The viability of the cellular systems was evaluated by flow cytofluorimetry. After calibration and adjustment of negative control settings, analysis was performed using GuavaViaCount (Guava ViaCount Reagent, Millipore, Burlington, AS, USA) according to the standard color protocol and analysis according to the manufacturer’s recommendations. We used the flow cytofluorimeter MACSQuant Analyzer 7 (Miltenyi Biotec, Bergisch Gladbach, Germany).
Guava viacount
Manufactured by Merck Group
Sourced in United States
The Guava ViaCount is a cell counting and viability analysis instrument developed by Merck Group. It utilizes flow cytometry technology to provide accurate and reliable cell counts and viability measurements. The Guava ViaCount is designed for use in a variety of research and clinical applications.
Automatically generated - may contain errors
Lab products found in correlation
Axiovert s100 fluorescence microscope, by Zeiss (2 mentions)
Ficoll histopaque, by GE Healthcare (2 mentions)
Accuri c6, by BD (2 mentions)
Edta coated vacutainer tubes, by BD (2 mentions)
Ficoll paque, by GE Healthcare (2 mentions)
Collagenase type 2, by Merck Group (2 mentions)
Calcein am assay, by Bio-Techne (2 mentions)
Stempro accutase, by Thermo Fisher Scientific (1 mentions)
Guava easycyte 5ht flow cytometer, by Merck Group (1 mentions)
Ripa lysis buffer, by Beyotime (1 mentions)
Apo one homogeneous caspase 3 7 assay kit, by Promega (1 mentions)
Anti β actin, by Cell Signaling Technology (1 mentions)
Bca protein assay kit, by Beyotime (1 mentions)
Fluorescence based tubulin polymerization assay kit, by Cytoskeleton (1 mentions)
Wst 1, by Beyotime (1 mentions)
Guava flow cytometer, by Merck Group (1 mentions)
Live dead fixable aqua dead cell stain kit, by Thermo Fisher Scientific (1 mentions)
Countess automated cell counter, by Thermo Fisher Scientific (1 mentions)
Guava nexin reagent, by Merck Group (1 mentions)
Guava easycyte, by Merck Group (1 mentions)
31 protocols using guava viacount
1
Cell Viability Evaluation by Flow Cytometry
2
Cell Viability Assay using Guava
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Viability of cells was detected using Guava ViaCount (Merck Millipore, MA, USA). Briefly, the cells were seeded at density of 5 × 104 cells per well in a 12-well plate. Following overnight serum starvation, the cells were treated with LPA for indicated concentration and time periods. At the end of the treatment, cells were washed with PBS, trypsinized, and equal amounts of serum containing medium was added. 50 µL of cell suspension was stained with 150 µL of Guava ViaCount reagent, incubated for 5 min at room temperature and measured using Guava easyCyte 8 Benchtop Flow Cytometer (Merck Millipore). Total viable and dead cells were analyzed.
3
Cell Viability and Apoptosis Quantification
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Cell viability was quantified by Trypan Blue dye exclusion. Apoptotic nuclei were visualized and quantified by Hoechst 33342 dye using an Axiovert S100 Flfluorescence Microscope (Zeiss) equipped with a 40× oil objective (Zeiss) with excitation/emission at 350/535 nm. OCI-AML3 cells were subjected to Guava ViaCount (Millipore Sigma) to assess viable, dead, and apoptotic cell fractions and analyzed by flow cytometry. WEHI7.2 cells were subjected to dual staining with annexin-V and propidium iodide as previously described (4 (link)).
4
Cell Viability and Apoptosis Quantification
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Cell viability was quantified by Trypan Blue dye exclusion. Apoptotic nuclei were visualized and quantified by Hoechst 33342 dye using an Axiovert S100 Fluorescence Microscope (Zeiss) equipped with a 40× oil objective (Zeiss) with excitation/emission at 350/535 nm. OCI-AML3 cells were subjected to Guava ViaCount (Millipore Sigma) to assess viable, dead, and apoptotic cell fractions and analyzed by flow cytometry. WEHI7.2 cells were subjected to dual staining with annexin-V and propidium iodide as previously described [4 (link)].
5
Cell Viability and Apoptosis Quantification
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Cell viability was quantified by Trypan Blue dye exclusion. Apoptotic nuclei were visualized and quantified by Hoechst 33342 dye using an Axiovert S100 Fluorescence Microscope (Zeiss) equipped with a 40× oil objective (Zeiss) with excitation/emission at 350/535 nm. OCI-AML3 cells were subjected to Guava ViaCount (Millipore Sigma) to assess viable, dead, and apoptotic cell fractions and analyzed by flow cytometry. WEHI7.2 cells were subjected to dual staining with annexin-V and propidium iodide as previously described [4 (link)].
6
Cytotoxicity of Moringa oleifera Extract
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5 × 104 Huh7 cells were seeded in 6-well plates. After 24 h of culture, the first dose of aqueous M. oleifera extract (0, 30, 45, 60, 120, 250, and 500 μg/mL) was added and repeated two times every 24 h until cell viability was determined at 72 h with a Guava ViaCount (Guava Technologies) flow cytometer. The cell concentration of stained cells was between 1 × 104 and 5 × 105 cells/mL in accordance with the recommendations of the manufacturer.
7
Cell Viability Assay Protocol
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Cell death was assessed by the Guava ViaCount® assay (Guava Technologies, Millipore Corp.), according to the manufacturer’s instructions. After incubation of SCFAs in self-renewal conditions, the cell-culture medium and adherent cells previously detached with StemPro Accutase (A11105-01; Gibco™) were collected and centrifuged for 5 min at 600g and re-suspended in PBS with 2% foetal bovine serum. Cell suspension was mixed with Guava ViaCount reagent and incubated for 20 min at RT. Sample acquisition was performed using Guava easyCyte 5HT flow cytometer (Guava Technologies).
8
Influence of M2 Macrophages on CLL Cell Survival
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Example 258
Frozen purified CLL cells were obtained from a commercial vendor (All Cells). Macrophages were polarized into M0 (MCSF) or M2 (MCSF, IL4 for 2 days). M0 or M2 polarized macrophages were plated into a 96 well tissue culture plate at the stated density in RPMI+10% FCS with their respective polarization factors. The CLL cells were plated in the tissue culture plate at 150 thousand cells per well with media or M0 or M2 macrophages. CLL cell viability over time was measured by Guava viacount (Millipore). Co-cultures of M2 macrophages with CLL cells led to extended CLL cell survival (
The results indicate that PI3K-γ selective compounds can be used to treat CLL by potently inhibiting M2 activation and reducing survival of CLL cells.
9
Pediatric Blood Sample Collection
10
PBMC and Plasma Isolation from Blood
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