Figure 3.4 Approximate size of various types of cells. ~10 um. Red Blood Cells = mm 1500 um. Width of penny Pearson Education, Inc.
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1 Figure 3.4 Approximate size of various types of cells. ~10 um Red Blood Cells 1.5mm 1500 um Width of penny = 1500
2 Figure 4.3 The limits of resolution (and some representative objects within those ranges) of the human eye and of various types of microscopes.
3 Official definitions Magnification, the ratio of an object s image size to its real size Resolution, the measure of the clarity of the image, or the minimum distance of two distinguishable points Contrast, visible differences in brightness between parts of the sample
4 Microscopy General Principles of Microscopy Magnification (enlarge) Resolution (tell apart 2 objects close together) Contrast (Differences in intensity between two objects)
5 General rule for any microscope/detector Smaller the wavelength smaller the object you can see (small objects need small hands) Think about giant fingers and texting
6 Figure 4.1 The electromagnetic spectrum. Smaller the object = Use smaller wavelength VIBGYOR
7 Microscopy Microscopes are used to visualize cells In a light microscope (LM), visible light is passed through a specimen and then through glass lenses Glass lenses focuses light and enlarges and resolves objects
8 Lenses refract (bend) the light, so that the image is magnified Light Air Glass Magnification Magnify using = 50/5 lenses = 10X Focal point 5 50 Specimen Convex lens Inverted, reversed, and Enlarged image
9 Microscopy Light Microscopy Bright-field microscopes Simple single lense Contain a single magnifying lens Similar to magnifying glass Leeuwenhoek used simple microscope to observe microorganisms
10 Figure 4.4 A bright-field, compound light microscope. occular lens Line of vision Ocular lens Remagnifies the image formed by the objective lens Body Transmits the image from the objective lens to the ocular lens using prisms Arm Objective lenses Primary lenses that magnify the specimen Stage Holds the microscope slide in position Condenser Focuses light through specimen Diaphragm Controls the amount of light entering the condenser Illuminator Light source Ocular lens Path of light Prism Body Objective lenses Specimen Condenser lenses Illuminator Coarse focusing knob Moves the stage up and down to focus the image Fine focusing knob Base objective lens
11 Microscopy Light Microscopy Bright-field microscopes Compound multiple lenses More than one - Series of lenses for magnification Light passes through specimen into objective lens Have one or two ocular lenses Total magnification = magnification of objective lens X magnification of ocular lens E.g. 10 times X 10 times = 100 times
12 Figure 4.5 The effect of immersion oil on resolution. Special type of objective lens = oil immersion lens Microscope objective Lenses Microscope objective Refracted light rays lost to lens Glass cover slip More light enters lens Glass cover slip Immersion oil Slide Slide Specimen Light source Light source Without immersion oil With immersion oil Increases magnification and resolution
13 Microscopy General Principles of Microscopy Contrast Differences in intensity between two objects, or an object and its background Increase contrast by staining
14 Figure 6.3ab 50 µm Brightfield (unstained specimen) Brightfield (stained specimen) Staining increases contrast
15 Figure 4.16 Simple stains.
16 paramecium
17 penicillium
18 spirogyra
19
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