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Histology Microscopy

Topic 2: Special Microscopes

Stereo Microscope 2 light microscopes combined Low magnification/Large focus More powerful than objective lens
Uses for stereo microscope Colposcope (Vaginal examination) Microsurgery Dissection (papillae) Parasitology and Pathology
Dark Field Microscope Cartoid filter: Blocks central light Object scatters light Object illuminated on a dark field
Uses for Dark Field Microscope Unstained organims Tracking movement of micro-organisms
Principle of Phase Contrast Microscope Varying thicknesses in object Varying refractive indices Microscope amplifies these variances
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
Uses of Phase Contrast Microscope Living cells Unstained cells Cell cultures
Nomarsky Microscope 2 Prisms Polarised light Prism 1: Splits light Split Light enters object close to eachother
Nomarsky Microscope (Prism 2) Prism 2 reassembles the light Contrast due to: Difference in refractive index not thickness
Uses of Nomarsky Microscope Cell cultures Unstained cells Living cells Immunostained material
Polarisation Microscope 2 filters Polarizator - Condensing lens Analizator - Objective lens
Polarizator & Analizator Filters light onto one plane Analizator is // : Field of vision is bright Analizator is perp.: Field of vision is dark
Anisotropic Structures that are able to rotate light to another plane. Shows up bright on a dark field eg. Crystals
Use of Polarisation Microscope Membrane analysis Diagnositics
Flouresence Microscope UV light source 2 filters pre and post specimen Specimen absorbs light, emits a light with bigger wavelength
Colour of self fluorescing tissues: Collagen Porifines Carotinids Lipofuscin Light Blue (Collagen) Red (Porfirines) Yellow (Carotinids/Lipofuscin)
Flourescent Dyes (F.A.R.) Flourescine Acridine Orange Rhodamine
Confocal Laser Scanning Microscope Laser beam excites molecules Laser moves across the object Light focused (w/pin hole) Records image Thicker objects
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)
Created by: 10fang711



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