Aseptic Techniques: Cell Culture Basics

Summary

This video script emphasizes the criticality of maintaining sterile environments in cell culture to prevent contamination. It outlines the necessity of personal protective equipment, proper hand hygiene, and cleaning protocols. The script details the correct use of a cell culture hood, including setup, handling of pipettes and media, and the importance of aseptic technique. It also highlights the proper disposal of contaminated items and the importance of cleanliness post-experimentation.

Takeaways

• 🔬 Maintaining a sterile environment is crucial for successful cell culture to prevent microorganism contamination.
• 🧼 Always wear personal protective equipment (PPE) when working in the lab, including closed-toe shoes and clothes covering your legs.
• 👀 Review the Material Safety Data Sheets (MSDS) before handling any media or reagents to ensure safety.
• 🧼 Wash hands thoroughly before handling cells or performing cell culture work to remove bacteria and dead skin particles.
• 🚫 Avoid using alcohol spray in areas with open flames due to fire hazards.
• 🧹 Clean the cell culture hood before and after use with 70% ethanol to maintain a sterile environment.
• 🛡️ Keep the cell culture hood work surface uncluttered to prevent accidental contamination and ensure proper airflow.
• 🔧 Use a pipette aid for proper handling of pipettes to avoid contamination during cell culture procedures.
• 🏺 Only open media, reagents, and supplies within the cell culture hood to maintain sterility.
• 🔄 Practice proper pipetting technique and avoid touching non-sterile surfaces to prevent cross-contamination.
• 🗑️ Dispose of any contaminated pipettes immediately and do not reuse them to maintain a sterile field.

Q & A

Why is it crucial to keep cells free from microorganism contamination during cell culture?

Keeping cells free from microorganism contamination is crucial because contamination can disrupt the cell culture's integrity, leading to inaccurate experimental results or complete loss of the culture.

What is the role of personal protective equipment (PPE) in cell culture?

PPE is essential for protecting both the researcher and the cell culture from contamination. It includes items like lab coats, gloves, and safety goggles, which prevent the introduction of contaminants from the researcher to the cell culture.

Why is it important to review the Material Safety Data Sheet (MSDS) before working with media or reagents?

Reviewing the MSDS is important to understand the potential hazards of the chemicals being used, how to handle them safely, and what precautions to take to minimize risk to both the researcher and the cell culture.

What does it mean to work in a sterile technique?

Working in a sterile technique means performing procedures in a way that minimizes the risk of introducing contaminants into the cell culture. This includes working slowly and deliberately, using sterile equipment, and maintaining a clean work area.

Why should you clean the cell culture hood before and after use?

Cleaning the cell culture hood before and after use helps to prevent the buildup of contaminants that could otherwise be introduced into the culture during the next session.

What is the significance of using 70% ethanol for hand washing in a cell culture lab?

A 70% ethanol wash is effective at killing microorganisms on the skin. It is used to remove bacteria and other contaminants before handling cells or culture materials to prevent their introduction into the culture.

How should you handle items placed in the cell culture hood to maintain sterility?

Items placed in the cell culture hood should be cleaned with ethanol before placement. They should be handled with sterile techniques, avoiding contact with non-sterile surfaces, and placed in the hood with a forward motion to not disrupt the airflow.

Why is it advised not to overcrowd the work surface in a cell culture hood?

Overcrowding the work surface can increase the risk of contamination through accidental touching and disrupt the airflow, which is necessary to maintain a sterile field within the hood.

What is the correct way to open a pipette package without contaminating the pipette?

The correct way is to grasp the pipette high on the neck, insert it into the pipette aid, turn the desired measurement marks toward you, and then discard the wrapper without letting the pipette touch anything non-sterile.

Why is it important to avoid touching the inside edge of the cap when handling media or reagents?

Touching the inside edge of the cap can contaminate it, as this part comes into direct contact with the media or reagents. It's crucial to replace the cap as soon as possible to maintain sterility.

How should you transfer large volumes of liquid between containers in a sterile manner?

The best way to transfer large volumes is to use a sterile pipette. If that's not possible, pour quickly and deliberately using the front corner of the bottle to channel the liquid, ensuring accuracy and minimizing the risk of contamination.