Topic 2: Special Microscopes
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| Stereo Microscope | 2 light microscopes combined
Low magnification/Large focus
More powerful than objective lens
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| Uses for stereo microscope | Colposcope (Vaginal examination)
Microsurgery
Dissection (papillae)
Parasitology and Pathology
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| Dark Field Microscope | Cartoid filter: Blocks central light
Object scatters light
Object illuminated on a dark field
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| Uses for Dark Field Microscope | Unstained organims
Tracking movement of micro-organisms
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| Principle of Phase Contrast Microscope | Varying thicknesses in object
Varying refractive indices
Microscope amplifies these variances
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| Theory of Phase Contrast Microscope | Annulus diaphragm - Over condenser
Ring of light allowed through
Phase Ring - In objective lens
Changes wavelength by 0.25 (lambda)
Combines refracted and diffracted light
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| Uses of Phase Contrast Microscope | Living cells
Unstained cells
Cell cultures
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| Nomarsky Microscope | 2 Prisms
Polarised light
Prism 1: Splits light
Split Light enters object close to eachother
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| Nomarsky Microscope (Prism 2) | Prism 2 reassembles the light
Contrast due to:
Difference in refractive index not thickness
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| Uses of Nomarsky Microscope | Cell cultures
Unstained cells
Living cells
Immunostained material
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| Polarisation Microscope | 2 filters
Polarizator - Condensing lens
Analizator - Objective lens
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| Polarizator & Analizator | Filters light onto one plane
Analizator is // : Field of vision is bright
Analizator is perp.: Field of vision is dark
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| Anisotropic | Structures that are able to rotate light to another plane.
Shows up bright on a dark field
eg. Crystals
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| Use of Polarisation Microscope | Membrane analysis
Diagnositics
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| Flouresence Microscope | UV light source
2 filters pre and post specimen
Specimen absorbs light, emits a light with bigger wavelength
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| Colour of self fluorescing tissues: Collagen Porifines Carotinids Lipofuscin |
Light Blue (Collagen)
Red (Porfirines)
Yellow (Carotinids/Lipofuscin)
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| Flourescent Dyes (F.A.R.) | Flourescine
Acridine Orange
Rhodamine
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| Confocal Laser Scanning Microscope | Laser beam excites molecules
Laser moves across the object
Light focused (w/pin hole)
Records image
Thicker objects
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| Light beams in Flouresence Microscopy | Barrier 1: 450-490 nm
Barrier 2: <510 nm (reflected)
> 510 nm (pass)
Barrier 3: 520 to 560 nm (pass through)
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