Copper source, cobalt source, and thiourea were purchased from Shanghai Aladdin Biochemical Technology Co., Ltd. (Shanghai, China). The Raw264.7 cells were obtained from the ScienCell Research Laboratories (California, USA). Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), penicillin and streptomycin were purchased from Gibco (New York, NY, USA). Anti-CD68 and anti-α-SMA antibodies were obtained from the Abcam company (Cambridge, UK). 6-dimercapto-2-phenylindole (DAPI) was purchased from DAKO (Santa Clara, CA, USA). CCK-8 and Calcein AM/PI reagents were purchased from Sigma-Aldrich (Shanghai, China) Trading Co., Ltd.
Calcein am pi
Manufactured by Merck Group
Sourced in United States
Calcein-AM/PI is a fluorescent dye kit used for live-cell and cell viability analysis. Calcein-AM is a cell-permeant dye that is converted to a green fluorescent product by active cells, while propidium iodide (PI) is a red fluorescent dye that can only enter cells with compromised membranes. This combination allows for the simultaneous detection of live and dead cells in a sample.
Automatically generated - may contain errors
4 protocols using calcein am pi
1
Copper-Cobalt-Thiourea Cytotoxicity Assay
2
Cell Viability Assay Using Calcein AM/PI
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Serum-starved cells were digested with trypsin-EDTA, and the cells were collected in centrifuge tubes and centrifuged at 1000 rpm for 3 min. The supernatant was removed, and the cell suspension was adjusted to 105-106 cells/ml with PBS and then mixed gently. This process was repeated several times to completely remove the medium. Then, 200 μl of the cell suspension was incubated at 37°C for 15 min after adding 100 μl of staining solution (calcein AM/PI) (Sigma-Aldrich, USA). Yellow-green living cells and red dead cells were observed at an excitation wavelength of 490 nm and 545 nm.
3
Multifunctional Nanoparticle for Biomedical Applications
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Sodium hyaluronate (MW = 1 000 000–150 000), 1,4-butaneglycol diglycidyl ether (BDDE), normal alkane, isobutanol, and Span 80 were purchased from Aladdin Technology Co., Ltd. Oleylamine, oleic acid, and thioacetamide provided by Heowns Biochemical Technology Co., Ltd. The polymethyl methacrylate (PMMA) microfluidic chip (the width and depth of the channel at the intersection are both 250 μm) was purchased from Beijing LiShi Micro-Nano Technology Co., Ltd. Bismuth neodecanoate was obtained from Shanghai Eon Chemical Technology Co., Ltd. Cell Counting Kit-8 (CCK-8), pancreatic digest, and penicillin/streptomycin was supplied by Beyotime Biotechnology Co., Ltd. Calcein-AM/PI was purchased from Sigma-Aldrich. All the rabbit blood samples were bought from Henan Yuechi Biotechnology Co., Ltd, and its hemolysis assay and blood coagulation test procedures were performed in accordance with the guidelines of Zhejiang Sci-Tech University.
4
Hydrogel Viability Assay of MC3T3-E1 Cells
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Fluorescent inverted microscopy was used to study the viability and morphology of MC3T3-E1 on the hydrogels using live/dead staining kit Calcein-AM/Propidium iodide (Calcein-AM/PI, Sigma, USA). Cells were cultured on the gels for 1 d and 5 d, then culture medium was removed and cell-gel constructs in each well were washed twice with PBS. Onemilliliter dye (10 μg/mL in PBS) was added into each well. After incubation for 45 min, the dye was removed and the constructs were washed once with PBS. The samples were observed with fluorescence inverted microscope (IX 71, Olympus, Japan) under blue fluorescent light (490 nm) and green fluorescent light (545 nm), respectively. Live cells were stained green and nucleus of dead cells were colored red.
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