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Muse count viability assay kit

Manufactured by Merck Group
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The Muse® Count & Viability Assay Kit is a compact, automated cell analysis system that provides cell count and viability data. The kit utilizes fluorescent dyes and the Muse® Cell Analyzer to deliver quick, reliable results.

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Lab products found in correlation

Muse cell analyzer, by Merck Group (17 mentions) Muse cell analyser, by Merck Group (4 mentions) Hoechst 33342, by Thermo Fisher Scientific (3 mentions) Facscalibur flow cytometer, by BD (3 mentions) Penicillin, by Thermo Fisher Scientific (3 mentions) Streptomycin, by Thermo Fisher Scientific (3 mentions) Lipofectamine 3000, by Thermo Fisher Scientific (2 mentions) Annexin 5 dead cell kit, by Merck Group (2 mentions) Fetal calf serum (fcs), by Harvard Bioscience (2 mentions) Acetylated lysine, by Cell Signaling Technology (1 mentions) Suramin, by Merck Group (1 mentions) Cleaved and total caspase 3, by Cell Signaling Technology (1 mentions) Anti flag m2 magnetic bead, by Merck Group (1 mentions) Aicar, by Merck Group (1 mentions) Dithiothreitol (dtt), by Merck Group (1 mentions) Cell culture reagents, by GE Healthcare (1 mentions) N6022, by Merck Group (1 mentions) 3 4 5 dimethyl 2 thiazolyl 2 5 diphenyl 2 h tetrazolium bromide mtt, by Sangon (1 mentions) Attractene, by Thermo Fisher Scientific (1 mentions) Fluor de lys sirt1 fluorometric drug discovery assay kit, by Enzo Life Sciences (1 mentions)

Close spelling variants of this product

Muse Count and Viability Assay Kit Muse Count & Viability Kit MuseTM

32 protocols using muse count viability assay kit

1

Flow Cytometric Analysis of Cell Count, Viability, and Apoptosis

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To estimate the total cell count, cell viability, and apoptotic cell population, flow cytometric analysis was carried out using Muse® Count & Viability Assay Kit (Millipore, Billerica, MA, USA). In brief, B16F10 cells were plated at a density of 1 × 104 cells/mL overnight and treated with the indicated concentrations of flumequine (0–400 μM) for 72 h. The cells were harvested and washed with ice cold phosphate-buffered saline (PBS). Then, the cells were incubated with the Muse® Count & Viability Assay Kit for 5 min and analyzed according to the manufacturer’s instructions by Muse® cellcycler (Millipore). H2O2 (100 μM) was used as an apoptosis-inducing control.
Karunarathne W.A., Molagoda I.M., Kim M.S., Choi Y.H., Oren M., Park E.K, & Kim G.Y. (2019). Flumequine-Mediated Upregulation of p38 MAPK and JNK Results in Melanogenesis in B16F10 Cells and Zebrafish Larvae. Biomolecules, 9(10), 596.
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2

AMPK-p53 Signaling Modulation in Cell Assays

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Cell culture reagents were purchased from GE and fetal bovine serum (FBS) was purchased from PAN-Biotech; DTT was purchased from Millipore (Massachusetts, USA); Compound C, AICAR, Resveratrol, Suramin, SNAP, and ODQ were purchased from Sigma-Aldrich; N6022, Pifithrin-α, VO-Ohpic trihydrate, AS1842856, and MK 2206 were from MCE. Hydrogen peroxide (H2O2) was from Sinopharm Chemical Reagent; 3-(4,5-dimethyl-2-thiazolyl)−2,5-diphenyl-2-H-tetrazolium bromide (MTT) was purchased from Sangon Biotech; Lipofectamine® 3000 and Attractene were from Thermo Fisher Scientific and Qiagen, respectively. Fluor-de-Lys SIRT1 fluorometric drug discovery assay kit was from Enzo Life Sciences; Hydrogen Peroxide Assay Kit, Nitric Oxide Assay Kit, RIPA lysis buffer, Bicinchoninic Acid assay Kit, and Nuclear and Cytoplasmic Protein Extraction Kit were from Beyotime; Super ECL Prime was from US Everbright®Inc. The mRNA extraction kit was from Bioteke. Protein A Magnetic Beads and the Muse Count & Viability Assay Kit were purchased from Millipore. Anti-Flag M2 Magnetic Beads was from Sigma-Aldrich. Antibodies for phospho- and total-AMPK, acetyl- and total-p53, SIRT1, Cleaved- and total-Caspase-3, FOXO1, PTEN, β-actin, and acetylated-Lysine were from Cell Signaling Technology; antibodies for P21 was from Abcam; antibodies for Prdx2 was from Santa Cruz Biotechnology.
Zhang Y., Sun C., Xiao G., Shan H., Tang L., Yi Y., Yu W, & Gu Y. (2019). S-nitrosylation of the Peroxiredoxin-2 promotes S-nitrosoglutathione-mediated lung cancer cells apoptosis via AMPK-SIRT1 pathway. Cell Death & Disease, 10(5), 329.
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3

Comparative Viability Assays for ATRT Cell Lines

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Cells were plated on a 96-well plate, with 2000 cells/well for cell lines BT-12, BT-16, BT-37, CHLA-05-ATRT, and CHLA-06-ATRT. CHLA-02-ATRT and CHLA-04-ATRT were grown at a density of 200,000 cells/well in a 6-well plate, then plated in a 96-well plate. All cells were plated in triplicate in all conditions. Mitochondrial activity was determined with the MTS 3-(4, 5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) from Promega. 40μL of MTS reagent was added to 200μL of media containing cells and/or drugs per well, and incubated for 1 hour at 37°C with 5% CO2, then measured with Epoch Micro-Volume Spectrophotometer Plate Reader System (Biotek) at 490nm absorbance. Measurements were taken at Day 0, 3, and 5.
Cell viability assays were completed with MUSE Count & Viability Assay Kit (Millipore MCH100102). Cells were prepared in a uniform cell suspension for counting, and cells were incubated with Muse Count & Viability Reagent for 5 minutes. Cell viability was analyzed on Muse Cell Analyzer.
Wang S.Z., Poore B., Alt J., Price A., Allen S.J., Hanaford A.R., Kaur H., Orr B.A., Slusher B.S., Eberhart C.G., Raabe E.H, & Rubens J.A. (2019). Unbiased metabolic profiling predicts sensitivity of high MYC-expressing atypical teratoid/rhabdoid tumors to glutamine inhibition with 6-diazo-5-oxo-L-norleucine. Clinical cancer research : an official journal of the American Association for Cancer Research, 25(19), 5925-5936.
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4

Multimodal Cell Death Assay

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Cell death was measured using a Cell Meter Apoptotic and Necrotic Detection kit (ATT Bioquest, Sunnyvale, CA, USA) as previously described.24 (link) In brief, cells were incubated at 37 °C for 30 min with Apopxin Green for detection of phosphatidylserine on cell surface, PI or 7-ADD for labeling the nucleus of cells with membrane rupture, and CytoCalcein for labeling live cell cytoplasm. Cell death was then analyzed with an EVOS FL digital fluorescence microscope (AMG) or a FACSCalibur flow cytometer (BD Biosciences, San Jose, CA, USA). Cells with chromatin condensation were visualized by Hoechst 33342 (Invitrogen, Waltham, MA, USA) staining. Cell viability was also assessed using the Muse Count & Viability assay kit (Millipore, Billerica, MA, USA). In brief, cells were trypsinized, washed, and incubated with the Muse Count & Viability reagent, and cell viability was quantified on a Muse cell analyzer (Millipore).
Guo X., Yin H., Chen Y., Li L., Li J, & Liu Q. (2016). TAK1 regulates caspase 8 activation and necroptotic signaling via multiple cell death checkpoints. Cell Death & Disease, 7(9), e2381-.
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5

Glioblastoma Cell Line Characterization

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Human U87-MG glioblastoma cells (ATCC HTB-14) were modified by lentiviral infection to stably express fLuc2 and eGFP (U87-fLuc2/eGFP, maximum excitation λ = 488 nm, maximum emission λ = 509 nm) under the control of the CMV promoter (kindly provided by Dr. S.A. Collins from Department of Medicine-Digestive Diseases, UCLA, USA)48 (link). They were grown at 37 °C in a 5% CO2 humidified atmosphere, in Modified Essential Medium EARLES (10370–047, Gibco, France) supplemented with 10% fetal bovine serum (10500–064, Gibco, France), 0.2% Glucose (19002–013, Gibco, France), 2 mM L-glutamine (X0550, Dominique Dutscher, France) and 100 U/ml penicillin and 100 μg/ml streptomycin (15140155, Gibco, France). The viability of U87 cells was greater than 90% as determined by visual counts using Trypan Blue Dye Exclusion on a Malassez cell, or by analysis with the Muse® Cell Analyzer (MCH100102, Muse Count & Viability Assay Kit, Millipore).
Bardet S.M., Carr L., Soueid M., Arnaud-Cormos D., Leveque P, & O’Connor R.P. (2016). Multiphoton imaging reveals that nanosecond pulsed electric fields collapse tumor and normal vascular perfusion in human glioblastoma xenografts. Scientific Reports, 6, 34443.
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6

Preparation of Conditioned Medium from MSCs

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For the preparation of the conditioned medium (CM), MSCs (passage 3–7) were cultured in the presence of their expansion medium until 80% of confluence. Conditioned medium was obtained from supernatants of 8.95 ± 0.46 × 103/cm2 MSCs maintained in RPMI medium supplemented with 0.1% BSA for 16 h. The viability of MSCs after starvation was about 86 ± 0.5% as detected by the Muse® Count &Viability Assay Kit (CTRL normal medium 88,9% of vitality) (Millipore, MA, USA). Supernatant was first centrifuged at 1500 g for 20 min, to remove debris and apoptotic bodies and then concentrated at 4 °C, approximately 200-fold, using ultrafiltration units (Amicon Ultra-PL 3, Millipore) with a 3 kDa molecular weight cut-off as previous described [21 (link)]. After the concentration CM-containing EVs in 1% dimethyl sulfoxide was kept at −80 °C until use.
Collino F., Pomatto M., Bruno S., Lindoso R.S., Tapparo M., Sicheng W., Quesenberry P, & Camussi G. (2017). Exosome and Microvesicle-Enriched Fractions Isolated from Mesenchymal Stem Cells by Gradient Separation Showed Different Molecular Signatures and Functions on Renal Tubular Epithelial Cells. Stem Cell Reviews, 13(2), 226-243.
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7

Spleen Isolation and Cell Preparation

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Spleens were removed by incising the left-hand side of the abdomen, after cardiac bleed and cervical dislocation of the animal, and transferred to gentleMACs™ C tubes (Miltenyi Biotec) containing autoMACS® running buffer (Miltenyi Biotec). Spleens were macerated in a gentleMACS™ Dissociator (Miltenyi Biotec) using the mouse spleen pre-set program and then tubes were centrifuged at 250 × g for 10 min. The supernatants were discarded, and pellets resuspended in red blood cell lysis solution for 5 min. Sterile PBS was added to quench the lysis solution after 5 min and the cell suspensions were passed through 70 um filter into 50 mL Falcon tubes using Pasteur pipettes. The filtered solutions were centrifuged again at 250 ×g for 10 min, supernatants discarded, and pellets resuspend in 10 mL of complete DMEM. A Muse® Count & Viability Assay Kit (Millipore) was used to measure the final cell count and viability of each sample on a Muse® Cell Analyzer instrument (Millipore). Samples were diluted to 4 × 106 in complete DMEM.
Sheerin D., Dold C., Silva-Reyes L., Linder A., Pollard A.J, & Rollier C.S. (2022). Inclusion of a dual signal sequence enhances the immunogenicity of a novel viral vectored vaccine against the capsular group B meningococcus. Cell & Bioscience, 12, 86.
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8

Nicotine-Induced Oral Epithelial Cell Response

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Oral epithelial cells were treated with 150 mM nicotine hemisulphate salt solution (Sigma-Aldrich, MO) and incubated at 37 °C and 5 % CO2 for 10 days with the passage every 2 days. Untreated cells were also cultured under the same conditions except for nicotine at the same time. The media along with 150 mM nicotine was replaced every third day and were passaged at 90 % confluence in both treated and untreated plates. After 10 days of incubation, treated and untreated cells were collected, washed with PBS, and were proceeded further for cell count and viability assay (Muse® Count &Viability Assay kit) as per the manufacturer's protocol using Muse cell analyzer (Muse® Cell Analyzer, Millipore, USA).
Joshi J., Pandit A, & Shah F. (2023). Nicotine mediated epithelial modulations: An in-vitro evidence. Journal of Oral Biology and Craniofacial Research, 13(6), 796-800.
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9

Multiparametric Apoptosis and Viability Assay

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Cell death was measured using a Cell Meter Apoptotic and Necrotic Detection kit (ATT Bioquest, Sunnyvale, CA, USA) as previously described37 (link). Briefly, cells were incubated at 37 °C for 30 min with Apopxin Green for detection of phosphatidylserine on cell surface, PI or 7-ADD for labeling the nucleus of cells with membrane rupture, and CytoCalcein for labeling live cell cytoplasm. Cell death was then analyzed with an EVOS FL digital fluorescence microscope (AMG) or a FACSCalibur flow cytometer (BD Biosciences, San Jose, CA, USA). Cells with chromatin condensation were visualized by Hoechst 33342 (Invitrogen, Waltham, MA, USA) staining. Cell viability was also assessed using the Muse Count & Viability assay kit (Millipore, Billerica, MA, USA). In brief, cells were trypsinized, washed, and incubated with the Muse Count & Viability reagent, and cell viability was quantified on a Muse cell analyzer (Millipore).
Guo X., Hong S., He H., Zeng Y., Chen Y., Mo X., Li J., Li L., Steinmetz R, & Liu Q. (2020). NFκB promotes oxidative stress-induced necrosis and ischemia/reperfusion injury by inhibiting Nrf2-ARE pathway. Free radical biology & medicine, 159, 125-135.
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10

Cell Growth and Apoptosis Analysis

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The cell growth curve was determined by counting the cell number on particular days using Muse® Cell Analyzer (Millipore) and Muse® Count &Viability Assay Kit (Millipore). The level of apoptosis was detected 24 h after irradiation. Muse® Caspase-3/7 Assay Kit (Millipore) was used according to the manufacturer’s protocol and stained cells were measured by Muse® Cell Analyzer (Millipore).
Konířová J., Cupal L., Jarošová Š., Michaelidesová A., Vachelová J., Davídková M., Bartůněk P, & Zíková M. (2019). Differentiation Induction as a Response to Irradiation in Neural Stem Cells In Vitro. Cancers, 11(7), 913.
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