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    W1BACT211LAB

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    Jolo Pascual
    This document discusses microscopy techniques used to view bacteria. It describes the parts of a brightfield microscope and lists three types of microscopes used in bacteriology laboratories: brightfield, phase contrast, and fluorescence microscopes. Specific organisms that can be seen with each microscope are identified. Formulas for calculating actual size of organisms from their image size under different magnifications are provided, along with examples of using the formulas to determine image size and magnification.

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    W1BACT211LAB

    Uploaded by

    Jolo Pascual
    30 views2 pages
    This document discusses microscopy techniques used to view bacteria. It describes the parts of a brightfield microscope and lists three types of microscopes used in bacteriology laboratories: brightfield, phase contrast, and fluorescence microscopes. Specific organisms that can be seen with each microscope are identified. Formulas for calculating actual size of organisms from their image size under different magnifications are provided, along with examples of using the formulas to determine image size and magnification.

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    This document discusses microscopy techniques used to view bacteria. It describes the parts of a brightfield microscope and lists three types of microscopes used in bacteriology laboratories: brightfield, phase contrast, and fluorescence microscopes. Specific organisms that can be seen with each microscope are identified. Formulas for calculating actual size of organisms from their image size under different magnifications are provided, along with examples of using the formulas to determine image size and magnification.

    Copyright:

    © All Rights Reserved
    Download as DOCX, PDF, TXT or read online from Scribd
    Download as docx, pdf, or txt
    30 views2 pages

    W1BACT211LAB

    Uploaded by

    Jolo Pascual
    This document discusses microscopy techniques used to view bacteria. It describes the parts of a brightfield microscope and lists three types of microscopes used in bacteriology laboratories: brightfield, phase contrast, and fluorescence microscopes. Specific organisms that can be seen with each microscope are identified. Formulas for calculating actual size of organisms from their image size under different magnifications are provided, along with examples of using the formulas to determine image size and magnification.

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    PASCUAL, Jolo P.

    BACT211(LAB)
    BSMD 2Y2 - 7

    Week 1 Worksheet:
    Microscopy and Quantitation of Bacteria
    PART I Microscopy
     Enumerate all different parts of microscope and know its functions. (10pts)
    1. Framework - consists of the arm and the base; supports the whole microscope.
    2. Stage - platform supporting the microscopic slide.
     Mechanical stage - holds and moves the slide around
     Mechanical stage control - adjusts the position of the mechanical stage.
     Stage clip - holds the specimen in place.
     Stage hole - allows the passage of light.
    3. Light source - illuminates the specimen.
     Voltage regulator - adjusts the intensity of light.
     Neutral density filter - reduces intensity below lower limit allowed by the regulator/regulates
    intensity.
    4. Diopter adjustment ring - sharpens the image.
    5. Rotatable head - allows a choice of viewing positions.
    6. Lens systems
     Ocular/eyepiece - 10x magnification
     Objectives:
     Scanner - 4x
     Low Power Objective - 10x
     High Power Objective - 40x
     Oil Immersion Objective - 100x
     Condenser - collects and directs light from the illuminator to the slide being studied.
    7. Diaphragm/Iris Diaphragm - controls/regulates the amount of light that reaches the slide.
    8. Revolving nosepiece - where the objective lenses are attached, allows for easier switching.
    9. Focusing knobs
     Coarse adjustment knobs - initially brings the specimen into focus.
     Fine adjustment knobs – sharpens the focus.

    PART II Critical Thinking


    1. What is the principle of bright field microscopy? (1pt)
     In brightfield microscopy, illumination is the principle. Light is being absorbed by the stains, pigmentation, or
    dense areas of the specimen, producing contrast and allowing the viewer to see the specimen. The term
    “bright field” is derived from the dark specimen being contrasted by the surrounding bright viewing field.
    2. What are the different types of microscope used in the bacteriology lab? (2pts) 
     There are three types of microscopes being used in the bacteriology lab;
    1. Bright Field Microscope - examines the detail of the specimen through illumination.
    2. Phase Contrast Microscope - uses contrast enhancement techniques.
    3. Fluorescence Microscope - specimens are stained with fluorochrome or fluorescent dye.
    3. Identify what organism can be seen in different microscopy you have mentioned above? (2pts)
     These microscopes are used for different purposes; here are some examples
    1. Bright Field Microscope – bacteria and some organelles like peroxisomes and mitochondria can be
    seen
    2. Phase Contrast Microscopy – diverse group of protozoa, large flagellates and internal details of
    organisms that are unstained. Most living microscopic organisms are clearer in this microscopy.
    3. Fluorescence Microscopy – detailed observation of living organisms, particularly their internal
    structures. The bacteria Mycobacterium tuberculosis is an example of an organism that can be
    seen using fluorescence.

    Part III Computation (no formula, no point)


    1. What is the formula to determine the actual size of an organism? (1pt)
     The formula to determine the actual size of an organism is

    Actual size (um) = Image size (um)÷Magnification

    2. The actual size of an Escherichia coli  is 1.5um. What would be its image size if the magnification used is 1000X?
    (2pts)
    Image size(um) = Actual size (um) × Magnification
    =1.5um × 1000X
    Estimated size =1500um actual size

    3. What magnification is used if a Bacillus sp. have a microscopic size of 3.5um and an image size of 15mm? (2pts)
    Image size 15mm × 1000um = 15000um

    Magnification(X)= Image size(um) ÷ Actual size(um)


    =15000 um ÷ 3.5um
    =4285.71X

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