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Tutorial Test 1 Notes These study notes cover all the material included on tutorial test 1. They are concise and to the point to help reduce study time while still preparing effectively. Material is presented in the order in which they were introduced in

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Danton O' Day

used for examining living cellsMicroscopyless detail but more accurate new microscopic techniques are being can look 3D4Fluorescence Microscopydeveloped each year ranging from lightUV light is passed through based to UVbased to electronbasedtypes of microscopesfixedliving tissue to examine cellular fluorescence1Light Microscopyrequires 2 filtersnot general used for living cells1exciter only allows specific cellssections of fixed tissues are wavelength to excite sampleprepared stainedplaced on a 2barrier only allows specific slidewavelength of fluoresced used to examine cell shapelight through from sampleorganization of intracellular cells are tagged with fluorescent componentsmoleculesdont work for really small things can determine subcellular bc a single photon is bigger than localizationthe object itself can only say if if a fluorescentlylabelled object is thereantibody is used then called 2Phase Contrast Microscopyimmunofluorescence microscopygood for examining behaviour of DAPI a nuclear marker gets into living cellsgrooves of DNAfluoresceslight waves are shifted diffblue under UVparts of cell are seen at diff other fluorophores can be used levels of brightnessto mark proteinscan see general localization of can only see 1 fluorophore at a molecules large organellestime take picturesoverlap cellular constituents but need themstains to delineate what they are5Confocal Microscopygives more detail than DIC but uses laser light source instead of lots of artifacts UV more detailcompares 1 photon to the pinhole allows light from a photon next to it therefore certain plane to pass throughslight differences are blocks outoffocus planesexaggerated halo around clearer picturecellamplifies differences in no fuzzinessdiffractionsmaller pinholeclearer3Differential Interference Contrastbetteraka DICcan take images of all the uses a prismfilters to allow different slices of the samplecertain wavelengths of oriented computer will construct a 3D light through to highlight cellular imagestructures6TEM
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