Guava easycyte ht cytometer

Manufactured by Merck Group

Sourced in United States

The Guava easyCyte HT cytometer is a compact and automated flow cytometry system designed for routine cell analysis. It provides high-throughput cell counting and analysis capabilities within a small footprint. The system utilizes a blue laser and up to four detectors to measure various parameters of cells, including size, granularity, and fluorescence intensity.

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Lab products found in correlation

Alexa fluor 647 conjugated goat anti rabbit antibody, by Thermo Fisher Scientific (1 mentions) Goat anti human mouse gremlin antibody, by R&D Systems (1 mentions) Ab20921, by Abcam (1 mentions) Cytofix cytoperm kit, by BD (1 mentions) Mab3582, by R&D Systems (1 mentions) Qfikit, by Agilent Technologies (1 mentions) Annexin 5 apoptosis detection kit apc, by Thermo Fisher Scientific (1 mentions) Annexin 5 apc, by Thermo Fisher Scientific (1 mentions) Synergy 2 multi mode reader, by Agilent Technologies (1 mentions) Caspase glo, by Promega (1 mentions) Hema 3, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Guava easyCyte (393 citations) Guava easyCyte HT (185 citations) Guava easyCyte HT flow cytometer (63 citations) EasyCyte HT (11 citations) Guava® easyCyte HT Sampling Flow Cytometer (10 citations) Guava easyCyte HT System Flow Cytometer (2 citations) Guava easyCyte HT 2-laser flow cytometer (2 citations)

10 protocols using guava easycyte ht cytometer

Spheroid Transfection and Flow Cytometry

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Six 2-day-old spheroids per well were transferred in non-treated 96-well plates. Forty-eight hours after transfection with fluorescently labeled SSOs ([ON705]-F complexed with JetMessenger and S₂₀-[ON705]-F), cells were trypsinized and resuspended in PBS. Single-cell suspensions were analyzed by flow cytometry on a Guava easyCyte HT Cytometer (Merck Millipore) using a 488-nm laser.

Nothisen M., Perche-Létuvée P., Behr J.P., Remy J.S, & Kotera M. (2018). Cationic Oligospermine-Oligonucleotide Conjugates Provide Carrier-free Splice Switching in Monolayer Cells and Spheroids. Molecular Therapy. Nucleic Acids, 13, 483-492.

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Quantifying Gremlin Expression in Cumulus Cells

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Following treatment with media or 5% obese follicular fluid for 24 hours, cumulus cells were fixed with 2% paraformaldehyde in a 96-well plate and washed with flow buffer containing phosphate-buffered saline (PBS), 0.1% bovine serum albumin, and 0.5% sodium azide. Saponin was used for cell permeabilization and immunostaining was performed with 25 μg/mL goat anti-human/mouse gremlin antibody (R&D Systems). Incubation with secondary antibody was performed using 4 μg/mL Alexa Fluor-647 conjugated rabbit anti-goat antibody (Invitrogen, Carlsbad, CA). Incubations were performed at 4°C for 30 minutes and cells were washed two times. Controls included negative and isotype matched antibodies. Samples and analyses were performed using the Guava easyCyte HT cytometer (EMD Millipore). The flow cytometry gating strategy is demonstrated in Supplemental Figure 1.

Kim T., Kim Y., Lucien F., Zhao Y, & Enninga E.A. (2020). Decreased gremlin 1 expression in women with BMI ≥35 kg/m2 is mediated by interleukin 10 and interleukin 1β in the follicular fluid. F&S science, 1(1), 16-26.

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Spore Fluorescence Analysis by Flow Cytometry

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At the indicated time after germination induction, samples of spores were washed in 30 mM EDTA and diluted to 500 cells per µl in YPD before analysis on Guava easyCyte HT cytometer (EMD Millipore). We recorded FCS and fluorescence signals in the orange emission spectrum (620/52 nm) after excitation with a violet laser (405 nm) of 5000 events.

Plante S, & Landry C.R. (2021). Closely related budding yeast species respond to different ecological signals for spore activation. Yeast (Chichester, England), 38(1).

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Spore Fluorescence Analysis by Flow Cytometry

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Plante S., & Landry C.R. (2020). Closely related budding yeast species respond to different ecological signals for spore activation.

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Flow Cytometric Analysis of IAV NP

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The following antibodies were used for flow cytometric staining: anti-IAV NP (Abcam, ab20921), Mouse IgG1 isotype control (Abcam, ab91356). Intracellular staining was performed using the BD Cytofix/Cytoperm kit (Becton Dickinson, 554714), according to the manufacturers instructions. All samples were run on a Guava easyCyte HT cytometer (Millipore) and analyzed using Flowjo (version 10.4.2, TreeStar)

Chamberlain N., Korwin-Mihavics B.R., Nakada E.M., Bruno S.R., Heppner D.E., Chapman D.G., Hoffman S.M., van der Vliet A., Suratt B.T., Dienz O., Alcorn J.F, & Anathy V. (2019). Lung epithelial protein disulfide isomerase A3 (PDIA3) plays an important role in influenza infection, inflammation, and airway mechanics. Redox Biology, 22, 101129.

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Quantification of Donor MHC Alloantigens

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The experiment was performed as reported previously.^{2 (link),9 (link)} Briefly, ipsilateral submandibular lymph nodes were harvested at 8 weeks after corneal transplantation. Cell suspension was blocked with Fc-blocking antibody (CD16/32; BD Bioscience, Franklin Lakes, NJ, USA) and stained for donor type major histocompatibility complex (MHC) class II I-A [b] alloantigen (BD Bioscience). Data were acquired and analyzed by a Guava easyCyte HT cytometer and InCyte 2.6 software (Millipore, Billerica, MA, USA).

Zhang L., Li G., Sessa R., Kang G.J., Shi M., Ge S., Gong A.J., Wen Y., Chintharlapalli S, & Chen L. (2017). Angiopoietin-2 Blockade Promotes Survival of Corneal Transplants. Investigative Ophthalmology & Visual Science, 58(1), 79-86.

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Quantifying Cell Surface Receptor Levels

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Example 9

Receptor surface expression levels on selected cell lines were determined using the QFIKIT (Dako K0078) employing flow cytometry, the results of which are shown in FIG. 18. Briefly, five populations of calibration beads presenting different numbers of mouse mAb molecules on their surfaces were used as a calibration standard. 1.5×105 cells/well were labeled with primary mouse anti-EGFR (ab187287, Abcam) and mouse anti-c-MET antibodies (MAB3582, R&D Systems) at saturating doses (5 μg/ml). Then, beads and cells were stained with secondary goat anti-mouse Fc F(ab′)2 FITC conjugate (10 μg/ml, Jackson Immuno Research) and were subjected to flow cytometry measurement using a Guava easyCyte HT cytometer (Millipore). Beads and cells were measured on the same day using the same settings. Based on a calibration line for fluorescence of beads versus bead surface density, antigen cell surface densities for c-MET and EGFR were calculated.

US11235063B2. Bi-specific antibodies for enhanced tumor selectivity and inhibition and uses thereof (2022-02-01). MERCK PATENT GMBH [DE]. Inventors: Achim Doerner [DE], Lars Toleikis [DE], Vanita D. Sood [US], Carolin Sellmann [DE], Christine Knuehl [DE].

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Cell Viability and Apoptosis Assays for K562 Cells

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For the cell viability assays, K562S or K562R cells were seeded in 96-well plates and treated as indicated prior to transient transfection with expression constructs. After 72 h, cell viability was measured using WST reagent (DoGenBio). For the apoptosis assays, K562S or K562R cells were seeded in 6-well plates and treated as indicated. After 72 h, cells were stained with Annexin V-APC and propidium iodide using an Annexin V-APC apoptosis detection kit (Thermo Fisher Scientific) and analyzed on a Guava EasyCyte HT cytometer (Millipore).

Yang T., Sim K.Y., Ko G.H., Ahn J.S., Kim H.J, & Park S.G. (2022). FAM167A is a key molecule to induce BCR-ABL-independent TKI resistance in CML via noncanonical NF-κB signaling activation. Journal of Experimental & Clinical Cancer Research : CR, 41, 82.

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Caspase-3/7 and Apoptosis Assay

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The cells were seeded in a 96-well plate with 10⁴ cells/well, with a blank control well without cells. Then, Caspase-Glo (G8091, Promega) was added to each well, shaken for 30 min and incubated for 2 h at room temperature. The Caspase-Glo 3/7 assay releases a substrate of luciferase after being cleaved by caspase-3 and -7. The substrate of luciferase results in a luminescent signal that can be detected by a Synergy 2 Multi-Mode Reader (BioTek).
Annexin V-APC (88-8007-72, eBioscience) was used for cell staining in the cell apoptosis assay. According to the manufacturer’s instructions, the cells (≥5 × 10⁵/well) were washed, digested and then resuspended using 200 µl 1× binding buffer. Then, 10 µl Annexin V-APC was added to 200 µl cell suspension and incubated in the dark at room temperature for 10–15 min. Cell apoptosis analysis was detected by a Guava easyCyte HT cytometer (Millipore).

Gao B., Zhao L., Wang F., Bai H., Li J., Li M., Hu X., Cao J, & Wang G. (2019). Knockdown of ISOC1 inhibits the proliferation and migration and induces the apoptosis of colon cancer cells through the AKT/GSK-3β pathway. Carcinogenesis, 41(8), 1123-1133.

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Bronchoalveolar Lavage Fluid Analysis

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BALF was collected by lavaging lungs with 1.0 mL of sterile PBS. Cells were isolated by centrifugation, and total cell counts were determined using a Guava easyCyte HT cytometer (Millipore) and analyzed using Flowjo (version 10.4.2, Ashland, OR: Becton, Dickinson and Company, Ashland, OR, USA). Differential cell counts were obtained via cytospins using Hema3-stained (Fisher Scientific) total cells, on a minimum of 300 cells/animal. The differential cell counts were normalized to the total cell counts [3 (link)].

Chamberlain N., Ruban M., Mark Z.F., Bruno S.R., Kumar A., Chandrasekaran R., Souza De Lima D., Antos D., Nakada E.M., Alcorn J.F, & Anathy V. (2022). Protein Disulfide Isomerase A3 Regulates Influenza Neuraminidase Activity and Influenza Burden in the Lung. International Journal of Molecular Sciences, 23(3), 1078.

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