Guava easycyte flowcytometry system

Manufactured by Merck Group

Sourced in United States

The Guava EasyCyte flow cytometry system is a compact and user-friendly instrument designed for cell analysis. It utilizes flow cytometry technology to measure and analyze various properties of cells, including size, granularity, and fluorescence. The system is capable of performing a range of cell-based assays and is suitable for applications in research, drug discovery, and clinical diagnostics.

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Lab products found in correlation

Fcs express software, by De Novo Software (9 mentions) Mitoprobe transition pore assay kit, by Thermo Fisher Scientific (4 mentions) Transdetect annexin 5 fitc pi cell apoptosis detection kit, by Transgene (3 mentions) Trypan blue exclusion assay, by Thermo Fisher Scientific (3 mentions) Methyl thiazolyl tetrazolium (mtt), by Merck Group (3 mentions) Annexin 5 and propidium iodide, by Thermo Fisher Scientific (3 mentions) Guava incyte software, by Merck Group (1 mentions) Lsm 700 microscope, by Zeiss (1 mentions) Guava cell cycle reagent, by Merck Group (1 mentions) Caspase inhibitor z vad fmk, by Selleck Chemicals (1 mentions) Crystal violet, by Thermo Fisher Scientific (1 mentions) Crystal violet, by Avantor (1 mentions) Annexin 5 propidium iodide, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Guava easyCyte (393 citations) Guava EasyCyte system (58 citations) Guava system (17 citations)

22 protocols using guava easycyte flowcytometry system

Quantifying Cellular ROS by Flow Cytometry

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For cellular
quantification of
ROS by flow cytometry, the cultures were grown in appropriate media
in 24-well culture plates up to 80% confluency. Culture media was
aspirated and replaced with 500 μL of stimulation media, containing
40 ng of TNF-α (Invitrogen Thermo, Carlsbad, US), and 100 ng
of IFN-γ (Gibco Thermo, Waltham, US) per 1 mL of the appropriate
cell medium. Drug treatments were administered and cells were incubated
for 24 h at 37 °C; nonstimulated and flow cytometry fluorophore-free
assay controls were run in parallel. Stimulation media was aspirated
and 300 μL of the appropriate cell medium lacking phenol red
and FBS, containing 1.25 μM of 2′,7′-dichlorodihydrofluorescein
diacetate (H₂DCFDA) (Thermo Scientific Waltham, USA) utilized
for the detection and bio-imaging of ROS, was added to each well.
The plate was incubated for 45 min at 37 °C, protected from light.
The cells were detached using 200 μL of trypsin and resuspended
with 300 μL of PBS containing 10% FBS, before centrifugation
at 1000g for 5 min. Pellets were resuspended in 500
μL of cold PBS and kept on ice. Cell sample flow cytometry was
performed on a Millipore Guava easyCyte flow cytometry system and
analyzed using the Guava InCyte software (Merck Millipore, Massachusetts,
US), measuring 5000 cells within set gating boundaries to calculate
a median fluorescent output.

Moore B.J., Islam B., Ward S., Jackson O., Armitage R., Blackburn J., Haider S, & McHugh P.C. (2019). Repurposing of Tranilast for Potential Neuropathic Pain Treatment by Inhibition of Sepiapterin Reductase in the BH4 Pathway. ACS Omega, 4(7), 11960-11972.

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Quantifying Lag-3 Receptor Expression and Ligand Binding

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Surface expression of the mouse Lag-3 receptor was measured by staining the 3T3-Lag-3 cells with 1.25 μg mL⁻¹ rat anti-mouse Lag-3 monoclonal antibody (eBioscience, Waltham, MA, USA) in phosphate-buffered saline (PBS) containing 0.05% (w/v) BSA on ice. After removal of unbound antibodies, cells were incubated with 3.75 μg mL⁻¹ FITC-conjugated goat anti-rat IgG Fc (Jackson Immunoresearch Laboratories) in PBS containing 0.05% (w/v) BSA on ice. After extensive washing, the surface fluorescence of the viable cells was measured on a guava easyCyte flow cytometry system (Merck, Kenilworth, NJ, USA). The Fgl1 binding activity of membrane-tethered Lag-3 receptor was determined by incubating the NIH-3T3 or NIH-3T3-Lag-3 cells with supernatant of pLNCX-Fgl1-mFc plasmid-transfected NIH-3T3 cells. After removal of the supernatant, cells were incubated with 3.75 μg mL⁻¹ FITC-conjugated goat anti-mouse IgG Fc (Jackson Immunoresearch Laboratories) in PBS containing 0.05% (w/v) BSA on ice. After removal of unbound antibodies by extensive washing in cold PBS containing 0.05% (w/v) BSA, the surface fluorescence of the viable cells was also measured on a guava easyCyte flow cytometry system.

Lin W.W., Ho K.W., Su H.H., Fang T.F., Tzou S.C., Chen I.J., Lu Y.C., Chang M.S., Tsai Y.C., Liu E.S., Su Y.C., Wang Y.T., Cheng T.L, & Huang H.K. (2021). Fibrinogen-Like Protein 1 Serves as an Anti-Inflammatory Agent for Collagen-Induced Arthritis Therapy in Mice. Frontiers in Immunology, 12, 767868.

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Isolation of Single-Cell Suspensions from Acr-Gfp Mice

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The cultured tissues of Acr‐Gfp transgenic mice were digested by a two‐step enzymatic procedure to generate single‐cell suspensions.13 They were first incubated in phosphate‐buffered saline (PBS) containing 1 mg/mL collagenase at 37°C for 15 minutes. After the centrifugation (190 g, 5 minutes), the supernatant was removed. Then, 0.25% trypsin solution was added and incubated at 37°C for 15 minutes. After the centrifugation and removal of the supernatant, cell pellets were resuspended in PBS supplemented with 5% FBS and stained by a final concentration of 2 μg/mL PI for several minutes. Samples were analyzed by the Guava® easyCyte flow cytometry system (Merck KGaA).

Komeya M., Yamanaka H., Sanjo H., Yao M., Nakamura H., Kimura H., Fujii T., Sato T, & Ogawa T. (2019). In vitro spermatogenesis in two‐dimensionally spread mouse testis tissues. Reproductive Medicine and Biology, 18(4), 362-369.

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Fluorescence-Based Analysis of Lipoplex Internalization

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Auto_shRFP@LS lipoplexes containing additional Cy5.5-labeled pT7/shRNA DNA fragments were transfected to B16F10 cells in 12-well plates at the final concentration of 15 μg/mL for 2 h, and the cells were washed three times with fresh PBS buffer. The cells were further incubated for 22 h and then detached from well plates by using trypsine-EDTA reagents. After the collected cells (2 × 10⁵ cells) were washed with DPBS including CaCl₂ and MgCl₂, the cells were sorted by Guava easyCyte flow cytometry system (Millipore, US) equipped with a red laser (excitation at 635 nm and emission at 665 nm). Additionally, confocal microscopic images were investigated by a Carl Zeiss LSM 700 microscope (Carl Zeiss, Germany) equipped with 514 nm laser for Cy5.5 fluorescence, while the images were processed and quantified by ZEN 2012 software. The nuclei were counterstained with DAPI.

Kwak S.Y., Han H.D, & Ahn H.J. (2019). A T7 autogene-based hybrid mRNA/DNA system for long-term shRNA expression in cytoplasm without inefficient nuclear entry. Scientific Reports, 9, 2993.

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Autophagy Analysis of SNO-HSA Treatment

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To investigate the effect of SNO-HSA on autophagy, an immunofluorescence study was carried out using an autophagy marker, microtubule-associated protein 1 light chain 3 LC3.29 (link) In a typical experiment, C26 cells were seeded in an 8-well imaging chamber (5 × 10⁴ cells/well) containing different concentrations of SNO-HSA, and the cells were incubated for 24 h. The cells were collected and fixed by treatment with 4% paraformaldehyde, following by a blocking process using 0.1% Triton X-100 in PBS (−) and then 1% BSA and 0.1% Tween 20 in PBS (−) (30 min each at room temperature). The cells were then treated with an anti-LC3 antibody for 2 h at room temperature according to the manufacturer's instructions. After treatment with a secondary antibody (anti-rabbit IgG–Alexa555) for 1 h at room temperature, the cells were washed with PBS (−) twice, resuspended in PBS (−), and seeded in an imaging chamber. The fluorescence was quantified using a Guava easyCyte Flow Cytometry system (Millipore, Bedford, MA, USA).
Details of other experimental procedures are given in Document S1.

Ishima Y., Inoue A., Fang J., Kinoshita R., Ikeda M., Watanabe H., Maeda H., Otagiri M, & Maruyama T. (2015). Poly-S-nitrosated human albumin enhances the antitumor and antimetastasis effect of bevacizumab, partly by inhibiting autophagy through the generation of nitric oxide. Cancer Science, 106(2), 194-200.

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Cell Cycle Analysis of Hsp90 Inhibitors

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1 × 10⁶ HepG2 cells were seeded in the six-well plates with 10% FBS DMEM. Cells were treated with Hsp90 inhibitors (STA9090/NB) or co-cultured with 10 μg of EVs (C/STA9090/NB-induced) for 24 h (5% CO₂, 37 °C). Then cells were harvested and immediately fixed with 75% ethanol at −20 °C (overnight). For the cell cycle analysis, cells were stained with PI (50 μg/mL, RN3501, Aidlab Biotechnologies Co Ltd, Beijing, China) and RNase (50 μg/mL, R30396–10×1 ml, YuanYe Bio-Technology Co Ltd, Shanghai, China) for 30 min separately, and cell cycle was detected by Guava easyCyte Flow Cytometry System (6HT2L, Millipore, MA, USA). The data were analyzed by the Modfit software 4.2 (BD Biosciences, Franklin Lake, New Jersey, USA).

Tan W., Zhang J., Liu L., Liang M., Li J., Deng Z., Zheng Z., Deng Y., Liu C., Li Y., Xie G., Zhang J., Zou F, & Chen X. (2022). Hsp90 Inhibitor STA9090 induced VPS35 related extracellular vesicle release and metastasis in hepatocellular carcinoma. Translational Oncology, 26, 101502.

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Cell Cycle and Apoptosis Analysis

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Cell cycle and apoptosis analyses were performed on a Guava EasyCyte Flow Cytometry System (Millipore, Billerica, MA, USA). Cells were seeded in 6-cm culture plates and treated with 2 μM TBPT or an equal volume of DMSO on the next day for the indicated times. To analyze the intracellular DNA content, cells were harvested, washed twice in PBS and fixed in 70% ethanol at −20 °C overnight. After the cells were washed in PBS, they were stained with Guava cell cycle reagent (Millipore) and analyzed via flow cytometry. For cell apoptosis analysis, the cells were harvested, washed in PBS, stained with PI/Annexin V-FITC reagent (Life Technologies) and analyzed by flow cytometry.

Mu Y., Liu Y., Xiang J., Zhang Q., Zhai S., Russo D.P., Zhu H., Bai X, & Yan B. (2016). From fighting depression to conquering tumors: a novel tricyclic thiazepine compound as a tubulin polymerization inhibitor. Cell Death & Disease, 7(3), e2143-.

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Quantifying Apoptosis in A431 Cells

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A431 cells were seeded on a 60-mm dish and transfected with siBAX_01, siBAX_02, or siNC oligos and cultured for 48 h. A TransDetect Annexin V-FITC/PI cell apoptosis detection kit (TransGen Biotech, Beijing, China) was applied according to the manufacturer’s protocols. Cell apoptosis was detected and quantified by Guava easyCyte Flow Cytometry System (Millipore, Billerica, MA, USA).

Zhou L., Gao R., Wang Y., Zhou M, & Ding Z. (2017). Loss of BAX by miR-365 Promotes Cutaneous Squamous Cell Carcinoma Progression by Suppressing Apoptosis. International Journal of Molecular Sciences, 18(6), 1157.

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Luteolin-Induced Apoptosis Analysis

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Cells were treated with luteolin for 24 h. After collection, the cells were washed in ice-cold PBS. Subsequently, the cells were centrifuged at 1,000 rpm for 10 min and washed twice with PBS. The cells were resuspended with a binding buffer (0.1 M Hepes/NaOH pH 7.4, 25 mM CaCl₂, 1.4 M NaCl), containing PI and Annexin V- FITC, and stored in the dark at room temperature for 20 min. Apoptosis of cells analyzed by Guava easyCyte flow cytometry system (Millipore Corporation, Billerica, MA, USA).

Woo J.H., Jang D.S, & Choi J.H. (2021). Luteolin Promotes Apoptosis of Endometriotic Cells and Inhibits the Alternative Activation of Endometriosis-Associated Macrophages. Biomolecules & Therapeutics, 29(6), 678-684.

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Apoptosis Quantification in A375 Cells

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A375 cells were seeded on a 60-mm dish and transfected with NC ASO, PURPL ASO1, or PURPL ASO2 and cultured for 48 h. TransDetect Annexin V-FITC/PI cell apoptosis detection kit (TransGen Biotech) was applied according to instructions. Cell death was detected and quantified using a Guava easyCyte Flow Cytometry System (Merk Millipore).

Han S., Li X., Wang K., Zhu D., Meng B., Liu J., Liang X., Jin Y., Liu X., Wen Q, & Zhou L. (2021). PURPL represses autophagic cell death to promote cutaneous melanoma by modulating ULK1 phosphorylation. Cell Death & Disease, 12(11), 1070.

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