Podcast
Questions and Answers
What is the primary aim of using aseptic techniques in a microbiology laboratory?
What is the primary aim of using aseptic techniques in a microbiology laboratory?
- To alter the genetic makeup of the microorganisms being studied.
- To accelerate the growth rate of microorganisms.
- To minimize contamination of cultures and the lab environment. (correct)
- To introduce new microorganisms into the culture.
Which of the following is the MOST critical reason for employing aseptic techniques when culturing bacterial cells?
Which of the following is the MOST critical reason for employing aseptic techniques when culturing bacterial cells?
- To reduce the visibility of bacterial colonies under a microscope.
- To promote the adhesion of bacterial cells to lab equipment.
- To ensure the bacterial cells mutate rapidly.
- To prevent contamination of the specific microorganism and the surrounding environment. (correct)
Which of the following practices is MOST important for preventing contamination when working with bacterial cultures?
Which of the following practices is MOST important for preventing contamination when working with bacterial cultures?
- Using a brightly lit workspace.
- Recycling used culture plates immediately.
- Playing music during experiments.
- Wearing personal protective equipment (PPE). (correct)
When preparing to work with aseptic techniques, what is the purpose of sterilizing the lab bench with 70% ethanol?
When preparing to work with aseptic techniques, what is the purpose of sterilizing the lab bench with 70% ethanol?
Which action BEST minimizes contamination when opening sterile bottles or containers during an experiment?
Which action BEST minimizes contamination when opening sterile bottles or containers during an experiment?
Why is it crucial to avoid passing hands or arms over open bottles or plates during aseptic procedures?
Why is it crucial to avoid passing hands or arms over open bottles or plates during aseptic procedures?
What is the primary reason for performing experiments as quickly as possible when using aseptic techniques?
What is the primary reason for performing experiments as quickly as possible when using aseptic techniques?
In the spread plate method, what characteristic of the bacteria is most important for easy counting and isolation?
In the spread plate method, what characteristic of the bacteria is most important for easy counting and isolation?
What is the purpose of using a sterilized spreader with a smooth surface in the spread plate technique?
What is the purpose of using a sterilized spreader with a smooth surface in the spread plate technique?
Why are plates required to be dry and at room temperature for the spread plate technique?
Why are plates required to be dry and at room temperature for the spread plate technique?
Which of the following is a crucial step when removing a sterile spreader from its packaging to maintain sterility?
Which of the following is a crucial step when removing a sterile spreader from its packaging to maintain sterility?
During the spread plate method, what determines appropriate selection of bacterial colonies?
During the spread plate method, what determines appropriate selection of bacterial colonies?
When picking a colony using a sterile loop, what step ensures the loop remains sterile?
When picking a colony using a sterile loop, what step ensures the loop remains sterile?
When inoculating a bacterial colony into a tube, what is the correct procedure after inserting the loop into the tube?
When inoculating a bacterial colony into a tube, what is the correct procedure after inserting the loop into the tube?
After inoculating a tube, why should the cap NOT be tightly sealed?
After inoculating a tube, why should the cap NOT be tightly sealed?
For solid waste contaminated with bacteria, such as bacterial spreaders and plates, what is the standard disposal method?
For solid waste contaminated with bacteria, such as bacterial spreaders and plates, what is the standard disposal method?
What is the correct method for disposing of liquid bacterial cultures and unused liquid media (LB)?
What is the correct method for disposing of liquid bacterial cultures and unused liquid media (LB)?
What does the term 'aseptic technique' primarily refer to in a microbiology setting?
What does the term 'aseptic technique' primarily refer to in a microbiology setting?
Why is aseptic technique considered a compulsory skill in microbiology research?
Why is aseptic technique considered a compulsory skill in microbiology research?
Which external element is LEAST likely to be a direct source of contamination in a cell culture?
Which external element is LEAST likely to be a direct source of contamination in a cell culture?
A researcher consistently finds unexpected contaminants in their cultures despite diligently following usual aseptic protocols. Considering potential ‘Sources of Contamination’, what should they FIRST examine?
A researcher consistently finds unexpected contaminants in their cultures despite diligently following usual aseptic protocols. Considering potential ‘Sources of Contamination’, what should they FIRST examine?
Select the statement that is an INCORRECT depiction of aseptic technique principles:
Select the statement that is an INCORRECT depiction of aseptic technique principles:
A researcher is preparing to perform a bacterial inoculation. They have their PPE on, the workspace is clean, and all materials are ready. What is the MOST IMMEDIATE next step?
A researcher is preparing to perform a bacterial inoculation. They have their PPE on, the workspace is clean, and all materials are ready. What is the MOST IMMEDIATE next step?
A lab technician finds fungal contamination in a supposedly sterile batch of LB media after autoclaving. What might be the MOST PROBABLE cause, assuming the autoclave itself is functioning correctly?
A lab technician finds fungal contamination in a supposedly sterile batch of LB media after autoclaving. What might be the MOST PROBABLE cause, assuming the autoclave itself is functioning correctly?
A researcher is using the spread plate method to quantify bacterial colonies. After plating a diluted sample, they count 25 colonies on a plate with a $1:10,000$ dilution. Given the formula: $\text{Number of colonies} \times \text{reciprocal of dilution} = \text{bacteria/ml}$, calculate the number of bacteria/ml in the original sample.
A researcher is using the spread plate method to quantify bacterial colonies. After plating a diluted sample, they count 25 colonies on a plate with a $1:10,000$ dilution. Given the formula: $\text{Number of colonies} \times \text{reciprocal of dilution} = \text{bacteria/ml}$, calculate the number of bacteria/ml in the original sample.
Flashcards
Aseptic Technique
Aseptic Technique
A method that involves target-specific practices and procedures under suitably controlled conditions to reduce contamination from microbes.
Why use aseptic technique?
Why use aseptic technique?
To prevent contamination of the specific microorganism being worked with and to prevent contamination of the room and personnel.
Spread plate method
Spread plate method
A technique to plate a liquid sample containing bacteria so that the bacteria are easy to count and isolate.
PPE Required
PPE Required
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Prepare the Lab Bench
Prepare the Lab Bench
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Study Notes
- Aseptic technique is a method involving target-specific practices and procedures under controlled conditions to reduce contamination from microbes
- It is essential for microbiology research and prevents contamination of microorganisms being cultured and contamination of the environment
Why Use Aseptic Technique
- Prevents specific microorganism contamination
- Prevents contamination of the room and personnel
Contamination Sources
- Personnel
- Facilities
- Equipment
- Utilities
- Materials
- Processes
- Sources can include flow of personnel, materials, waste, HVAC, assembly, cleaning, and sterilization
- Potential contaminants: water, gasses, raw materials, reusable resin, and membrane filters
Guidelines for Performing Aseptic Technique
- Close doors and windows
- Wash hands
- Wear PPE (lab coat, gloves, safety glasses)
- Pull long hair back
- Sterilize the lab bench, outside of bottles, and gloves with 70% ethanol
- Arrange the lab bench for easy reach
- Use sterile containers, pipettes, pipette tips, and solutions
- Only open bottles when needed and recap immediately
- Open bacterial plates away from you
- Avoid passing hands or arms over bottles or plates
- Work with a partner
- Refrain from talking, singing, or whistling during sterile procedures
- Perform experiments quickly to minimize contamination
Spread Plate Method
- Involves using a sterilized spreader with a smooth surface made of metal or glass to apply a small amount of bacteria suspended in a solution over a plate
- The plate needs to be dry and at room temperature so that the agar can absorb the bacteria more readily
Steps for Testing the Sterility of LB Media Using Aseptic Technique
- Put on personal protective equipment, including a lab coat, gloves, close-toes shoes and glasses
- Clear and sanitize the lab bench, and arrange reagents and supplies
- Remove media from the container
- Dispense media onto the LB Plate and open the plate using the 'clamshell' method on one-side
- Remove a sterile spreader at 'handle' end, then spread the media
Bacterial Inoculation Steps
- Pick an RFP+ or GFP+ bacterial colony and remove a sterile inoculating loop at 'handle' end being careful to not touch the loop
- Open the plate using the ‘clamshell' method, pick a colony using the sterile loop, and then open a tube that contains the LB
- Set the lid on the sanitized bench top inserting the loop into the tube and swirl gently, and remove the loop before capping the tube to the loose setting
Proper Bacterial Waste Disposal
- Autoclave biohazard bags for solid waste, like bacterial spreaders, inoculating loops, and bacterial plates
- Use a 10% Bleach solution for 10 minutes for liquid waste, like liquid bacterial cultures and unused liquid LB media, then pour down sink and flush with water
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Description
Understand the importance of aseptic techniques in microbiology for preventing contamination of cultures and the environment. Learn about potential contamination sources like personnel, equipment, and materials. Follow guidelines such as hand washing, PPE usage, and sterilization to maintain a sterile work environment.
