-Line Hemacytometer (3 citations) Gridded hemacytometer (2 citations)

30 protocols using hemacytometer

Cytotoxicity of Cyclocreatine in Myl and Myl-R Cells

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Myl and Myl-R cells maintained in normal growth medium (10% FBS) or regular media containing various concentrations of cyclocreatine (CCr), a competitive inhibitor of creatine known to have high affinity for the Na⁺/creatine symporter^{31 (link)}. Cyclocreatine was dissolved in regular cell growth medium at a stock concentration of 100 mM. In triplicate, CCr was added to 10 × 10³/100 μL Myl or Myl-R cells in increasing concentrations in a 96-well plate. Regular cell culture medium was used as the vehicle control. After 48-hr incubation at 37°C with 5% CO₂, cell viability was determined by MTS assay performed according to the manufacturer’s instructions (CellTiter 96® AQueous One Solution Reagent, Promega, Madison, WI). The absorbance was read at 490 nm using a SpectraMAX plate reader (Molecular Devices, Sunnyvale, CA). Trypan blue exclusion assay was performed using the Trypan blue reagent, and cells were counted using a hemacytometer (Hausser Scientific, Horsham, PA). Triplicate counts were obtained and used to determine cell proliferation. Data was analyzed for significant differences using a paired t-test.

Okumu D.O., Aponte-Collazo L.J., Dewar B.J., Cox N.J., East M.P., Tech K., McDonald I.M., Tikunov A.P., Holmuhamedov E., Macdonald J.M, & Graves L.M. (2019). Lyn regulates creatine uptake in an imatinib-resistant CML cell line. Biochimica et biophysica acta. General subjects, 1864(4), 129507.

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Blood Sample Processing and Preservation

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At each of the study sites, venous blood was collected from consented participants into BD Vacutainer CPT Mononuclear Cell Preparation Tubes (BD Biosciences) with sodium citrate. Tubes with separated blood samples were centrifuged for 10 minutes at 2000 rpm on site, and then stored on ice for ~ 8 hours during transportation to the University of Florida research lab in Gressier, Haiti. Both plasma and PBMCs were then separated via further centrifugation for 10 minutes at 2000 rpm. Plasma was decanted and stored at -80°C. Cells were enumerated using 0.4% trypan blue (Thermo Fisher Scientific) and a hemacytometer (Hausser Scientific) and stored at -80°C in RPMI 1640 with L-glutamine and 25mM HEPES (Corning Mediatech) supplemented with 10% heat-inactivated Hi-FBS (Gibco Life Technologies) and Penicillin/Streptomycin solution (Gibco Life Technologies) with 10% molecular grade DMSO. Samples were shipped on dry ice via air courier to the Emerging Pathogens Institute at the University of Florida, Gainesville, FL. Upon arrival to the Emerging Pathogens Institute, plasma was stored at -80°C and PBMCs were stored in liquid nitrogen until used in experiments.

Lehmann J.S., Campo J.J., Cicéron M., Raccurt C.P., Boncy J., Beau De Rochars V.E, & Cannella A.P. (2017). T cell subtypes and reciprocal inflammatory mediator expression differentiate P. falciparum memory recall responses in asymptomatic and symptomatic malaria patients in southeastern Haiti. PLoS ONE, 12(4), e0174718.

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Euthanasia and BALF Analysis

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Animals were euthanized 24 h after the last e-cigarette aerosol exposure by administering 2% of xylazine and 10% ketamine through i.p. injection. The collection of BALF was done as described previously and then centrifuged for 5 min. The supernatant was collected to measure the inflammatory mediators. The pellet was suspended in normal saline and hemacytometer (Hausser Scientific, Horsham, PA) was used to count the total inflammatory cells. A cytospin obtained from Cytotek (Netherlands) was used to perform the differential cell count. Cells were stained using Wright-Giemsa procedure. As well, the lungs were extracted and frozen immediately in liquid nitrogen [20] (link).

Alzoubi K.H., Khabour O.F., Al-Sawalha N.A., Karaoghlanian N., Shihadeh A, & Eissenberg T. (2022). Time course of changes in inflammatory and oxidative biomarkers in lung tissue of mice induced by exposure to electronic cigarette aerosol. Toxicology Reports, 9, 1484-1490.

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Trypan Blue Cell Viability Assay

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To quantify the percentage of dead cells, trypan blue cell impermeable dye was incubated with treated cells [26] . A 1∶1 mixture of cell suspension and trypan blue dye (Sigma-Aldrich, ON, Canada) was pipetted into a hemacytometer (Hausser Scientific, Horsham, PA) and the number of cells (dead cells stained blue and viable cells remained unstained) were manually quantified. The number of dead cells was expressed as a percentage of the total number of cells. Results were analyzed using GraphPad Prism 6.0 and reported as the mean ± SD of two independent experiments.

Larocque K., Ovadje P., Djurdjevic S., Mehdi M., Green J, & Pandey S. (2014). Novel Analogue of Colchicine Induces Selective Pro-Death Autophagy and Necrosis in Human Cancer Cells. PLoS ONE, 9(1), e87064.

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Cell Viability Assay Protocol

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Cells were resuspended and plated at a concentration of 2 × 10⁵ cells in 200 μl in 96-well tissue culture plates. Media with peptides was replaced every 48 h. To assess the number of viable cells, cells were resuspended in PBS and 10 µl of suspension was mixed in a 1:1 ratio with 0.4% Trypan Blue (Thermo Fisher) and counted using a hemacytometer (Hausser Scientific, Horsham, PA, USA). To assess viability using an ATP-based assay, cell viability was assessed using the CellTiter-Glo Luminescent Viability assay, according to the manufacturer’s instructions (Promega). Luminescence was recorded using the Infinite M1000Pro plate reader using integration time of 250 ms (Tecan).

Ramaswamy K., Forbes L., Minuesa G., Gindin T., Brown F., Kharas M.G., Krivtsov A.V., Armstrong S.A., Still E., de Stanchina E., Knoechel B., Koche R, & Kentsis A. (2018). Peptidomimetic blockade of MYB in acute myeloid leukemia. Nature Communications, 9, 110.

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Cell Viability Assessment by Trypan Blue

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Briefly, cells were harvested using trypsin and stained with 0.4% trypan blue dye (Sigma-Aldrich). Trypan blue-positive and -negative cells were counted using a hemacytometer (Hausser Scientific, Horsham, PA) under a phase-contrast microscope (Fisher Scientific, Pittsburgh, PA). The results of each assay were expressed in terms of the percentage of dead cells relative to the total number of cells. Individual experiments were performed in triplicate.

Chang Y.W., Su C.M., Su Y.H., Ho Y.S., Lai H.H., Chen H.A., Kuo M.L., Hung W.C., Chen Y.W., Wu C.H., Chen P.S, & Su J.L. (2014). Novel peptides suppress VEGFR-3 activity and antagonize VEGFR-3-mediated oncogenic effects. Oncotarget, 5(11), 3823-3835.

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Cell Growth Kinetics Measurement

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All cell lines were grown in HL5 medium until a density of 1–3 × 10⁴ cells/ mL was obtained. Cellular growth was determined by counting the cells every 24 h over five days using a hemacytometer (Hausser Scientific, Horshman, PA, USA). Experiments were performed in triplicate.

Yarbrough A., Maringer K., Saheb E.J., Jawed S, & Bush J. (2018). The Effect of Overexpressed DdRabS on Development, Cell Death, Vesicular Trafficking, and the Secretion of Lysosomal Glycosidase Enzymes. Biology, 7(2), 33.

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Intranasal Infection with Cryptococcus neoformans

Cited 1 time

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All experiments were conducted with a serotype D strain of Cryptococcus neoformans, 52D (ATCC 24067). This strain has been used extensively for studies of CN pathogenesis and virulence in mice. Aliquots of CN were stored at -80°C in 15% glycerol until needed. CN inocula were grown in Difco Sabouraud dextrose broth (Becton Dickinson, Franklin Lakes, NJ) at 30°C while shaking at 150 rpm for 48h and washed twice in phosphate-buffered saline (PBS, Cellgro, Corning, NY). Viable cells were determined using a hemacytometer (Hausser Scientific, Horsham, PA). Mice were anaesthetized with isoflurane (Halocarbon, River Edge, NJ) and then infected intranasally with 2 × 10⁵ CFU of CN in 20 µl of PBS as described previously.¹¹ (link) Inocula were confirmed by plating on Sabouraud dextrose agar plates (BBL, Sparks, MD).

Dufaud C., Rivera J., Rohatgi S, & Pirofski L.A. (2017). Naïve B cells reduce fungal dissemination in Cryptococcus neoformans infected Rag1−/− mice. Virulence, 9(1), 173-184.

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Cell Count with Trypan Blue

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Different gene-modified cells (5 × 10⁴ cells/well) were seeded in a 6-well plate. After serum starvation for 16 h, fresh 10% FBS DMEM was added and incubated for 7 days. The cell pellet was suspended in 1 mL of phosphate buffered saline (PBS) mixed 1:1 with 0.4% trypan blue (Gibco BRL, Grand Island, NY, USA). A hemacytometer (Hausser Scientific, Horsham, PA, USA) was used to count the cells.

Wang Y.H., Liu C.L., Chiu W.C., Twu Y.C, & Liao Y.J. (2019). HMGCS2 Mediates Ketone Production and Regulates the Proliferation and Metastasis of Hepatocellular Carcinoma. Cancers, 11(12), 1876.

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Carboxylated Particle Characterization

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PLGA, PLA, and PCL carboxylated 500 nm particles were obtained from Phosphorex, Inc. (Hopkinton, MA). The VTC particle concentration was obtained by manual counting on a hemacytometer (Hausser-Scientific). Non-fluorescent particles were used to limit any effect of a fluorescent dye on VTC-protein interaction. Particles were rendered fluorescent after the plasma/serum/VB incubation and prior to use in flow assays. Si and PS green fluorescent particles were purchased from Corpuscular (Cold Spring, NY) and Polysciences, Inc. (Warrington, PA), respectively. VTC size was measured by dynamic light scattering (DLS) using a Malvern Zetasizer instrument. Carboxylated stocks were dispersed in PBS++, with 1% bovine serum albumin (BSA) and then washed with 50 mM PBS prior to making the DLS measurement. Carboxylated biodegradable PLGA, PLA, and PCL particles were soaked in 50 mM MES at pH 7 (for PCL, pH ~5) for ≥ 20 hr prior to DLS measurement which corresponds to the time required for NeutrAvidin conjugation to these particles. VTC diameters ranged from ~400–700 nm as listed in Table 1. Table 2 lists the average sLe^a ligand site density of the various VTC materials.

Sobczynski D.J, & Eniola-Adefeso O. (2016). Effect of anticoagulants on the protein corona-induced reduced drug carrier adhesion efficiency in human blood flows. Acta biomaterialia, 48, 186-194.

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