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Lecture 10 - EXAM VERSION.docx

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Michael Jonz

Lecture 10EXAM VERSIONNovember22121139 PM Electron MicroscopyTransmission electron microscope TEMScanning electron microscope SEMLight source bombardment of electronsLens electron gun and magnetEffect high resolutionAdvantage resolution 2000X than light microscopeResolves fine structure of cellDisadvantage time consuming prep appropriate technique for some experiments Transmission EMLight source bombardment of electronsLens electron gunUse inside cellsEffect contrast dense imageMechanism Tissue cut and bombarded with electrons some pass through image some bounceCompared to LMSimilar to light but biggerFocuses electrons released from a gun onto specimen instead of using condenser to focus lightProcessElectrons emitted at filament or cathode source of light at top of cylindrical 1columnAccelerated by high voltage in vacuum by air filament by anode1pass through hole to form electron beam that travels down column2Magnetic coils focus electron beam Specimen stained with electron dense material produces electron dense 1imagesElectrons passing through are scattered by stained structuresthe rest are focused to form an imageNote Can increase density with reagentsElectron denseblack electrons scattered away that dont make it to detectorElectron lucent light areas where electrons were allowed to pass through forms image TEM Preptakes days or weeksTissue fixed in glutaraldehyde1Fixative adds densityAddition of OsO4 increases electron density1Dehydration and infiltration with plastic resin 1
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