Cell suspension was prepared in RPMI1640 medium supplemented with 10% FBS and antibiotics and exposed to nsPEFs as described above. At 6 h after nsPEF exposure, cell viability was analyzed using a CellTiter-Glo luminescent cell viability assay kit (Promega, WI, USA) according to the manufacturer’s procedures. Luminescence was measured using a 2030 ARVO X multilabel reader (Perkin Elmer).
2030 arvo x multilabel reader
Manufactured by PerkinElmer
Sourced in United States
The 2030 ARVO X multilabel reader is a laboratory instrument designed to detect and measure fluorescence, luminescence, and absorbance in a variety of microplate-based assays. It features a xenon flash lamp, precision optics, and a sensitive detector to provide accurate and reliable results.
Automatically generated - may contain errors
4 protocols using 2030 arvo x multilabel reader
1
nsPEF Exposure and Cell Viability
2
Cell Viability and ATP Measurement
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Cell viability was measured using a Cell Counting Kit-8 (Dojindo Laboratories, Japan) and a microplate reader (MPR-A100, AsOne, Osaka, Japan). Cell lysis and ATP measurement were carried out using a CellTiter-Glo 3D cell viability assay reagent (G9682, Promega, Madison, WI, USA) according to the manufacturer’s instructions. Luminescence was measured using a 2030 ARVO X multilabel reader (Perkin Elmer, Shelton, CT, USA).
3
ATP Depletion Assay in Cells
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Cells were plated in a 96-well plate (5 × 103 cells/well). After 24 h incubation at 37 °C, medium was removed, and fresh alphaMEM containing 25 mM 2DG and 10 µM CCCP was added to each well. Cell lysis and ATP measurement were performed using a CellTiter-Glo luminescent cell viability assay kit (Promega) according to the manufacturer’s instructions. Luminescence was measured using a 2030 ARVO X multilabel reader (Perkin Elmer, USA).
4
Quantification of Extracellular DNA
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For the measurement of extracellular DNA, cell suspension was treated with 0.1 unit/µl MNase and 1 µg/ml RNase A at room temperature for 5 min. The MNase reaction was stopped by adding EDTA at 10 mM, and the cells were removed by centrifugation at 200 × g for 2 min. SYTOX Green was added to the supernatant at 2.5 µM, and fluorescence was measured using a 2030 ARVO X multilabel reader (Perkin Elmer, MA, USA). For the measurement of total DNA, cells were suspended in HBS containing 0.5% Triton X-100 and lysed by three cycles of freeze-thaw. Cell lysates were reacted with 0.1 unit/µl MNase and 1 µg/ml RNase A at room temperature for 5 min. EDTA (10 mM) and SYTOX Green (2.5 µM) were added to the lysates, and fluorometric measurement was performed as described above. DNA extrusion was expressed as a ratio of fluorescence for extracellular DNA to that for total DNA. When Ca2+-free HBS was used (Fig. 6D ), CaCl2 solution was added to cell suspension prior to MNase treatment to yield 2 mM Ca2+, as MNase requires Ca2+ for its catalytic activity.
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