Summary
Highlights
Successful cell culture relies on quality cells and reagents, sterile technique, experience, and the right equipment. Proper setup is crucial, with all necessary items close at hand.
Key equipment includes a cell culture hood for a sterile environment, an incubator, water bath, centrifuge, refrigerator/freezer, hemocytometer, microscope, cell culture flasks, medium, reagents, pipettes, and waste containers. A cell counting machine or hemocytometer is used for cell counting.
Incubators provide optimal conditions for cell growth, typically 37°C with 5-7% CO2 and high humidity for mammalian cells. Flasks should be evenly spaced to ensure proper air circulation and gas exchange. Media and reagents must be stored according to label instructions, protected from light and extreme temperatures.
Reagents can be warmed to 37°C or room temperature. When using a water bath, keep bottles upright and use bottle weights or metallic bath beads as a cleaner alternative. Avoid prolonged warming, as it can degrade essential components.
Maintain proper ergonomic posture and avoid breathing or talking into the hood. Always wipe the hood and all items within it with 70% ethanol before and after use. Arrange items strategically for efficient and sterile workflow, placing cell plates in the center.
Transport cells carefully to prevent damage to the monolayer and splashing, which can lead to contamination. For flasks with unvented caps, tighten them before removal from the incubator. Regularly view cultures under a microscope to check cell health, growth, and contamination. Verify cell identity to ensure you are working with the correct culture.