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T75 flask cell culture is a common technique in laboratory settings for growing and maintaining cells for research purposes. Here are 10 important questions you should know about T75 flask cell culture.
### What is a T75 flask?
A T75 flask is a type of cell culture flask that has a total volume of 75 ml. It is commonly used for cell culture experiments in research laboratories.
### How do you prepare a T75 flask for cell culture?
1. Start by sterilizing the flask using an autoclave or by using a sterile filter.
2. Add the appropriate cell culture medium to the flask, making sure it covers the entire surface area.
3. Place the flask in a 37°C incubator to warm the medium.
### How do you seed cells in a T75 flask?
1. Remove the T75 flask from the incubator and aspirate the medium.
2. Add the desired number of cells in suspension to the flask.
3. Gently swirl the flask to evenly distribute the cells.
### How often should the medium be changed in a T75 flask?
The medium in a T75 flask should be changed every 2-3 days to provide the cells with fresh nutrients and remove waste products.
### How do you split cells in a T75 flask?
1. Aspirate the medium from the flask and wash the cells with phosphate-buffered saline (PBS).
2. Add trypsin-EDTA to detach the cells from the flask.
3. Transfer the cell suspension to a new T75 flask with fresh medium.
### How do you store a T75 flask?
T75 flasks should be stored in a 5% CO2 incubator at 37°C to maintain the optimal conditions for cell growth.
### What are some common troubleshooting tips for T75 flask cell culture?
1. Check for contamination regularly by inspecting the cells under a microscope.
2. Ensure proper handling and aseptic techniques to prevent contamination.
3. Monitor cell morphology and growth rate to ensure healthy cell culture.
### How do you passaged cells in a T75 flask?
Passaging cells in a T75 flask involves detaching the cells from the flask, counting them, and seeding them into new flasks at a lower density to allow for continued growth.
### Conclusion.
Understanding the basics of T75 flask cell culture is essential for successful cell culture experiments in the laboratory. By following proper techniques and protocols, researchers can maintain healthy cell cultures for their experiments.
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