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    Microscopes

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    David Robbie
    Microscopes use magnification through lenses or electron beams to observe microscopic organisms. Light microscopes use visible light and lenses to magnify up to 1000x, while electron microscopes use electron beams and can achieve over 100,000x magnification. Key principles of light microscopy include magnification through objective and ocular lenses, resolution of 0.2 μm allowing observation of bacterial but not viral structures, and various techniques like phase contrast, dark field, and fluorescence that increase contrast. Electron microscopes provide even higher resolution and magnification.

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    Microscopes

    Uploaded by

    David Robbie
    440 views13 pages
    Microscopes use magnification through lenses or electron beams to observe microscopic organisms. Light microscopes use visible light and lenses to magnify up to 1000x, while electron microscopes use electron beams and can achieve over 100,000x magnification. Key principles of light microscopy include magnification through objective and ocular lenses, resolution of 0.2 μm allowing observation of bacterial but not viral structures, and various techniques like phase contrast, dark field, and fluorescence that increase contrast. Electron microscopes provide even higher resolution and magnification.

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    Microscopes use magnification through lenses or electron beams to observe microscopic organisms. Light microscopes use visible light and lenses to magnify up to 1000x, while electron microscopes use electron beams and can achieve over 100,000x magnification. Key principles of light microscopy include magnification through objective and ocular lenses, resolution of 0.2 μm allowing observation of bacterial but not viral structures, and various techniques like phase contrast, dark field, and fluorescence that increase contrast. Electron microscopes provide even higher resolution and magnification.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as PPTX, PDF, TXT or read online from Scribd
    Download as pptx, pdf, or txt
    440 views13 pages

    Microscopes

    Uploaded by

    David Robbie
    Microscopes use magnification through lenses or electron beams to observe microscopic organisms. Light microscopes use visible light and lenses to magnify up to 1000x, while electron microscopes use electron beams and can achieve over 100,000x magnification. Key principles of light microscopy include magnification through objective and ocular lenses, resolution of 0.2 μm allowing observation of bacterial but not viral structures, and various techniques like phase contrast, dark field, and fluorescence that increase contrast. Electron microscopes provide even higher resolution and magnification.

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    Microscope Techniques Microscopes

    Microscopes
    Most important tool for studying microorganisms Use viable light to observe objects Magnify images approximately 1,000x
    Electron microscope, introduced in 1931, can magnify images in excess of 100,000x Scanning probe microscope, introduced in 1981, can view individual atoms

    Principles of Light Microscopy


    Light Microscopy
    Light passes through specimen, then through series of magnifying lenses Most common and easiest to use is the brightfield microscope Important factors in light microscopy include
    Magnification Resolution Contrast

    Principles of Light Microscopy


    Magnification
    Microscope has two magnifying lenses
    Called compound microscope Lens includes
    Ocular lens and objective lens Most bright field scopes have four magnifications of objective lenses, 4x, 10x, 40x and 100x

    Lenses combine to enlarge objects


    Magnification is equal to the factor of the ocular x the objective
    10x X 100x = 1,000x

    Principles of Light Microscopy


    Resolution
    Usefulness of microscope depends on its ability to clearly resolve two objects that are very close together
    Resolving power is defined as the minimum distance existing between two objects where those objects still appear as separate objects Resolving power determines how much detail can be seen

    Principles of Light Microscopy


    Resolution
    Resolution depends on the quality of lenses and wavelength of illuminating light (blue filter gives the best resolution)
    How much light is released from the lens

    Maximum resolving power of most brightfield microscopes is 0.2 m (1 um =1x10-6 M)


    This is sufficient to see most bacterial structures (>1um) Too low to see viruses (< 0.3 um)

    Principles of Light Microscopy


    Examples of light microscopes that increase contrast
    Phase-Contrast Microscope Interference Microscope Dark-Field Microscope Fluorescence Microscope Confocal Scanning Laser Microscope

    Principles of Light Microscopy


    Phase-Contrast
    Uses special lenses to amplify differences between refractive indexes of cells and surrounding medium Uses set of rings and diaphragms to achieve resolution

    Principles of Light Microscopy


    Dark-Field Microscope
    Reverse image
    Specimen appears bright on a dark background
    Like a photographic negative

    Achieves image through a modified condenser

    Principles of Light Microscopy


    Fluorescence Microscope
    Used to observe organisms that are naturally fluorescent or are flagged with fluorescent dye
    Fluorescent molecule absorbs ultraviolet light and emits visible light Image fluoresces on dark background

    Principles of Light Microscopy


    Electron Microscope
    Uses electromagnetic lenses, electrons and fluorescent screen to produce image Resolution increased 1,000 fold over brightfield microscope
    To about 0.3 nm (1 nm = 1x10-9 M)

    Magnification increased to 100,000x Two types of electron microscopes


    Transmission Scanning

    Principles of Light Microscopy


    Transmission Electron Microscope (TEM)
    Used to observe fine detail Directs beam of electrons at specimen
    Electrons pass through or scatter at surface
    Shows dark and light areas Darker areas more dense

    Specimen preparation through


    Thin sectioning Freeze fracturing or freeze etching

    Principles of Light Microscopy


    Scanning Electron Microscope (SEM)
    Used to observe surface detail Beam of electrons scan surface of specimen
    Specimen coated with metal
    Usually gold

    Electrons are released and reflected into viewing chamber Some atomic microscopes capable of seeing single atoms

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