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    Microscopy and Cytology

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    Kristine Fabellon
    This document provides a review of microscopy and cytology lab techniques. It covers: 1) The parts of the compound light microscope and principles of microscopy such as magnification, field of view, depth of focus, and resolving power. 2) Types of microscopes including the dissecting microscope. 3) Preparing wet mounts and examining eukaryotic cells like Elodea leaf, onion skin, and cheek cells. 4) Bacterial cell shapes and Gram staining of prokaryotic cells. 5) Models of animal cells, plant cells, and their organelles.

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    Microscopy and Cytology

    Uploaded by

    Kristine Fabellon
    103 views4 pages
    This document provides a review of microscopy and cytology lab techniques. It covers: 1) The parts of the compound light microscope and principles of microscopy such as magnification, field of view, depth of focus, and resolving power. 2) Types of microscopes including the dissecting microscope. 3) Preparing wet mounts and examining eukaryotic cells like Elodea leaf, onion skin, and cheek cells. 4) Bacterial cell shapes and Gram staining of prokaryotic cells. 5) Models of animal cells, plant cells, and their organelles.

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    Learn more about cells

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    This document provides a review of microscopy and cytology lab techniques. It covers: 1) The parts of the compound light microscope and principles of microscopy such as magnification, field of view, depth of focus, and resolving power. 2) Types of microscopes including the dissecting microscope. 3) Preparing wet mounts and examining eukaryotic cells like Elodea leaf, onion skin, and cheek cells. 4) Bacterial cell shapes and Gram staining of prokaryotic cells. 5) Models of animal cells, plant cells, and their organelles.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    103 views4 pages

    Microscopy and Cytology

    Uploaded by

    Kristine Fabellon
    This document provides a review of microscopy and cytology lab techniques. It covers: 1) The parts of the compound light microscope and principles of microscopy such as magnification, field of view, depth of focus, and resolving power. 2) Types of microscopes including the dissecting microscope. 3) Preparing wet mounts and examining eukaryotic cells like Elodea leaf, onion skin, and cheek cells. 4) Bacterial cell shapes and Gram staining of prokaryotic cells. 5) Models of animal cells, plant cells, and their organelles.

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    Unit 7: Review Microscopy and Cytology Lab

    Part I: Microscopy A. Parts of the compound light microscope


    A
    Examine the labeled microscope in your lab manual. Make sure you can identify all the parts.

    B C E

    F G H I J K L Continue Continue Side view Front view M N

    Part I: Microscopy B. Principles of microscopy


    Magnification the factor by which a specimen is enlarged. For each specimen viewed, the total magnification is calculated by multiplying the magnification of the ocular lens by the magnification of the objective. The light microscopes ocular magnification is 10X. Parfocal and parcentral imaging Parfocal imaging the ability of the microscope image to remain in focus when switching from one objective to the next. Fine adjustment using the fine adjustment knob might be needed, and the amount of light may need to be adjusted using the iris diaphragm lever and rheostat. Parcentral imaging the image will remain in the center of view when objective lenses are changed.

    Principles of microscopy continued

    Field of view
    Field of view the microscope field that appears as a circle when viewing a slide through the ocular and objective lenses. As the magnification is increased, the diameter of the field will decrease. You will need to determine the width of this circle so that you can make estimates about the size of an object observed. A ruler is placed across the stage. You are viewing the image through the scanning objective (4X). The diameter of the field is 4.2 mm. Convert to m. Using the above information you can calculate the field of view for the low power (10X), high power (40X), and oil immersion objectives (100X). You should also record area of your field of view.

    1 mm

    Continue

    Continue

    Principles of microscopy continued


    Depth of focus
    Depth of focus also known as depth of field. It is the vertical distance or thickness of an object that remains in focus of one time. A 3-colored thread slide was focused. Note that one thread is in focus and the others are not. The clearly focused thread lies on top of the mount.

    Principles of microscopy continued


    Image orientation
    Due to the optics involved with the light microscope, the image seen will be real, inverted, and magnified by the objective. The image is then magnified again by the ocular lens.

    Continue

    40X

    400X

    Continue

    40X

    Principles of microscopy continued


    Resolving power
    Resolving power the degree at which two adjacent points on a specimen are seen as separate detailed images.

    Part I: Microscopy C. Other types of microscopes

    Dissecting microscope
    The dissecting microscopes are useful when working with larger specimens. The large distance between the objective and the specimen allows for viewing thicker mounts and dissections. The illumination is from both above and below. Unlike the compound microscope, the image you see is not inverted.

    Contrast
    Contrast how well the details of a specimen stand out against a background. Stains and lighting are used to increase contrast to see detail. See the phase contrast micrograph in your lab manual.

    Continue

    Continue

    Part II: Cytology A. Preparing wet mounts


    To prepare a wet mount you need to a clean slide.

    Part II: Cytology B. Eukaryotic cells


    Elodea leaf wet mount

    A leaf from a sprig of Elodea is used as the specimen for making a wet mount.

    Cell wall Central vacuole location Cell membrane

    Add a drop of water onto the center of a microscope slide. Then take a thin sample of your specimen and place it in the drop of water.

    Examine the parts of the cell in this micrograph.

    Cytoplasm
    Add a cover slip to your prepared slide.

    Chloroplasts

    Continue

    Continue

    Eukaryotic cells continued


    Onion leaf cell wet mount
    A fleshy leaf from white onion was removed. The leaf was folded back to remove the thin layer of epidermis. A piece of the epidermis was placed on a microscope slide containing a drop of iodine. Examine the parts of the cell in this micrograph.

    Eukaryotic cells continued


    Cell membrane
    Human cheek cells wet mount The inside of the mouth was scraped with a toothpick. The scrapings were stirred in a drop of methylene blue stain. Examine the parts of the cell in this micrograph.

    Cell wall

    Cytoplasm Nucleus

    Cell membrane

    Cytoplasm

    Location of central vacuole Nucleolus

    Nucleus Continue

    Continue

    Part II: Cytology C. Prokaryotic cells


    Bacteria have diverse shapes. The most common bacterial shapes include coccus (cocci, pl.), bacillus (bacilli, pl.), and spirillum (spirilla, pl.). Bacteria are stained with Grams stain. Pink to red color indicate Gram negative and purple to blue color indicate Gram positive.

    Prokaryotic cells continued

    Bacillus Gram positive cocci

    Gram negative spirilla

    (The arrow shows an aggregate of about 3 bacilli.)

    Continue

    Continue

    Prokaryotic cells continued


    Coccus Spirillum

    Prokaryotic cells continued

    (You see clusters of spirilla in this view.)

    (The arrows are pointing to single cocci. You also see clusters of cocci at other areas.)

    Continue

    Continue

    Part II: Cytology D. Models


    C A Animal cell model #1 B
    A. Cell membrane B. Golgi apparatus C. Mitochondria D. Cytoplasm E. Endoplasmic reticulum F. Nucleus G. Pair of centrioles

    Models continued

    Animal cell/Plant cell Model #2


    A. Nuclear envelope B. Nucleolus C. DNA (chromatin) D. Nuclear pore

    D E G F

    B C

    Continue

    Continue

    Models continued
    A Plant cell Model #3
    A. Plasma membrane B. Mitochondria C. Golgi apparatus D. Cell wall E. Endoplasmic reticulum F. Cytoplasm G. Central vacuole H. Chloroplasts I. Nucleus

    B E F G C I H

    End of Lab Review

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