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  • 1

    Microscope (Visual Observation) Experiments

    Prepared by Dr. C. S. Tritt September 25, 2007

  • 2

    Purpose Provide students with the opportunity to practice:

    the scientific method, use of literature resources record keeping (logbooks), report writing, visual observation and microscope use.

  • 3

    Specific Requirements

    See Microscope Experiment Specific Requirements handout.

  • 4

    M ic

    ro sc

    o p e

    Pa rt

    s –

    1 Objectives

    Stage

    Collimator

    Light source

  • 5

    M ic

    ro sc

    o p e

    Pa rt

    s –

    2

    Light Path Selection Rod

  • 6

    M ic

    ro sc

    o p e

    Pa rt

    s –

    3

    Focus Knobs

    Stage Positioning Knobs

  • 7

    M ic

    ro sc

    o p e

    Pa rt

    s –

    4

    Power Switches Power Connector

    USB Connector

  • 8

    Correctly Orientated Slide

  • 9

    Microscope Handling & Use

  • 10

    Microscopic Dimensions The following table compares the size of some common units used in microscopic work. 1 mm = 10-3 m = 103 μm 1 m = 106 μm or 1 μm = 10-6 m 1 nm = 10-9 m and 1 Å = 10-10 m 1 inch = 25,380 μm Typical human cells are 10 μm to 20 μm in diameter.

    Corrected 12/10/04

  • 11

    Objectives & Eyepieces The visual (with your eye) magnification of a microscope is the product of the objective (4, 10, 40 or 100 x) and eyepiece (10x) magnifications. When using the video camera in a microscope, only the objective magnification is relevant. Stating magnifications in terms of magnification (x) is meaningless once an image is captured.

  • 12

    Scale Bars (Calibration) It is critically important to place scale bars on photomicrographs do to the ease with which images can now be resized. The Motic microscope software includes features that will provide measures, but I prefer a manual approach.

  • 13

    Calibration Slide

  • 14

    4x Calibration Image

    1500 μm circle with 4x objective – as captured (1500umAt4x.png).

  • 15

    4x Image with Scale Bar

    1500 μm circle with 4x objective – with added text and scale bar.

  • 16

    40x Calibration Image

    Scale at 40x as captured (40xScaleWOBar.png)

  • 17

    40x Image with Bar

    Scale at 40x with Bar (40xScaleWBar.png)

  • 18

    Captured Image

    Kidney with 40x Objective (40xKidney.tif)

  • 19

    Captured Image w/Bar

    40x Kidney with Scale Bar (40xKidneyWBar.png)

  • 20

    Scale with 10x Objective

  • 21

    Scale with 100x Objective

  • 22

    Saving & Using Motic Images The default Motic image format (.img) can not be opened in other software. Save images in .tif or .jpg formats using File | Save as… Once saved, I generally process images as .jpg or .png files. See the Using Motic Software handout.

  • 23

    Image Editing Software Microsoft Paint and Photo Editor comes with your computer but are very limited in their features. I recommend you use ImageJ (see separate handout).

  • 24

    Power versus Resolution In past quarters, students have concluded they needed “higher power” microscopes. The 100x objectives on these microscope are about the practical limit for light microscope. What is generally needed is more resolution (or resolving power). See the examples on the next slide.

  • 25

    Resolution versus Power K id

    n ey

    a t

    4 0 x

    K id

    n ey

    a t

    4 0 x

    w it h

    re d u ce

    r es

    o lu

    ti o n

    (b lu

    rr ed

    e le

    ct ro

    n ic

    al ly

    )

    K id

    n ey

    a t

    4 0 x

    w it h

    en h an

    ce d

    el ec

    tr o n ic

    al ly

    K id

    n ey

    a t

    1 0 0 x

  • 26

    Ethical Enhancement There are a number of photo editing programs that allow enhancement (for example changing the brightness and contrast) of images. The degree to which scientific images should be enhanced is still a subject of debate. If you choose to enhance your images, be sure to disclose this fact in your report and caption.

  • 27

    Other Relevant Handouts

    Making and Scanning Sketches Using the ImageJ Photo Editor Inserting Figures into Word Documents Installing the Motic Microscope Imaging Software

    Microscope�(Visual Observation)�Experiments Purpose Specific Requirements Microscope Parts – 1 Microscope Parts – 2 Microscope Parts – 3 Microscope Parts – 4 Correctly Orientated Slide Microscope Handling & Use Microscopic Dimensions Objectives & Eyepieces Scale Bars (Calibration) Calibration Slide 4x Calibration Image 4x Image with Scale Bar 40x Calibration Image 40x Image with Bar Captured Image Captured Image w/Bar Scale with 10x Objective Scale with 100x Objective Saving & Using Motic Images Image Editing Software Power versus Resolution Resolution versus Power Ethical Enhancement Other Relevant Handouts

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