Intracellular nano-silver and silver ion quantitative determination method
A determination method and technology for silver ions are applied in the field of quantitative determination of nano-silver and silver ions, which can solve the problems of inability to quantitatively measure nano-silver and inability to distinguish nano-silver.
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Embodiment 1
[0063] cell culture
[0064] The zebrafish larval cells derived from one day old were used as the cells to be tested. Under the condition of 28°C, the DMEM medium supplemented with 10% fetal bovine serum was used as the culture medium to culture the cells to be tested, and subcultured to logarithmic Cells in the growth phase were used for subsequent experiments.
Embodiment 2
[0066] Determination of AIE Fluorescent Probe Concentration
[0067] Using Thiazolium Blue Colorimetric Method (MTT Method) to Investigate the Toxicity of AIE Fluorescent Probe
[0068] The cells to be tested in the logarithmic growth phase were collected, transferred to a 96-well plate at a density of 5000 cells / well, and cultured overnight at 28°C; the old medium was removed, and the cell pellet was collected. In the cell pellets, new culture medium containing different concentrations of AIE fluorescent probes were added, and cultured at 28°C for 6 hours; then 10 μL of MTT solution with a concentration of 5 mg / mL was added to each well, and the Incubate at 37°C for 4 hours; add 100 μL of a mixed solution consisting of 10% sodium lauryl sulfate and 0.01mol / L hydrochloric acid to each well, and incubate at 37°C for 6 hours to determine its Absorbance at 595nm, wherein, the zeroing sample is a mixture of culture medium, MTT, and dimethyl sulfoxide; the control sample is a mixt...
Embodiment 3
[0072] Determination of the concentration of silver ion fluorescent probe TEZ-TPE-1
[0073] Toxicity of Silver Ion Fluorescent Probe TEZ-TPE-1 Using Thiazolium Blue Colorimetric Method (MTT Method)
[0074] The cells to be tested in the logarithmic growth phase were collected, transferred to a 96-well plate at a density of 5000 cells / well, and cultured overnight at 28°C; the old medium was removed, and the cell pellet was collected. In the cell pellet, silver ion fluorescent probes containing different concentrations (0 μmol / L, 5 μmol / L, 10 μmol / L, 20 μmol / L, 30 μmol / L, 50 μmol / L, 70 μmol / L, 100 μmol / L) were added respectively. Incubate the new medium for TEZ-TPE-1 at 28°C for 6 hours; add 10 μL of MTT solution with a concentration of 5 mg / mL to each well, and incubate at 37°C for 4 hours; Add 100 μL of a mixed solution consisting of 10% sodium lauryl sulfate and 0.01 mol / L hydrochloric acid to each well, incubate at 37°C for 6 hours, and measure its absorbance at 595 nm, wh...
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