Sterile Technique
Summary
TLDRThis instructional video from Addgene emphasizes the importance of aseptic technique in lab sciences. It covers personal protective equipment, setting up a clean workspace, and maintaining sterility. Key points include wearing lab coats and gloves, keeping workspaces organized, and using alcohol and flame to sterilize tools. The video also demonstrates proper handling of agar dishes and bottles to prevent contamination, ensuring a safer and more effective laboratory practice.
Takeaways
- 🔬 Aseptic technique is crucial for success in fields like microbiology, molecular biology, and cell biology.
- 🧼 Personal protective equipment (PPE) such as lab coats, gloves, and safety glasses are essential for maintaining aseptic conditions.
- 👕 Ensure arms and wrists are covered to minimize the risk of contaminating samples with body flora.
- 🧤 Gloves protect both the user and the material from contamination; change them when necessary.
- 🧹 Clear the workspace of unnecessary materials and keep clean and dirty items separate to maintain a clean working environment.
- 🔥 Use a Bunsen burner or candle jar to create a sterile field, and keep the surrounding area free of flammable materials.
- 🍶 Wipe down workspace and equipment with 70% alcohol to disinfect surfaces before starting work.
- 🔍 Inspect all materials for signs of contamination or compromised sterility, including checking autoclaved items and sealed containers.
- 🔥 Sterilize metal objects like forceps by flaming after dipping in 70% alcohol for reuse in the lab.
- 🧫 When working with agar dishes, hold the lid at an angle to inoculate to prevent contamination from body flora.
- 🌐 Visit Addgene's website for more resources on aseptic technique, protocols, and scientific sharing.
Q & A
What is aseptic technique and why is it important in scientific fields?
-Aseptic technique is a critical skill in fields such as microbiology, molecular biology, and cell biology, which involves maintaining a sterile environment to prevent contamination of samples and reagents.
What personal protective equipment is recommended for working in a lab?
-Appropriate personal protective equipment includes a lab coat, gloves, and safety glasses if necessary.
How can body flora be a source of contamination in a lab setting?
-Body flora can contaminate reagents and samples, so it's important to cover arms and wrists with a cuffed lab coat or disposable arm cuffs to reduce this risk.
What are some best practices for maintaining glove cleanliness in the lab?
-To keep gloves clean, avoid touching skin, hair, cell phones, writing utensils, or other unsterilized objects, and always change gloves if they come into contact with something unsterile or when starting a new task.
How should a workspace be organized to minimize contamination risks?
-The workspace should be cleared of unnecessary materials, with clean and dirty items separated and within arm’s reach. It's recommended to work from left to right, with clean items on the left and waste bins on the right.
Why is it important to wipe down workspaces and pipettes with 70% alcohol?
-Wiping down workspaces and pipettes with 70% alcohol helps to disinfect surfaces and prevent contamination of materials and samples.
How can you check if autoclaved items have maintained their sterility?
-Check autoclaved items for color changes in the indicator tape to ensure sterility. For foil-covered items, check for tears or holes, and for capped items, ensure the cap is tightly sealed.
What is the proper way to handle foil when working with sterile containers?
-Gently loosen the edges of the foil before starting work, lift it with one hand to keep it upright, and hold it in your hand rather than putting it down to prevent contamination of the sterile side.
How should you handle glass bottles containing sterile liquids to maintain sterility?
-Gently loosen the cap before starting, remove it with one hand without placing it on the lab bench, and pass the bottle neck through a flame before and after use to sterilize it.
What method can be used to sterilize metal objects like forceps and spreaders in the lab?
-Dip the metal objects in 70% alcohol, pass the alcohol-dipped portion through a flame to ignite and burn off the alcohol, and cool the hot tip on a sterile surface before use if it's for live cultures.
How can you reduce the risk of body flora contamination when working with agar plates?
-Hold the lid of the agar dish at a 75-90 degree angle towards you and inoculate from behind to form an additional barrier against contamination.
Outlines
🔬 Aseptic Technique Essentials
Meghan, a scientist at Addgene, introduces aseptic technique as a vital skill for fields like microbiology, molecular biology, and cell biology. The video covers key strategies for aseptic technique, emphasizing the importance of personal protective equipment (PPE) such as lab coats, gloves, and safety glasses. It advises on covering arms and wrists to minimize contamination risk, changing gloves when necessary, and setting up a clean workspace. The video also instructs on maintaining sterility, checking materials for contamination, and proper handling of agar dishes, glass bottles, and metal instruments. It concludes by promoting Addgene's resources for further learning.
Mindmap
Keywords
💡Aseptic technique
💡Personal protective equipment (PPE)
💡Sterility
💡Contamination
💡Lab coat
💡Gloves
💡Workspace
💡Bunsen burner
💡Alcohol
💡Indicator tape
💡Agar dishes
Highlights
Aseptic technique is crucial for success in microbiology, molecular biology, and cell biology.
Personal protective equipment includes lab coats, gloves, and safety glasses.
Body flora can cause contamination in reagents; cover arms and wrists to reduce risk.
Gloves protect both the user and the material from contamination.
Change gloves if touching unsterile objects or starting a new task.
Clear workspace of unnecessary materials and set up materials within arm’s reach.
Use a bunsen burner or candle jar with caution, keeping the surrounding area clear.
Wipe down workspace and pipettes with 70% alcohol before starting work.
Inspect materials for contamination and ensure sterility, checking autoclaved items' indicator tape.
Handle foil-covered bottles and beakers carefully to maintain sterility.
Work quickly to minimize exposure of sterile items to the environment.
Sterilize metal objects like forceps and spreaders with alcohol and a flame.
Cool hot instruments before contacting live cultures to avoid killing the specimen.
When working with agar dishes, do not fully remove the lid to reduce body flora contamination.
Inoculate agar plates from behind, using the lid as a barrier against contamination.
For more information on aseptic technique, visit Addgene’s website for videos, protocols, and blog posts.
Transcripts
Hi, I'm Meghan, a scientist at Addgene
Aseptic technique is a critical skill for success in numerous fields such as microbiology,
molecular biology or cell biology.
In this instructional video we will provide an overview of some of the key strategies
of aseptic technique, including personal protective equipment, setting up a clean workspace and
maintaining sterility while working.
Before beginning any experiment make sure that you are wearing the appropriate personal
protective equipment, including a lab coat, gloves and, if necessary, safety glasses.
As we mentioned during our protocol on “Getting Started With Tissue Culture”, body flora
can be a common cause of contamination in reagents.
To reduce the risk of contaminating your samples with body flora, ensure that your arms and
wrists are covered by a cuffed lab coat or disposable arm cuffs.
While gloves are meant to protect users from the material they are working with, they also
protect the material from user contamination.
To keep your gloves clean, avoid touching your skin and hair, as well as your cell phone,
writing utensils, or other unsterilized objects.
Always change your gloves if you touch something unsterile or are starting a new task.
Before beginning any task, clear the workspace of unnecessary materials.
Set up all materials so that they are in arm’s reach with clean and dirty items clearly separated.
We recommend working from left to right with all clean items stored on the left and waste
bins to the right.
If you are going to be using a bunsen burner or candle jar, make sure that the space around
the burners are free of loose papers or laboratory wipes.
Finally, wipe down the workspace and pipettes with 70% alcohol and allow it to evaporate.
Before beginning a task, examine all of your materials for signs of contamination or signs
that the sterility may have been compromised.
Check all autoclaved items to ensure that the indicator tape has changed colors.
For items that are covered with foil check for tears or holes for capped items, ensure
that the cap has been tightly sealed.
When working with agar dishes, examine the dishes for signs of microbial growth.
Foil is often used as a cover for bottles and beakers.
Before beginning your work, gently loosen the edges of the foil, so it is easier to
remove.
When ready, gently lift the foil with one hand and keep it upright, to prevent contamination
of the sterile side.
While working, keep the foil in your hand rather than putting it down on the lab bench.
Work quickly so that the beaker does not sit uncovered for an extended period of time.
When working in the lab with sterile liquids in glass bottles, gently loosen the cap before
starting.
Using one hand, remove the cap from the bottle.
To prevent the sterile side of the cap from becoming contaminated, do not put the cap
on the lab bench.
Instead hold it in your hand upright as you work.
While holding the bottle at an angle, gently pass the neck of the bottle through the flame.
Before recapping, pass the neck of the bottle through the flame a second time.
If you are going to use the bottle again, you can leave the cap loosely sitting on the
top of the bottle.
Once you are finished, however, be sure to cap the bottle tightly.
Metal objects such as forceps and spreaders are used and reused frequently in the lab.
While these items can be sterilized in an autoclave, it is often more convenient to
disinfect with alcohol and a burner.
To sterilize, dip the portion of the instrument that will be used for the procedure in 70%
alcohol.
Pass the alcohol-dipped portion of the instrument through the flame of the burner to ignite
and then allow the alcohol to burn off.
If you are going to use the instrument on a live culture, such as bacteria, touch the hot tip to the inside
of a sterile tube or the surface of an uninoculated culture to cool before contacting the specimen.
Agar dishes are used in the lab to culture bacteria such as E. coli.
To reduce the risk of body flora contamination when working with agar plates, do not fully
remove the lid of the agar dish.
Instead, hold the lid at 75-90 degree angle toward you and inoculate from behind.
The lid will form an additional barrier against unwanted contamination.
Thank you for watching this aseptic technique video.
For more useful videos, protocols or blog posts please visit Addgene’s website.
Addgene - a better way to share science.
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