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1.5: Aseptic Technique Procedure

  • Page ID
    130342
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      The Importance of Aseptic Technique, Procedure 

     

    Name: _______________________________________

    Course Section: _______________________________

     

    The following activity will help demonstrate the importance of aseptic technique, sterilization, and the ubiquitous nature of microbes. You will collect samples from your skin and environment, inoculate the microbes on nutrient agar, and observe patterns of microbial growth. Read the following procedure carefully before starting the activity.

     

    Materials

    • 2 Nutrient Agar Plates
    • Soap and Water
    • 60% Alcohol Hand Sanitizer
    • 1 Sterile Swab
    • Parafilm  
    • Sharpies
    • DI Water

     

    clipboard_e9379ff77f86c0bdc2e05508715e241cc.png clipboard_e5b62bbc265513403a08f24ea3287369f.png
    How To Label Nutrient Agar Plate #1 How To Label Nutrient Agar Plate #2

     

     

    Procedure: Nutrient Agar Plate #1

    Step 1: Get into groups of 2. 

    Step 2: On the bottom of Agar Plate #1, use a sharpie to split the plate into 4 quadrants, creating a "crosshair" shape in the middle of the plate. Label each quadrant 1 through 4.

    • Decide who will wash their hands with soap and water and who will sanitize their hands. Record this in the results table. 
    • Note, quadrants 1 and 2 will correspond to soap and water usage. Quadrants 3 and 4 will correspond to hand sanitizer usage. 
    • Do not wash, clean, or sanitize your hands during this step. 

    Step 3: One group member will press the pad of their index finger in the quadrant marked "1". The other group member will press the pad of their index finger in the quadrant marked "3".

    • Do not wash, clean, or sanitize your hands during this step. 
    • Be gentle with the agar, pressing too hard will break it. 

    Step 4: Wash your hands with soap and water or use hand sanitizer.  

    Step 5: With clean hands, use the same index finger and gently press it into quadrant "2" if you used soap and water or quadrant "4" if you used hand-sanitizer. 

    Step 6: Wrap the plate in parafilm and place it in the 37°C incubator. Plates should be inverted: agar-side up and lid-side down. 

     

     

    Procedure: Nutrient Agar Plate #2

    Step 1: You and your partner will select a random surface to swab. This item is called the "Wild Card" in the results table.

    Step 2: Once you have picked a surface, make sure you write down the name of the item on Agar Plate #2 and in the results table.

    Step 3: Add a few drops of sterile DI water to your swab, this will help pick up microbes from your surface of interest.

    Step 4: Swipe the swab over your selected surface/item multiple times.

    Step 5: Drag your swab across the surface of the agar plate in a zig-zag (back and forth) pattern across the entire plate surface. 

    • Make sure you keep the lid slightly over the agar plate when applying the swab. This will help decrease potential contamination. 

    Step 6:  Wrap the plate in parafilm and place it in the 37°C incubator. Plates should be inverted: agar-side up and lid-side down. 

     

     

     

     

     

    Results

    In table below, describe the microbial growth present for each quadrant and the Wild Card sample. Discuss the color, quantity, shape, and other characteristics present for the colonies that grew.   

    Results Table
      Observations
      Pre-Wash  Post-Wash

    Soap and Water

    Name:

    Quadrant 1:

     

     

     

     

     

     

    Quadrant 2:

     

     

     

     

    60% Alcohol Hand Sanitizer 

    Name: 

    Quadrant 3:

     

     

     

     

     

     

    Quadrant 4:

     

     

     

     

    Wild Card

    Item Sampled: 

     

     

     

     

     

     

     

     

     

    General Questions

    1. Which sanitization method was more effective, using soap and water or hand sanitizer? Explain your reasoning. 

     

     

     

     

    2. Why is aseptic technique important in microbiology labs?

     

     

     

     

    3. What is the purpose of using an incinerator? 

     

     

     

    4. Why do we invert petri dishes in the incubator?

     

     

     

     

    5. List 3 potential sources of contamination that may occur in microbiology labs. 

    • w
    • w
    • w

     

     

    6. When aseptic technique is applied to a healthcare setting it is called

     

         __________________________     _________________________.

     

    7. Why is it important to keep a patient's skin healthy during their hospitalization?

     

     

     

     

     

    8. Why does Scabies target skin folds and not other areas of the body?

     

     

     

     

     

    9. How can healthcare workers prevent Scabies from developing in their patient?

     

     

     

     

    Attributions

    "Microbiology Laboratory Manual: Labs, 1.7 Aseptic Technique" by Dr. Rosanna HartlineLibreTexts: Biology, West Hills College Lemoore is licensed under CC BY-NC 4.0

    "Environmental Infection Control Guideline: Part F, Environmental Sampling " by Centers for Disease Control and PreventionUse of CDC Materials Statement is in the Public Domain

    Image Citations 

    The images presented in the "Procedure" were Modified From:


    1.5: Aseptic Technique Procedure is shared under a CC BY 4.0 license and was authored, remixed, and/or curated by LibreTexts.

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