MDA-MB-231 cells were cultured
in cell culture dishes at 37 °C using RPMI 1640 growth medium
with 10% FBS, 100 units per mL penicillin, and 100 μg/mL streptomycin.
The cells were then incubated in a humidified incubator at 37 °C
with 5% CO2. All of the cells were incubated at approximately
70% confluence until normal morphology was achieved. Then, the mixture
of MTX-CDs (100 μg/mL) in RPMI 1640 medium was added to each
dish. The cell dish was placed in an incubator for the desired time.
The cells were washed three times with 0.1 M phosphate-buffered saline
(PBS) (pH 7.4) to remove unattached compounds. For epifluorescence
and fluorescence imaging, a Nikon E1000M (Nikon, Tokyo, Japan) research
fluorescence microscope equipped with the Plan Apo apochromatic objectives
(Nikon, Tokyo, Japan) was used. The best fluorescence excitation was
observed when mirror cube units for 480–510 and 510–550
nm and a fluorescence microscope (Olympus microscope Bh2-FCA, Japan)
were used.
in cell culture dishes at 37 °C using RPMI 1640 growth medium
with 10% FBS, 100 units per mL penicillin, and 100 μg/mL streptomycin.
The cells were then incubated in a humidified incubator at 37 °C
with 5% CO2. All of the cells were incubated at approximately
70% confluence until normal morphology was achieved. Then, the mixture
of MTX-CDs (100 μg/mL) in RPMI 1640 medium was added to each
dish. The cell dish was placed in an incubator for the desired time.
The cells were washed three times with 0.1 M phosphate-buffered saline
(PBS) (pH 7.4) to remove unattached compounds. For epifluorescence
and fluorescence imaging, a Nikon E1000M (Nikon, Tokyo, Japan) research
fluorescence microscope equipped with the Plan Apo apochromatic objectives
(Nikon, Tokyo, Japan) was used. The best fluorescence excitation was
observed when mirror cube units for 480–510 and 510–550
nm and a fluorescence microscope (Olympus microscope Bh2-FCA, Japan)
were used.