For in vitro experiments, the KB cells, treated with 15 μg/ml of TNHs and ZnO NCs and with the corresponding number ofEVs, were imaged after 24 h ofincubation at 37°C, 5% CO2. The cells were counted and seeded (3 × 104 cells/well) in 4-well chamber slides (Nunc™ Lab-Tek™ II CC2™ Chamber Slide System, Thermo Fisher Scientific™, MA, USA). After 24 h of growth in standard conditions, the medium was replaced with 500 μl of treatment solution containing the particle for a further 24 h. To maintain healthy cells, in their physiological condition during the acquisition time, an incubator gas chamber (Okolab) equipped with CO2 sensors, temperature unit and an active humidity controller was used. In order to label cell membranes, 2.5 μl of wheat germ agglutinin conjugated with Alexa Fluor 488 dye (λEx = 495 nm) was added to the cell samples and after 10 min of incubation at 37°C, 5% CO2, two washing steps were performed by using 500 μl of live-cell imaging solution (1×, Molecular Probes) at 37°C. ZnO NCs were used as labeled with Atto647 NHS ester dye, EVs were labeled in DiD and in DiO for sample treated with EVs and TNHs, respectively.
Incubator gas chamber
Manufactured by Okolab
The Incubator Gas Chamber is a specialized laboratory equipment designed to provide a controlled environment for cell culture experiments. It maintains precise control over temperature, humidity, and gas composition, ensuring optimal conditions for cell growth and observation.
Automatically generated - may contain errors
2 protocols using incubator gas chamber
1
Visualizing Cellular Interactions with TNHs and EVs
2
Visualizing Cellular Interactions with TNHs and EVs
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For in vitro experiments, the KB cells, treated with 15 μg/ml of TNHs and ZnO NCs and with the corresponding number ofEVs, were imaged after 24 h ofincubation at 37°C, 5% CO2. The cells were counted and seeded (3 × 104 cells/well) in 4-well chamber slides (Nunc™ Lab-Tek™ II CC2™ Chamber Slide System, Thermo Fisher Scientific™, MA, USA). After 24 h of growth in standard conditions, the medium was replaced with 500 μl of treatment solution containing the particle for a further 24 h. To maintain healthy cells, in their physiological condition during the acquisition time, an incubator gas chamber (Okolab) equipped with CO2 sensors, temperature unit and an active humidity controller was used. In order to label cell membranes, 2.5 μl of wheat germ agglutinin conjugated with Alexa Fluor 488 dye (λEx = 495 nm) was added to the cell samples and after 10 min of incubation at 37°C, 5% CO2, two washing steps were performed by using 500 μl of live-cell imaging solution (1×, Molecular Probes) at 37°C. ZnO NCs were used as labeled with Atto647 NHS ester dye, EVs were labeled in DiD and in DiO for sample treated with EVs and TNHs, respectively.
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