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Microscopy
Maurice WetherallUniversity Senior College
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“This material has been developed as a part of the Australian School Innovation in Science, Technology and Mathematics Project funded by the Australian Government Department of Education, Science and Training as a part of the Boosting Innovation in Science, Technology and Mathematics Teaching (BISTMT) Programme.”
Microscopy
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Microscopes
• Our knowledge and understanding of the structure of cells has only been possible as a result of the use of microscopes.
• Light microscopes were first used in the 16th century.
• The electron microscope was developed in the 1930’s.
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Anton Van Leeuwenhoek’s Simple Microscope - 1670
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An English Tripod Microscope from around
1680
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A 19th
CenturyMicroscope
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A Current Compound
(Light) Microscope
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Image From Light Microscope:
Onion Root Cells X1000
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Compound Light Microscope
• Light from a light source is directed through the specimen.
• A combination of lenses is used to increase the resolving power of the human eye up to 500 times.
• Resolving power (or resolution) refers to the ability to distinguish fine detail.
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• The magnifying power is calculated by multiplying the individual powers of the eye-piece (ocular) and objective lenses.
• e.g. 10x eye-piece, 40x objective = 400x
• Field diameter is the actual distance across the field of view.
• As magnifying power increases the field diameter decreases proportionally.
Compound Light Microscope
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• The amount of light passing through the specimen can be adjusted using the iris diaphragm. This changes the contrast.
• The condenser lens is used to concentrate light on the specimen.
Compound Light Microscope
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• The following is a short movie of a living amoeba magnified 1000X through a compound light microscope
• An amoeba is a microscopic unicellular animal that lives in water
• Note the cytoplasmic movement, and also the way the whole cell moves.
Amoeba X1000
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Amoeba X1000
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• This is a special type of light microscope, which provides greater contrast.
• Structures, which could not usually be seen without staining, show up.
• Staining cells kills the cells.• Phase contrast microscopes can be used to
observe living cells.• e.g. movement of chromosomes during
mitosis can be viewed.
Phase Contrast Microscope
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• In an electron microscope, beams of electrons are focussed using magnetic lenses.
• The resolving power produced is up to 500,000 times greater than the human eye.
• Because a vacuum is needed, tissue has to be specially prepared, and so living cells cannot be examined.
Transmission Electron Microscope (TEM)
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Transmission Electron Microscope (TEM)
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Mitochondria
• The following slide shows mitochondria viewed through a transmission EM
• The internal structures of the mitochondria are visible. They are not visible through Light Microscopes.
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Rough Endoplasmic Reticulum
• The following slide shows a section of rough endoplasmic reticulum.
• These flattened membranes are involved in synthesis and transport of proteins in cells.
• The small, dark dots are ribosomes.
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Smooth Endoplasmic Reticulum
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Electron Micrograph of Golgi Body
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Scanning Electron Microscope
• An electron beam scans the surface of the specimen,
• Electrons are reflected off, and collected by a special electron detector.
• This provides an image, which appears on a computer screen and gives an impression of the outer shape of the specimen.
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Scanning Electron Microscope at Adelaide Microscopy
• The following slide shows Year 11 students using one of the scanning EMs at Adelaide Microscopy.
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Human Hair in a Scanning Electron
Microscope• The following specimens show human
hair, the first one is hair in good condition and is from a person using shampoo and conditioner that are free of Sodium Laurel Sulfate and Propylene Glycol
• The second is of poor quality hair from a person using shampoo and conditioner containing Sodium Laurel Sulfate and Propylene Glycol
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The End