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**What is a T75 flask?**.
A T75 flask is a type of tissue culture flask commonly used in laboratory settings for cell culture. It is designed to provide a large surface area for cell attachment and growth, making it ideal for the propagation of mammalian cells.
**How to use a T75 flask:**.
1. **Prepare the flask**: Before use, sterilize the T75 flask by autoclaving or using a disinfectant such as ethanol. Be sure to work in a sterile environment, such as a laminar flow hood, to prevent contamination of the cell culture.
2. **Thaw the cells**: If using frozen cells, thaw them according to the protocol provided with the cells. Be gentle when handling the cells to maintain their viability.
3. **Seed the cells**: Carefully pipette the desired number of cells into the T75 flask, along with the appropriate growth medium. The number of cells seeded will depend on the specific cell line and experimental requirements.
4. **Incubate the flask**: Place the T75 flask in a cell culture incubator set to the appropriate conditions for the cells being cultured (e.g., temperature, CO2 levels). Allow the cells to attach to the flask surface and grow.
5. **Change the medium**: After the cells have had time to attach and grow, remove the old growth medium and replace it with fresh medium. This helps to provide the cells with essential nutrients and maintain their growth.
6. **Monitor the cells**: Regularly check the cells under a microscope to assess their growth and health. Look for signs of confluence (cells forming a monolayer) or any contamination that may be present.
7. **Passage the cells**: When the cells reach confluency in the T75 flask, they will need to be passaged into a new flask to prevent overgrowth. Follow the appropriate protocol for passaging the specific cell line being cultured.
8. **Dispose of the flask**: Once the cells have been harvested or the experiment is complete, dispose of the T75 flask properly according to biohazard waste disposal guidelines. Be sure to autoclave or disinfect the flask before discarding.
In conclusion, a T75 flask is a useful tool for cell culture in the laboratory, providing a large surface area for cell growth. By following these step-by-step instructions, researchers can effectively use T75 flasks to maintain and propagate mammalian cells for various experimental purposes.
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