Some Aseptic Techniques You Can Use In Your Tissue Culture Process

Summary

This video demonstrates essential asepticTranscript summary generation techniques for plant tissue culture, focusing on maintaining a sterile work environment. It covers cleaning the workspace with 70% isopropyl alcohol or ethanol, using a glass bead sterilizer as an alternative to a Bunsen burner, and proper handling of forceps while splitting plants. The tutorial specifically works with Sterogen ribbons, popular in the aquarium hobby, and emphasizes safety measures like wearing masks and gloves. Viewers learn how to propagate multiple plant cultures efficiently while minimizing contamination, with tips on personal preferences in technique and maintaining cleanliness for future work. Practical and accessible for hobbyists and beginners alike.

Takeaways

• 😀 Clean your work area with 70% isopropyl alcohol or 70% ethanol before starting tissue culture.
• 😀 If you don't have access to a laminar flow hood, using an air filter works as a good alternative.
• 😀 Always pre-heat your laminar flow hood 30 minutes before use to ensure proper sterile conditions.
• 😀 Use a glass bead sterilizer as an alternative to a bunsen burner for sterilizing forceps and tools.
• 😀 When sterilizing forceps with a glass bead sterilizer, maintain the temperature at 250°C.
• 😀 Sterogen ribbons are commonly used in aquarium plant hobbies and can be cultured using these techniques.
• 😀 Always wear a mask to avoid contamination when working with tissue culture.
• 😀 Use 70% isopropyl alcohol or ethanol to clean your hands and tools during the process.
• 😀 Gloves are optional for using the glass bead sterilizer but not recommended when using a bunsen burner.
• 😀 After sterilizing the forceps, let them cool naturally or use media to speed up the cooling process.
• 😀 To propagate cultures, split plants with sterile tools, and always sterilize tools between uses.
• 😀 Avoid worrying about others' techniques if what you're doing works effectively for you.

Q & A

Why is it important to clean the working area with 70% isopropyl alcohol or ethanol in tissue culture?

Cleaning the working area with 70% isopropyl alcohol or ethanol helps to eliminate contaminants such as bacteria and fungi, ensuring a sterile environment for tissue culture work. This prevents any unwanted microbial growth that could affect the plants being cultured.

What should you do if you don't have access to a laminar flow hood?

If you don’t have access to a laminar flow hood, you can use an air filter as an alternative. Although it’s not as effective as a laminar flow hood, it can still provide some level of air filtration to reduce contamination risks.

How long before using a laminar flow hood should you turn it on?

It’s recommended to turn on the laminar flow hood at least 30 minutes ahead of time to allow it to properly filter and purify the air before you begin your work.

Why is a glass bead sterilizer used instead of a Bunsen burner?

A glass bead sterilizer is used in this video because it’s a safer alternative, especially when the video is being filmed. It allows the user to sterilize forceps without the risk of fire hazards associated with a Bunsen burner.

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The glass bead sterilizer should be set to a temperature above 250°C to effectively sterilize tools like forceps.

What is the purpose of wearing a mask and using alcohol to clean hands in tissue culture?

Wearing a mask helps reduce the risk of introducing contaminants from breathing or talking, while cleaning your hands with 70% isopropyl alcohol or ethanol ensures that any microorganisms on your hands are eliminated before handling sterile tools or cultures.

Can gloves be worn while using a glass bead sterilizer?

While gloves can be worn when using a glass bead sterilizer, they are not recommended if using a Bunsen burner, as the heat from the flame may cause discomfort or injury.

What should you do to cool down sterilized forceps if you don’t have access to a laminar flow hood?

If you don’t have access to a laminar flow hood, you can use the media to cool down the sterilized forceps more quickly. The media acts as a cooling agent, allowing you to handle the sterilized tools sooner.

How many plants do you need to make a new culture?

You only need one plant to create a new culture. By splitting the plant into smaller pieces, you can propagate multiple cultures from a single plant.

What is the importance of repeatedly sterilizing the forceps during the process?

It is crucial to sterilize the forceps between each use to avoid cross-contamination. By placing the forceps back into the glass bead sterilizer for 10 to 15 seconds, you ensure that no unwanted microorganisms are transferred to the plants during the process.