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Neubauer hemocytometer

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The Neubauer hemocytometer is a laboratory instrument used for counting cells, particularly blood cells and other microscopic particles. It consists of a thick glass slide with a precisely etched grid pattern that creates a known volume, allowing for the calculation of cell concentration in a sample.

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Lab products found in correlation

Accutase, by Thermo Fisher Scientific (1 mentions) Trypan blue, by Merck Group (1 mentions) Dulbecco s phosphate buffered saline dpbs, by Merck Group (1 mentions) Image pro plus 6, by Media Cybernetics (1 mentions) Rpmi 1640, by Thermo Fisher Scientific (1 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions) Hank s balanced salt solution, by Thermo Fisher Scientific (1 mentions) Ang 2, by R&D Systems (1 mentions) Coomassie protein assay kit, by Thermo Fisher Scientific (1 mentions) Mip 2, by R&D Systems (1 mentions) Propidium iodide, by Thermo Fisher Scientific (1 mentions)

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Hemocytometer (123 citations) Neubauer hemocytometer chamber (5 citations)

6 protocols using neubauer hemocytometer

1

Cell Detachment and Viability Assay

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At the end of the experiments, adherent cells were washed with 0.5 mL/well of ice-cold Dulbecco’s phosphate buffered saline (DPBS; Sigma-Aldrich), incubated 15 min at RT in 300 μL of an enzymatic cell detachment solution (Accutase®; Thermo Fisher Scientific), and detached by gentle pipetting using a 1-mL manual single-channel pipette (Mettler Toledo, Columbus, OH). The cell suspensions were transferred into 2-mL untreated polystyrene culture tubes (Axygen Scientific, Union City, CA), and cell mortality was analyzed by dye-exclusion hemocytometry under phase contrast microscopy using trypan blue (0.04% [w/v] final concentration; Sigma-Aldrich) and an improved Neubauer hemocytometer (Hausser Scientific, Horsham, PA).
Ferko M.A, & Catelas I. (2018). Effects of metal ions on caspase-1 activation and interleukin-1β release in murine bone marrow-derived macrophages. PLoS ONE, 13(8), e0199936.
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2

Quantitative Cell Counting and Sizing

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In each experiment, five samples from each flask and from each treatment were counted at each sampling period. C. sorokiniana cells were counted under a light microscope, using a Neubauer hemocytometer (bright line counting chamber, Hausser Scientific, Horsham, PA) connected to an image analyzer (Image ProPlus 6.3, Media Cybernetics, Silver Spring, MD). A. brasilense Cd, B. pumillus ES4, and E. coli DH5α were counted after serial dilution by the plate count method on nutrient agar medium (M7519, Sigma-Aldrich, St. Louis, MO). Cell volume of C. sorokiniana was measured by the same image analyzer. Five samples per treatment per each sampling time were analyzed and each was analyzed by five microscopic fields (n = 50 individual analyses). The volume of spherical cells was calculated.
Amavizca E., Bashan Y., Ryu C.M., Farag M.A., Bebout B.M, & de-Bashan L.E. (2017). Enhanced performance of the microalga Chlorella sorokiniana remotely induced by the plant growth-promoting bacteria Azospirillum brasilense and Bacillus pumilus. Scientific Reports, 7, 41310.
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3

Cell Culture Protocols for Adherent and Suspension Cells

Cited 2 times
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Cell lines were obtained from the German Collection of Microorganisms and Cell Cultures (Brunswick, Germany). All cells but retinal pigment epithelial (RPE) were cultured in RPMI-1640 medium (Invitrogen, Grand Island, NY) supplemented with penicillin/streptomycin and 10% fetal bovine serum. RPE cells were cultured in DMEM (Invitrogen, Grand Island, NY) medium supplemented with penicillin/streptomycin and 10% fetal bovine serum. Suspension and adherent cells were maintained at a density of 0.5–1 million cells/ml or 50–90% confluence, respectively, at 37 ºC in a humidified atmosphere with 5% CO2. Jurkat cells engineered to express GFP were generated by retroviral transduction as described.23 (link)–24 (link) Cell counts were measured using the Neubauer hemocytometer (Hausser Scientific, Horsham, PA), according to manufacturer’s instructions.
Dhabaria A., Cifani P., Reed C., Steen H, & Kentsis A. (2015). A high-efficiency cellular extraction system for biological proteomics. Journal of proteome research, 14(8), 3403-3408.
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4

Cell Counting Using Hemocytometer

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The cell count was performed using a Neubauer hemocytometer (Hausser Scientific Company, Horsham, PA). Thirty microliters of the suspended pellet was taken and spread on the counter slide. The number of cells in four large squares (1 mm2) was counted to determine the total cell count. The concentration of the cells from the uterine horn and oviduct were determined.
, & Aldarmahi A. (2016). Establishment and characterization of female reproductive tract epithelial cell culture. Journal of Microscopy and Ultrastructure, 5(2), 105-110.
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5

Bronchoalveolar Lavage Cell Analysis

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Immediately after collection of BAL fluids, erythrocytes were lysed using 0.2% saline and the remaining cells were resuspended in Hanks' Balanced Salt Solution (Invitrogen, Grand Island, NY). Total cell count of each BAL sample was determined using a Neubauer hemocytometer (Hausser Scientific, Horsham, PA). Differential cell counts were performed (200 cells for each experimental condition) on cytospin slides stained with Protocol HEMA3 solution (Fisher scientific, Fair Lawn, NJ). MPO levels were determined using Suzuki’s protocol [17 (link)]. Total protein concentration in BAL fluid was measured using Coomassie protein assay kit (Thermo Scientific, Rockford, IL). Ang-2, KC and MIP-2 concentration in plasma, tissue homogenates and BAL fluid was measured using specific ELISAs (Ang-2, R&D Systems, Minneapolis, MN; KC and MIP-2, Millipore Inc., Billerica, MA) according to the manufacturer’s instructions.
Lee J.Y., Linge H.M., Ochani K., Lin K, & Miller E.J. (2016). N-Ethylmaleimide Sensitive Factor (NSF) Inhibition Prevents Vascular Instability following Gram-Positive Pulmonary Challenge. PLoS ONE, 11(6), e0157837.
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6

Dual Fluorescent Viability Assay

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Cells were tested for viability using acridine orange (AO, Life Technologies, Carlsbad, CA) and propidium iodide (PI, Life Technologies, Carlsbad, CA). A working solution of AO/PI was made by mixing 1.0 mL of 1mM AO stock in sterile water, 2.0mL of 0.5 mg/mL PI in DPBS, and 47mL of DPBS. The final working solution with concentrations of 20μM AO and 20μg/mL PI was combined with cells in a 1:15–1:20 dilution. At least 100 cells were counted in a Neubauer hemocytometer (Hausser Scientific, Horsham, PA) under fluorescence. Under these conditions, viable live cells fluoresced green due to AO binding to nucleic acids via intercalation. Conversely, PI caused dead cells to fluoresce red, as it is membrane impermeant in healthy cells. Sample viability percentage was calculated by dividing the number of live cells by the number of total cells (live+dead) and multiplying by 100.
Pollock K., Sumstad D., Kadidlo D., McKenna D.H, & Hubel A. (2014). Clinical mesenchymal stem cell products experience functional changes in response to freezing. Cytotherapy, 17(1), 38-45.
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