Microscopes: LM (Video Microscopy and Image Processing, Super-Res Fluorescence Microscopy)

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Microscopes: LM (Video Microscopy and Image Processing, Super-Res Fluorescence Microscopy)
2013-09-02 09:30:00
Cell Bio

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  1. a.      Video Microscopy and Image Processing
                                                                  i.      some video cameras (like charge-coupled device, or CCD cameras) are made to be __, allowing for imaging at __ (good for __and/ or __ specimen)
                                                                ii.      video cameras can detect and amplify __ within a species so small objects are visible
    • very sensitive to light
    • low illuminations
    • live 
    • fluorescently stained
    • small differences in contrast
  2.                                                               i.      electronic images produced are readily converted to __ with a discrete number of picture elements each of which has an assigned __ and __corresponding to that site in the original image
                                                                ii.      can be stored on pc and edited
    • digital images
    • color and brightness value
  3. Laser Scanning Confocal Microscopy
    - disadvantage of LM
                                                                  i.      different depths of specimen reduce ability of forming a crisp image because those parts interfere with light rays from the plane in focus
  4.                                                               i.      1950: __ invented the __ that produces an image of a __                                                            ii.      specimen is illuminated by a finely focused __ that rapidly scans across the specimen at a __, thus illuminating only a __ within the specimen
    • Marvin Minsky
    • confocal microscope
    • thin plane situated within a much thicker specimen
    • laser beam
    • single depth
    • thin plane
  5.                                                               i.      used with __
    1.      after __at longer wavelength, light from the specimen is brought to focus at a site within the microscope that contains a __ (which, along with the plane are __)
    light rays from the illuminated plane  pass through the aperture, but other rays cannot
    • fluorescent optics
    • reemittence 
    • pinhole aperture
    • confocal
  6. a.      Super-Resolution Fluorescence Microscopy
                                                                  i.      scientists have been able to localize proteins at resolutions in the tens of nm  due to discovery of __
    1.      led to development of __, whose fluorescence emission at a certain wavelength can be switched on and off 
    • certain mutation of GFP polypeptide, which is nonfluorescent until activated by violet light
    • photoswitchable proteins
  7.                                                               i.      STORM (stochastic optical reconstruction microscopy)
    1.      uses __ and allows investigators to localize a single __molecule within a resolution < 20 nm with pulses of light of appropriate wavelengths
    • photoswitchable fluorescent proteins
    • fluorescent
  8. 1.      most labeled molecules remain dark, but some are __ and appear as __

    2.      the center can be determined with high accuracy
    • activated
    • spots hundreds nm wide
  9. 1.      process is repeated for more __, producing a lot of coordinates that represent the locationsof many of the __in the specimen
    2.      analysis leads to  __
    • imaging cycles
    • fluorophores 
    • reconstruction a super-resolution image of the field of view