Guava easycyte ht flow cytometer

Manufactured by Merck Group

Sourced in United States, Germany, France

The Guava easyCyte HT flow cytometer is a compact and automated benchtop instrument used for cellular analysis. It employs flow cytometry technology to detect and quantify various cellular parameters, such as size, granularity, and fluorescence. The Guava easyCyte HT is designed for efficient and reliable data acquisition and analysis.

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Lab products found in correlation

Guava incyte software, by Merck Group (3 mentions) Icam 1 cd54, by BD (2 mentions) Fluorochrome conjugated mouse igg isotype control, by BD (2 mentions) Endoglin cd105, by BD (2 mentions) Pecam1 cd31, by BD (2 mentions) Guava cell cycle reagent, by Merck Group (2 mentions) Click it plus opp protein synthesis assay kit, by Thermo Fisher Scientific (2 mentions) Opp reagent, by Jena Biosciences (2 mentions) Annexin 5 apc pi staining kit, by BioLegend (2 mentions) Pft α, by Merck Group (2 mentions) Annexin 5 apoptosis detection kit apc, by Thermo Fisher Scientific (2 mentions) Fixable viability dye efluor 450, by Thermo Fisher Scientific (2 mentions) Lsrii cytometer, by BD (2 mentions) Allophycocyanin conjugated annexin 5 and 7 aminoactinomycin d, by BD (2 mentions) D23844, by Thermo Fisher Scientific (2 mentions) Fitc annexin 5 apoptosis detection kit 1, by BD (1 mentions) Propidium iodide, by Merck Group (1 mentions) Rnase a, by Merck Group (1 mentions) Guava expresspro software, by Merck Group (1 mentions) Facscalibur, by BD (1 mentions)

Close spelling variants of this product

Guava EasyCyte flow cytometer (481 citations) Guava easyCyte (393 citations) Guava easyCyte HT (185 citations) Guava flow cytometer (112 citations) EasyCyte flow cytometer (44 citations) EasyCyte HT (11 citations) Guava easyCyte HT cytometer (10 citations) EasyCyte HT flow cytometer (10 citations) Guava® easyCyte HT Sampling Flow Cytometer (10 citations) Guava easyCyte HT 2-laser flow cytometer (2 citations)

63 protocols using guava easycyte ht flow cytometer

Radiation-Induced Apoptosis and Cell Cycle Analysis

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HCAEC were cultured on 6 cm² plates until confluent then irradiated with appropriate doses of IR. Cells were cultured for 7 days with media changed every other day. After 7 days, cells were rinsed twice with HBSS and tripsinised. At this point cells from each radiation dose were divided in half for apoptosis assay and cell cycle profiling. Apoptosis was measured by using FITC Annexin V Apoptosis Detection Kit I (BD Biosciences, cat. 556547) according to manufacturer’s protocol. Unstained control, FITC only and PI only controls were prepared for setting up the gates. The samples were analysed on Guava easyCyte HT flow cytometer (Merk Millipore) using apoptosis program.
Cells for cell cycle profiling were permeablised and fixed for 1 h in cold 70% ethanol. Following this, cells were centrifuged and re-suspended in 500 µl of HBSS containing 100 ng/µl of RNase A (Sigma-Aldrich) and 40 ng/µl of propidium iodide (Sigma-Aldrich) and incubated at 37°C for 30 min. The samples were analysed on Guava easyCyte HT flow cytometer (Merk Millipore) using cell cycle program.

Kabacik S, & Raj K. (2017). Ionising radiation increases permeability of endothelium through ADAM10-mediated cleavage of VE-cadherin. Oncotarget, 8(47), 82049-82063.

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Cell Cycle and Apoptosis Analysis of Renal Cancer Cells

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786-O and Caki-1 cells were seeded in 6 cm dishes and cultured in serum-free medium for 24 h before being treated with EVs for 24 h. Next, cells were harvested and fixed in 70% ethanol at 4°C for 2 h, and they were then incubated with RNase and the DNA-interacting dye propidium iodide (PI) for 30 min at room temperature. Cell cycle analyses were performed using a Guava easy Cyte HT flow cytometer (Merk, Germany).
For apoptosis analyses, 786-O and Caki-1 cells were seeded in 6-well plates and treated with EVs for 24 h. The cells were then collected, washed with PBS at 4°C, and resuspended in the 1 × binding buffer containing Annexin V-FITC and PI. After incubation for 15 min at room temperature, apoptosis was analyzed with a Guava easy Cyte HT flow cytometer (Merk, Germany). Cells in both the early and late stages of apoptosis were detected.

Meng L., Xing Z., Guo Z., Qiu Y, & Liu Z. (2021). Hypoxia-induced microRNA-155 overexpression in extracellular vesicles promotes renal cell carcinoma progression by targeting FOXO3. Aging (Albany NY), 13(7), 9613-9626.

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Flow Cytometry Analysis of Endothelial Cells

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For flow cytometry analysis of endothelial cells (ECs), cells were washed once in PBS, and enzymatically digested with 0.05% trypsin-EDTA (5 min at 37°C) and neutralized with knockout serum. Cells were centrifuged (300 x g for 5 min at room temperature) and resuspended in staining buffer (PBS + 0.05% BSA). Single cell suspensions (<1x10⁶ cells in 100 µL per tube) were incubated for 20 min on ice with 1µL of mouse anti-human CD31-APC (eBiosciences cat# 17-0319-42). Cells were washed with 3 mL of PBS, centrifuged (300 x g for 5 min at room temperature), and resuspended in 300 µL of staining buffer prior to acquisition. Viable cells were analyzed (1,000 events acquired for each sample) using the Guava EasyCyte HT flow cytometer and Guava ExpressPro software (Luminex) (EMD Millipore, Billerica, MA, USA) at the MSM’s RCMI core facility.

Thomas J.J., Harp K.O., Bashi A., Hood J.L., Botchway F., Wilson M.D., Thompson W.E., Stiles J.K, & Driss A. (2022). MiR-451a and let-7i-5p loaded extracellular vesicles attenuate heme-induced inflammation in hiPSC-derived endothelial cells. Frontiers in Immunology, 13, 1082414.

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Flow Cytometric Analysis of Cell Lines

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The OS cell lines were harvested and cell numbers were determined by utilizing the Countess Cell Counter (Invitrogen) followed by dilution in flow buffer (Dulbecco’s PBS with 0.5% BSA and 0.1% NaN₃). The cell suspension was distributed in a 96 well plate with 2 × 10⁵ cells per sample well. Primary antibodies were added in a concentration of 10 μg/ml and cells were incubated at 4°C for 30 min before three washes in 200 μl flow buffer. Secondary antibody, anti-mouse IgG F(ab’)2 fragment-FITC (Sigma-Aldrich, St Louis, MO, USA) or anti-human IgG-FITC (US Biologicals, Salem, MA, USA), was added and incubated for 30 min and washed as in the previous step. All wash steps were performed by centrifugation at 1,200 rpm for 5 min.
Washed cell pellets were dissolved in flow buffer and analyzed in a FACS Calibur (BD Bioscience, Franklin Lakes, NJ, USA) or Guava EasyCyte HT flow cytometer (Merck Millipore, Darmstadt, Germany). The flow analysis was replicated and reproduced at least three times for all cell lines. The data were analyzed with Kaluza Analysis 1.3 software (Beckman Coulter, Brea, CA, USA).

Westrøm S., Bønsdorff T.B., Abbas N., Bruland Ø.S., Jonasdottir T.J., Mælandsmo G.M, & Larsen R.H. (2016). Evaluation of CD146 as Target for Radioimmunotherapy against Osteosarcoma. PLoS ONE, 11(10), e0165382.

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Endothelial Cell Surface Phenotyping

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The surface phenotype of EA.hy926 endothelial cells was determined by flow cytometry. Cells were fixed as described above. Direct labelling was performed for the following markers: endoglin/CD105 (BD Biosciences, Le Pont de Claix, France), ICAM-1/CD54 (BD Biosciences, Le Pont de Claix, France), and Platelet endothelial cell adhesion molecule PECAM1/CD31 (BD Biosciences, Le Pont de Claix, France). The cells were incubated for 30 minutes with antibodies. In all experiments, background labelling was assessed using the relevant fluorochrome-conjugated mouse IgG isotype control (BD Biosciences, Le Pont de Claix, France). Cells were centrifuged at 300 g for 5 minutes at 22°C then washed in PBS. Data acquisition was performed using a Guava easyCyte HT Flow Cytometer (Merck Millipore, Molsheim, France) and analysis carried out using the Incyte program. At least 10,000 events were collected for each sample.

Cognasse F., Hamzeh-Cognasse H., Duchez A.C., Shurko N., Eyraud M.A., Arthaud C.A., Prier A., Heestermans M., Hequet O., Bonneaudeau B., Rochette-Eribon S., Teyssier F., Barlet-Excoffier V., Chavarin P., Legrand D., Richard P., Morel P., Mooney N, & Tiberghien P. (2022). Inflammatory profile of convalescent plasma to treat COVID: Impact of amotosalen/UVA pathogen reduction technology. Frontiers in Immunology, 13, 1034379.

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Pluripotency Marker Analysis in Cell Aggregates

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For flow cytometric analyses, the cultured aggregates were dissociated into single cells with TrypLE select and fixed with 4% PFA for 20 min at room temperature. Fixed cells were washed with phosphate buffered saline (PBS) containing 3% fetal bovine serum (FBS). To permeabilize for staining to intracellular protein markers, cells were treated in methanol on ice overnight. After blocking with 3% FBS/PBS, the cells were stained with antibodies; anti-OCT4-PE (653704, Biolegend, USA), and anti-Nanog-Alexa Fluor® 647 (674010, Biolegend, USA). These samples were compared to the samples of cells stained with isotype controls; mouse IgG2b-PE (400314, Biolegend, USA), mouse IgG1-Alexa Fluor® 647 (400136, Biolegend, USA). Flow cytometry data were acquired using a Guava® EasyCyte HT flow cytometer (Merck Millipore, USA) and analyzed using Guava®-InCyte software (Merck Millipore, USA).

Ibuki M., Horiguchi I, & Sakai Y. (2019). A novel tool for suspension culture of human induced pluripotent stem cells: Lysophospholipids as a cell aggregation regulator. Regenerative Therapy, 12, 74-82.

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Quantifying Macrophage Surface Markers

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Macrophages were first incubated with rat antimouse CD16/32 antibody (as the mouse Fc-gamma blocker) and then stained with FITC antimouse/human CD11b and goat antimouse SR-AI (detected with AlexaFluor 647 chicken antigoat secondary antibody) in 1% BSA and 0.1% sodium azide in 1× PBS. Cells were resuspended at ~0.5 million/mL and 20,000 events were acquired with Guava EasyCyte HT flow cytometer (Merck KGaA). The data were analyzed and plotted using FlowJo version 10.

Wang G., Serkova N.J., Groman E.V., Scheinman R.I, & Simberg D. (2019). Feraheme (Ferumoxytol) Is Recognized by Proinflammatory and Anti-inflammatory Macrophages via Scavenger Receptor Type AI/II. Molecular pharmaceutics, 16(10), 4274-4281.

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Cell Cycle and Apoptosis Analysis

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Before analysis of cell cycle distribution, cells were fixed in ice-cold 70% ethanol for 2 hours. After washing with PBS, cells were incubated with RNase (KeyGen Biotech Co. Ltd., Nanjing, China) for 30 min at 37°C followed by incubation with propidium iodide (PI) (KeyGen Biotech) for 30 min at 4°C. The results were examined on a Guava easyCyte HT flow cytometer (Merck-Millipore, Darmstadt, Germany) and analyzed with InCyte 2.7 software (Millipore). For analysis of apoptosis, cells were stained with Annexin V and 7-AAD (both from KeyGen Biotech) according to the manufacturer's instructions. Apoptotic fractions were analyzed using a FACScan cytometer (Becton-Dickinson, Franklin Lakes, NJ, USA). In our studies, the early apoptotic cells (Q4: Annexin V+/7-AAD-staining) and the late apoptotic cells (Q2: Annexin V+/7-AAD+staining) were considered to be undergoing apoptosis, and the numbers of these apoptotic cells as a proportion of total cells were analyzed.

Li X., Wang L., Su Q., Ye L., Zhou X., Song D, & Huang D. (2020). Highly Proliferative Immortalized Human Dental Pulp Cells Retain the Odontogenic Phenotype when Combined with a Beta-Tricalcium Phosphate Scaffold and BMP2. Stem Cells International, 2020, 4534128.

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Flow Cytometry Analysis of 4T1 Tumor Cells

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A single-cell suspension of tumor tissue was prepared for flow cytometry analysis, as previously described.^{52 (link)} Single cell isolates were preblocked with Ultra-LEAF purified antimouse CD16/32 antibody (BioLegend, San Diego, CA) in PBS supplemented with 2% FBS 10 min at 4 °C before antibody staining. The 4T1 single-cell suspension was stained with anti-F4/80-AF488. After incubation on ice for 20 min and washing, cells were analyzed for staining using the cells were analyzed with Guava EasyCyte HT flow cytometer (Merck KGaA). Cells were resuspended at ~0.5 million/mL and 20 000 events were detected. Dead cells and microparticles were excluded with FSC/SSC and the percentage of Cy5+ cells was analyzed with FlowJo software Version 10.6 (FlowJo, LLC). For immunostaining, mice with GL261 tumors were injected with DiI-amine liposomes and perfused 48 h postinjection. Brains were snap-frozen in OCT in liquid nitrogen and sectioned with a cryostat into 5–8 μm sections. The sections were fixed with 4% formalin on a slide, blocked with 10% goat serum, and stained with anti-CD11b and CD45 antibodies and corresponding secondary antibodies.

Wang G., Zannikou M., Lofchy L., Li Y., Gaikwad H., Balyasnikova I.V, & Simberg D. (2021). Liposomal Extravasation and Accumulation in Tumors as Studied by Fluorescence Microscopy and Imaging Depend on the Fluorescent Label. ACS nano, 15(7), 11880-11890.

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Quantifying Peptide Uptake in HeLa Cells

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HeLa cells were seeded onto 24-well plates (Sarstedt, Nümbrecht, Germany, 100,000 cells per well, respectively). After reaching 80–90% confluency, medium was removed, and cells were incubated with 1 µM CF-labeled peptide solution in serum-free medium for either 30 min or 2 h at 37 °C. The peptide solution was removed, cells were washed twice with PBS, and trypsinized for 5 min. Detached cells were resuspended in full medium (phenol red free) and afterwards analyzed using a guava easycyte HT flow cytometer (Merck, Darmstadt, Germany).

Lützenburg T., Burdina N., Scholz M.S, & Neundorf I. (2021). Improving Membrane Activity and Cargo Delivery Efficacy of a Cell-Penetrating Peptide by Loading with Carboranes. Pharmaceutics, 13(12), 2075.

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