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    MP Lab 1 2

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    This document provides instructions for performing bacterial smears and gram staining. It describes the necessary materials, which include glass slides, saline solution, inoculating needles, and stains. The procedures involve placing a bacterial sample onto a slide, fixing it, and then staining it with crystal violet, iodine, and safranin to determine if the bacteria is gram positive or gram negative based on its color. Safety precautions like personal protective equipment and proper disposal of materials are also outlined.

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    MP Lab 1 2

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    This document provides instructions for performing bacterial smears and gram staining. It describes the necessary materials, which include glass slides, saline solution, inoculating needles, and stains. The procedures involve placing a bacterial sample onto a slide, fixing it, and then staining it with crystal violet, iodine, and safranin to determine if the bacteria is gram positive or gram negative based on its color. Safety precautions like personal protective equipment and proper disposal of materials are also outlined.

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    MP-LAB-1-2

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    This document provides instructions for performing bacterial smears and gram staining. It describes the necessary materials, which include glass slides, saline solution, inoculating needles, and stains. The procedures involve placing a bacterial sample onto a slide, fixing it, and then staining it with crystal violet, iodine, and safranin to determine if the bacteria is gram positive or gram negative based on its color. Safety precautions like personal protective equipment and proper disposal of materials are also outlined.

    Copyright:

    © All Rights Reserved
    Download as DOCX, PDF, TXT or read online from Scribd
    Download as docx, pdf, or txt
    18 views4 pages

    MP Lab 1 2

    Uploaded by

    Fame Who
    This document provides instructions for performing bacterial smears and gram staining. It describes the necessary materials, which include glass slides, saline solution, inoculating needles, and stains. The procedures involve placing a bacterial sample onto a slide, fixing it, and then staining it with crystal violet, iodine, and safranin to determine if the bacteria is gram positive or gram negative based on its color. Safety precautions like personal protective equipment and proper disposal of materials are also outlined.

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    © All Rights Reserved
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    of 4

    ADVENTIST MEDICAL CENTER COLLEGE

    Department of Medical Technology


    BACTERIOLOGY LABORATORY ACTIVITY No. 1

    BACTERIAL SMEAR

    Materials: The materials for bacterial smear procedure are the following:
    1 2 3 Checklist:
    1. Sterile glass slide
    2. Sterile Inoculating needle
    3. Bunsen burner/alcohol lamp
    4. Match
    5. Sterile Natural Saline Solution
    6. Sterile applicator stick
    7. Bacterial colony
    8. Specimen

    A. Bacterial smear from a colony:


    1 2 3 Checklist
    1. Put on your personal protective equipment such as mask, gloves, and lab gown.
    2. Prepare the materials and make sure they are within arms’ reach.
    3. Open the cover of the sterile glass slide and label using a sticker and pencil.
    4. Place a small drop of sterile NSS on a sterile glass slide.
    5. Open the agar with the bacterial colony growth, and with the use of sterile inoculating
    needle/applicator stick, fish out a portion of the colony to be stained.
    6. and gently Emulsify onto the drop of NSS on the sterile glass slide.
    7. Air dry.
    8. Heat fix by gently passing the smear on a flame three times.
    9. Place the smear on the staining rack.
    10. Perform after care, by discarding into the infectious waste container and sterilizing the materials and
    biosafety cabinet.

    B. From a specimen:
    1 2 3 Checklist:
    1. Put on personal protective equipment such as mask, gloves, and lab gown.
    2. Prepare the materials and make sure they are within arms’ reach.
    3. Open the cover of the clean sterile glass slide and label using a sticker and pencil.
    4. Place a small drop of sterile NSS on a sterile glass slide.
    5. With the use of sterile inoculating needle/applicator stick/sterile swab, get a small portion of the
    specimen.
    6. Gently apply the area of the sterile inoculating needle/applicator stick/sterile swab on the glass slide
    but do not rub it hard so that the cells wont get distorted.
    7. Air dry.
    8. Heat fix by gently passing the smear on a flame three times.
    9. Place the smear on the staining rack.
    10. Perform after care, by discarding into the infectious waste container and sterilizing the materials and
    biosafety cabinet.
    ADVENTIST MEDICAL CENTER COLLEGE
    Department of Medical Technology
    BACTERIOLOGY LABORATORY ACTIVITY No. 2

    GRAM STAINING

    Most bacteria are classified as Gram-positive or Gram-negative according to their response to the Gram staining
    procedure. The Gram stain depends on the ability of certain bacteria (Gram-positive) to retain the crystal violet-iodine
    complex after decolorization with acetone alcohol (purple). Gram-negative bacteria do not retain the dye-iodine complex
    and become transluscent (colorless) after decolorization but takes up the red counterstain safranin. Thus, Gram-positive
    bacteria look purple under the microscope and Gram-negative bacteria look red.

    The Materials for gram staining procedure are the following:


    1 2 3 Checklist:
    1. Air-dried Bacterial smear
    2. Gram staining set,
    3. Microscope
    4. Staining Rack
    5. Faucet with running water
    6. Caliper

    The procedure for the gram staining.


    1 2 3 Checklist:
    1. Put on personal protective equipment such as mask, gloves, and lab gown.
    2. Flood the smear with crystal violet for one minute. Wash with water.
    3. Flood the smear with Gram’s iodine for one minute. Wash with water.
    4. Decolorize with acetone-alcohol solution (usually 1-3 seconds). Wash with water.
    5. Counterstain with safranin for 30 seconds to 1 minute. Wash with water.
    6. Using a caliper place the gram-stained smear on the staining rack and air dry.
    7. Examine under oil immersion objective and evaluate the bacteria’s Gram reaction.
    8. Check to see whether Gram-positive or Gram-negative. Gram positive is color purple, while gram
    negative is color pink.
    9. Check for the bacterial morphology whether cocci or bacilli. Cocci are sphere in shape, while bacilli are
    rod shaped.
    10. And check for the bacterial arrangement such as in chains, pairs, or clusters.
    11. Perform after care, by discarding into the infectious waste container and sterilizing the materials and
    biosafety cabinet.

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