A calcein-AM/propidium iodide (PI) double staining kit (Dojindo Molecular Technologies, Kumamoto, Japan) was used to assess the biocompatibility of prepared scaffolds according to the manufacturer’s instructions. Scaffolds carrying ADSCs were placed in 24-well plates, and then DMEM-F12 medium was added for culture under standard conditions at 37°C. On days 0, 3, and 7, scaffolds were collected, rinsed with PBS three times, and stained in 2-mL solution containing 2 μmol/L calcein-AM and 4.5 μmol/L PI at 37°C for 15 min. Fluorescent images of stained cells within scaffolds were obtained using a confocal laser scanning microscope (Leica TCS SP8, Leica, Germany). To evaluate ADSC viability at different time points, the numbers of live and dead cells were counted in the fluorescent confocal microscopy images using Image-Pro-Plus 6.0 software.
Calcein am propidium iodide double staining kit
Manufactured by Dojindo Laboratories
Sourced in Japan
The Calcein-AM/propidium iodide (PI) double staining kit is a laboratory tool used to simultaneously detect live and dead cells. Calcein-AM is a fluorescent dye that stains viable cells, while propidium iodide (PI) stains the nuclei of dead cells. This kit provides a simple and effective method for distinguishing between live and dead cells in a sample.
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2 protocols using calcein am propidium iodide double staining kit
1
Assessing ADSC Scaffold Biocompatibility
2
Cytotoxicity Evaluation of Cu12Sb4S13 Nanoparticles
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Huh7 and HCCLM3 human HCC cells were supplied by the Institute of Chinese Academy of Science, China. HepG2 cells were supplied by the American Type Culture Collection (Manassas, VA, USA). All these cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum and 1% antibiotics at 37 °C in 5% CO2. Cu12Sb4S13 nanoparticles were kindly provided by Chongshen Guo from the Key Laboratory of Microsystem and Microstructure in the Harbin Institute of Technology. mPEG5000-PLGA (MW: 15 kDa) was purchased from Jinan Daigang Bio-Technology, Inc. (Jinan, China). Perfluoropentane (PFP) and poly(vinyl alcohol) (PVA, 99% MW = 30,000–70,000 Da) were obtained from Sigma–Aldrich (St. Louis, MO, USA). Chloroform (CHCl3) and isopropyl alcohol were obtained from Chongqing East Chemical Industry Ltd., Co. (Chongqing, China). A Calcein-AM/ propidium iodide (PI) double staining kit was purchased from Dojindo Laboratories (Kumamoto, Japan). PD98059 was obtained from MCE (New Jersey, USA).
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