Neubauer hemocytometer

Manufactured by Merck Group

Sourced in United States, Germany

The Neubauer hemocytometer is a laboratory instrument used for the counting and enumeration of cells, particularly blood cells. It consists of a microscope slide with a defined grid pattern etched onto its surface, allowing for precise cell counting and concentration measurements.

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Lab products found in correlation

Trypan blue, by Merck Group (4 mentions) Hepes and l amino acids, by Thermo Fisher Scientific (2 mentions) Fetal calf serum (fcs), by Thermo Fisher Scientific (2 mentions) Axiovert 25 microscope, by Zeiss (1 mentions) Trypsin 0.2 edta, by Thermo Fisher Scientific (1 mentions) Edta tube, by BD (1 mentions) Mouse fc block, by BD (1 mentions) Turk s solution, by Merck Group (1 mentions) Trypsin edta, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Kaluza 1, by Beckman Coulter (1 mentions) Gallios analyzer, by Beckman Coulter (1 mentions) Cytofix cytoperm buffer, by BD (1 mentions) Anti il 1β, by BD (1 mentions) Mem 41500, by Thermo Fisher Scientific (1 mentions) Observer d1, by Zeiss (1 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions) Dnase 1, by Merck Group (1 mentions) Collagenase type 4, by Merck Group (1 mentions) Rpmi 1640, by BD (1 mentions)

Spelling variants of this product

Hemocytometer (60 citations) Improved Neubauer hemocytometer (6 citations) BLAUBRAND® Neubauer Hemocytometer (2 citations)

22 protocols using neubauer hemocytometer

Fibroblast Expansion Protocol

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After the primary pre-culture was stabilized, 3×10^{5 (link)} fibroblasts were expanded in 150 cm² flasks at a density of 2,000
cells/cm². Standard cultures were maintained in a CO₂ incubator
(Heraeus BB 6060, Kendro, CORNING, Corning, NY, USA) at 37°C in a humidified atmosphere
with 5% CO₂. After cell isolation and during passages, cells were cultured in
MSCGM medium (LONZA). The medium was changed every 3 days. As 80% confluence was reached,
cells were detached using 0.05% trypsin 0.2% EDTA (Gibco, Thermofisher, Waltham, MA, USA),
and 3×10⁵ cells were passaged to a new flask. Cell suspensions were counted
using a Neubauer hemocytometer (Merck, Darmstadt, Germany). Culture quality control,
microbiology testing, and morphology examination was done with phase-contrast microscopy
using an Axiovert 25 microscope (Zeiss, Göttingen, Germany). The cell expansion rate (k)
was calculated using the cell doubling time formula
x_f=x₀e^(t*^k), where
x₀ is the initial number of seeded keratinocytes,
x_f is the final number of the harvested population, and
t is the time that the hFDFs were in culture.

Esteban-Vives R., Ziembicki J., Sun Choi M., Thompson R.L., Schmelzer E, & Gerlach J.C. (2019). Isolation and Characterization of a Human Fetal Mesenchymal Stem Cell Population: Exploring the Potential for Cell Banking in Wound Healing Therapies. Cell Transplantation, 28(11), 1404-1419.

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Murine Immune Cell Profiling by Flow Cytometry

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Blood was collected in EDTA tubes (BD Pharmingen, San Diego, CA, USA) and leukocytes counted in a Neubauer hemocytometer using Turk’s solution (Merck KGaA, Darmstadt, Germany). Lymphocytes and granulocytes were distinguished by morphology of nuclei. After lysis of red blood cells, FcγII and FcγIII receptors were blocked with anti-CD16/CD32 antibodies at 5 μg/ml (Mouse BD Fc Block™, BD Pharmingen) then specific primary antibodies were added: fluorescein isothiocyanate (FITC)-conjugated rat anti-mouse CD45R/B220 monoclonal antibody; phycoerythrin (PE)-conjugated hamster anti-mouse CD3e monoclonal antibody; PE-conjugated rat anti-mouse CD335 (NKp46) monoclonal antibody; FITC-conjugated rat anti-mouse CD49b (clone DX5) monoclonal antibody; FITC-conjugated rat anti-mouse CD8a monoclonal antibody; PE-conjugated rat anti-mouse CD4 monoclonal antibody; PE-conjugated rat anti-mouse CD44 monoclonal antibody. All antibodies were from BD Pharmingen. The Partec CyFlow® space cytofluorimeter was used and analysis was performed with Windows™ FloMax® software. Through backgating from the CD3+ and B220+ clusters, we selected the lymphocyte gate in the FSC-SSC dot plot. At least 10,000 events were acquired in the gate. The analysis template was kept constant in the various experiments. The same reagents and procedures were used for splenocyte immunophenotyping.

Croci S., Nanni P., Palladini A., Nicoletti G., Grosso V., Benegiamo G., Landuzzi L., Lamolinara A., Ianzano M.L., Ranieri D., Dall’Ora M., Iezzi M., De Giovanni C, & Lollini P.L. (2015). Interleukin-15 is required for immunosurveillance and immunoprevention of HER2/neu-driven mammary carcinogenesis. Breast Cancer Research : BCR, 17(1), 70.

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Evaluating Cytokine Production in F3II Cells

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Monolayer F3II cell cultures were treated with trypsin-EDTA (Gibco BRL) for 5 minutes at 37°C.After two washes with MEM 41500 (Gibco BRL) containing 10% FBS (Gibco BRL), the cells were counted using a Neubauer hemocytometer (Merck). A total of 2 × 10⁵ cells in 1 mL of PBS containing 1% bovine serum albumin were added to each well of a 96-well round (U) bottomplate. MHC I and II expression levels were determined after incubating the cells for 20 min on ice with anti-H-2K^dFITC and anti-IA/IE PE antibodies (BD Biosciences, Franklin Lakes, USA). Cytokine production was detected using intracellular staining. Cells were fixed and permeabilized with BD Cytofix/Cytoperm Buffers (BD Biosciences) and incubated with anti-interleukin (IL)-10, anti-IL-1β, anti-transforming growth factor (TGF)-β, anti-IL-6, or anti-tumor necrosis factor (TNF)-α antibodies. All antibodies were purchased from BD Biosciences. For each activation marker, an autofluorescence threshold was established by staining the control cells with the corresponding isotype controls. Staining was detected using a Gallios analyzer flow cytometer (Beckman Coulter, Brea, USA) and analyzed using the Kaluza 1.2 software (Beckman Coulter) for all immunofluorescent flow cytometry investigations.

Fuentes D., Cabezas-Cruz A., Mesa C., Carmenate T., Martínez D., Valdés-Zayas A., Montero E, & Pérez R. (2022). Murine Mammary Carcinoma Induces Chronic Systemic Inflammation and Immunosuppression in BALB/c Mice. Journal of Breast Cancer, 25(3), 218-232.

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Smartphone-based Semen Analysis: Cryopreservation and Counting

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Cryopreserved semen samples were obtained from California Cryobank and New England Cryobank and were stored in liquid nitrogen. The samples were thawed before use by placing them in an incubator at 37°C for 14 min. The samples were serially diluted with phosphate-buffered saline at multiple ratios. The samples were loaded into the microchips and were tested using the smartphone-based semen analyzer. In parallel, 10 μl of each sample was loaded into a Neubauer hemocytometer (Sigma, Z359629) and was counted under a phase-contrast optical microscope (Zeiss, Observer D1). At least four different FOVs were analyzed for each sample, and three repeated measurements were made using the smartphone application.

Kanakasabapathy M.K., Sadasivam M., Singh A., Preston C., Thirumalaraju P., Venkataraman M., Bormann C.L., Draz M.S., Petrozza J.C, & Shafiee H. (2017). An automated smartphone-based diagnostic assay for point-of-care semen analysis. Science translational medicine, 9(382), eaai7863.

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Isolation of Chicken Lymphocytes

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Five days before infection, each time point after infection, 5 d before immunization, and each time point after immunization, heparinized blood samples from individual chickens were collected to isolate peripheral blood lymphocytes (PBL) as previously described (Dai et al., 2020 ). Single cell suspensions of lung were obtained in RPMI 1640 medium (Gibco, Carlsbad, CA) according to the manufacturer's instructions of tissue mononuclear cell kit (Haoyang, Tianjin, China). Briefly, Dice lung lobes into slurry in petri dish using curved scissors while immersed in digestion media containing 0.002% DNase I (Sigma-Aldrich, St. Louis, MO) and 0.1% Collagenase Type 4 (Sigma-Aldrich) for 20 min at 37°C. Filter digested tissue through 190 μM metal mesh and then isolated lymphocytes with the tissue separation medium in the kit, as described before (Dai et al., 2020 ). Cell viability and counting was performed using Trypan Blue and a Neubauer hemocytometer (Sigma-Aldrich). And, the chicken PBL and tissue single cell suspensions were frozen in liquid nitrogen for later study.

Dai M., Li S., Keyi S.h.i., Sun H., Zhao L., Deshui Y.u., Liao J., Xu C, & Liao M. (2020). Comparative analysis of key immune protection factors in H9N2 avian influenza viruses infected and immunized specific pathogen–free chicken. Poultry Science, 100(1), 39-46.

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Establishing Ehrlich Ascites Carcinoma Model

Cited 1 time

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The Ehrlich ascites carcinoma (EAC) cell line was acquired from the Pharmacology and Experimental Oncology Unit of the National Cancer Institute (NCI), Cairo University, Egypt. Both solid and ascetic EAC cell growth was possible. The initial tumor for EAC cells was a spontaneous mouse breast cancer, and these cells are of mammary origin. Within ten days, Ehrlich tumor cells appeared in the ascetic fluid and were collected via IP puncture using a sterile syringe and dilution. The cells were counted using a Neubauer Hemocytometer (Sigma-Aldrich, St. Louis, MO, USA) [70 (link)]. The cells were determined to represent more than 99% of viability using the trypan blue dye exclusion technique [71 (link)]. Then, an SEC model in female Swiss albino mice was established. Briefly, 0.2 mL of live EAC cells (5 × 10⁶/mL) were injected subcutaneously in the left thigh of the lower limb and left for 12 days. A 14-day treatment cycle with several medications was started on the 12th day after injecting the EAC cells [72 (link),73 (link)].

Alamoudi J.A., El-Masry T.A., Nasr M., Ibrahim I.T., Ibrahim H.A., Saad H.M., El-Nagar M.M., Alshawwa S.Z., Alrashidi A, & El Zahaby E.I. (2024). Fabrication of Nanocrystals for Enhanced Distribution of a Fatty Acid Synthase Inhibitor (Orlistat) as a Promising Method to Relieve Solid Ehrlich Carcinoma-Induced Hepatic Damage in Mice. Pharmaceuticals, 17(1), 96.

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Spleen Isolation and Cell Preparation

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Example 9

Spleens were aseptically isolated and maintained in RPMI 1640 (Life Technologies, Darmstadt, Germany) at 4° C. until further processing. Spleen was passed through a 100 μm nylon cell strainer (BD Biosciences, Heidelberg, Germany) into 10 ml RPMI 1640 and cells were pelleted at 300×g for 5 min. For red blood cell lysis pellet was resuspended in 1 ml ACK Lysing solution (Life Technologies), incubated 10 min at room t° and centrifuged at 300×g for 5 min. Cells were resuspended in DPBS (Life Technologies) and cell concentration determined using Neubauer hemocytometer and Trypan blue (Sigma-Aldrich) staining for dead cell exclusion.

US10344067B2. RNA viruses expressing IL-12 for immunovirotherapy (2019-07-09). DEUTSCHES KREBSFORSCHUNGSZENTRUM [DE], RUPRECHT-KARLS UNIVERSITAET [DE]. Inventors: Guy Ungerechts [DE], Christine Engeland [DE], Ruta Veinalde [DE].

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Assessing Cell Lysis in Biotinylation

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Cell lysis during biotinylation was assessed by counting live cells before and after the biotinylation procedure. Cell numbers were assessed using trypan blue (Gibco, Invitrogen, Lofer, Austria) and a Neubauer hemocytometer (Sigma-Aldrich, Vienna, Austria).
Only samples with cell lysis below 10% after biotinylation were used for further analyses.

Ramires M.D., Hummel K., Hatfaludi T., Riedl P., Hess M, & Bilic I. (2022). Comparative Surfaceome Analysis of Clonal Histomonas meleagridis Strains with Different Pathogenicity Reveals Strain-Dependent Profiles. Microorganisms, 10(10), 1884.

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Quantifying Symbionts in Marine Invertebrates

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The density of the endosymbionts from the genus Philozoon sp. (LaJeunesse et al., 2021 (link)) was assessed according to Zamoum and Furla (2012) (link). Briefly, two tentacles were cut from each animal before and after H₂O₂ treatment in each condition (0, 200, or 500 μM H₂O₂) and then put in 2 M solution of NaOH and incubated at 37°C for 1 h to dissolve all animal tissue. To determine symbiont density, three replicate samples were counted using a modified Neubauer hemocytometer (Sigma-Aldrich). The remaining extract was used to determine the protein content from which we normalized symbiont density.

Cotinat P., Fricano C., Toullec G., Röttinger E., Barnay-Verdier S, & Furla P. (2022). Intrinsically High Capacity of Animal Cells From a Symbiotic Cnidarian to Deal With Pro-Oxidative Conditions. Frontiers in Physiology, 13, 819111.

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Isolation of Lymphocytes from Spleen and Liver

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For isolation of lymphocytes, spleen and liver initially collected in PBS + 2% FCS were transferred into petri dishes. The splenic capsule was removed and tissue teased apart by using two sterile blunt-end forceps. Liver tissue was dissected by squeezing the tissue with the end of a plunger from a 20 mL syringe. Cell suspensions from both organs were filtered through 40 µm cell strainers (BD Falcon™, BD Biosciences, San Jose, CA, USA). Spleen and liver cell suspensions were centrifuged at 350 × g for 10 min at room temperature and the supernatant was discarded. The cell pellet was resuspended with cold PBS + 2% FCS and layered on a double volume of Histopaque^®-1077 (Sigma-Aldrich). After centrifugation at 850 × g for 20 min at room temperature, the interphase was collected. Following washing with PBS and centrifugation at 650 × g for 10 min at 4 °C, cells were resuspended in PBS and stored on ice until further processing. Cell viability and counting was performed using Trypan Blue and a Neubauer hemocytometer (Sigma-Aldrich).

Lagler J., Mitra T., Schmidt S., Pierron A., Vatzia E., Stadler M., Hammer S.E., Mair K.H., Grafl B., Wernsdorf P., Rauw F., Lambrecht B., Liebhart D, & Gerner W. (2019). Cytokine production and phenotype of Histomonas meleagridis-specific T cells in the chicken. Veterinary Research, 50, 107.

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