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Lecture 9 - EXAM VERSION.docx

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Michael Jonz

Lecture 9EXAM VERSIONNovember21121211 PM RESEARCH TECNIQUESLight MicroscopeUses basic light path lightspecimen eyeImage inverted through slide and to your eye objective lensolight from top lenses on bottom thin prep inverted microLive or fixed cells and tissueoLight travels easily through cells and thick tissueoTissue upright microscopeoIsolated cells inverted microscopelight focused on specimen by condenser lenses1objective and eye piece lenses focus an image in eye1Phase of light changes when light passes through a living cell 1light passing through a thickdense part is retarted1TypesBright fieldTransmitted lightImage obtained by simple light transmission through a cell in culturePhase contrastConverts phase differences into changes in brightnessExploits wave propertiesoChange when entering and exiting celloexploits interference effects produced when 2 sets of waves recombineoexploiting shift creates contrastObserves phase shift through unstained cellsDifferential interference contrastSimilar to phase contrast but more definitionExploits interference effectsInterferes with light passing through cell instead of exploiting phase shiftLight is separated before interaction then put back together by prismsDark fieldRays of light directed from side lateral light source so that only scattered light enters microscope lensis shownCell appears bright against dark backgroundNote Change in brightnesscontrast can see shading contrast and textureChanging phase any light traveling through specimen interactionchange in phase uses wave properties PhaseDIC ComparedPhase contrast uses phase plate utilizes phase changesDIC prism used to interfere with light before entering specimenNote Direct lightunaltered not scattered light
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