Cellular apoptosis assay was performed with the Muse Annexin V and Dead Cell Kit (Merck Millipore) according to the manufacturer’s protocol as previously described [24 (link),38 (link)]. Apoptotic cells were detected and quantified using Muse Cell Analyzer flow cytometry (Merck Millipore).
Muse cell analyzer flow cytometry
Manufactured by Merck Group
Sourced in United States, Germany
The Muse Cell Analyzer is a flow cytometry instrument designed for automated cell analysis. It utilizes core flow cytometry technology to provide precise quantification of various cell characteristics, including cell count, viability, and subpopulations. The Muse Cell Analyzer is capable of analyzing a wide range of cell types and is suitable for use in a variety of research and clinical applications.
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5 protocols using muse cell analyzer flow cytometry
1
Quantification of Cellular Apoptosis
2
Annexin V Apoptosis Assay
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Cells were seeded on 6-well overnight and then treated with different concentrations of AA for 24 h. Cells were harvested and suspended in PBS (2% (v/v) BSA), and then incubated with Muse™ Annexin V & Dead Cell reagent (EMD Millipore, Billerica, MA, USA) for 20 min at room temperature in dark. Samples were analyzed by Muse Cell Analyzer flow cytometry (EMD Millipore, Billerica, MA, USA) and by MUSE 1.4 Analysis software (EMD Millipore).
3
Quantifying Cellular Oxidative Stress
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The user guide of the Muse Oxidative Stress Kit (Cat. No. MCH100111, Merck Millipore) describes the oxidative stress detection method. First, the cells were planted in a 6-well plate (1×104 cells/well) and incubated with various DC concentrations for 24 h as previously described (23 (link)). The collected cells were processed under conditions previously studied (23 (link)). The obtained cells were added to the Muse Oxidative Stress working solution reagent reaction at 37°C for 30 min, and the results were analyzed using Muse Cell Analyzer flow cytometry and the data were analyzed using the Muse Cell Soft V1.4.0.0 Analyzer Assays (Merck Millipore).
4
Cell Cycle and DNA Damage Analysis
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Cells were seeded on 60 mm dishes and incubated overnight. The next day, cells were treated with 40 μM SFN, 10 μM PEITC or 20 μM etoposide for desired time. Both floating and attached cells were collected and washed with ice-cold PBS, proceeded with the MuseTM Cell-Cycle Kit or MuseTM H2A.X Activation Dual Detection Kit (Merck Millipore, Germany) according to manufacturer’s instruction, and analyzed using Muse™ Cell Analyzer flow cytometry. 5000 counts were measured per sample.
5
Cell Viability Assay for Drug Screening
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Cell viability was determined following methods described previously (22 (link)). The cells (1×104 cells/well) were cultured in each well for 12 h and further treated with various DC concentrations for 24 h. These cells were collected and suspended in phosphate-buffered saline (PBS) followed by incubation with reagents contained in the Muse Annexin V and Dead Cell Kit (cat. no. MCH100105; Merck Millipore) in the dark at room temperature. Results were analyzed using Muse Cell Analyzer flow cytometry (Merck Millipore) and the data were analyzed using Muse Cell Soft V1.4.0.0 Analyzer Assays (Merck Millipore).
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