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Improving Plant Cell Analysis Requires Good TIRF

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Lynn Savage, Features Editor, [email protected]

Total internal reflection is the curious interaction of light and matter that makes fiber optic telecommunications possible, but its use in biological imaging may be even more interesting still. In any method of fluorescence microscopy, control over which particles fluoresce and which do not is a key challenge. For the most part, the struggle is to get the minimum amount of fluorophores to emit light – at just the right time – before snapping your subject’s photo. Too many particles glowing in the background, beyond the focal plane, create unwanted noise, obscuring the...Read full article

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    Published: October 2011
    Glossary
    critical angle
    The least angle of incidence at which total internal reflection takes place. The angle of incidence in a denser medium, at an interface between the denser and less dense medium, at which the light is refracted along the interface. When the critical angle is exceeded, the light is totally reflected back into the denser medium. The critical angle varies with the indices of refraction of the two media with the relationship: where Ic is the critical angle; n´ the refractive index of...
    fluorescence
    Fluorescence is a type of luminescence, which is the emission of light by a substance that has absorbed light or other electromagnetic radiation. Specifically, fluorescence involves the absorption of light at one wavelength and the subsequent re-emission of light at a longer wavelength. The emitted light occurs almost instantaneously and ceases when the excitation light source is removed. Key characteristics of fluorescence include: Excitation and emission wavelengths: Fluorescent materials...
    fluorescence microscopy
    Fluorescence microscopy is a specialized optical imaging technique used in biology, chemistry, and materials science to visualize and study specimens that exhibit fluorescence. Fluorescence is the phenomenon where a substance absorbs light at one wavelength and emits light at a longer wavelength. In fluorescence microscopy, fluorescent dyes or proteins are used to label specific structures or molecules within a sample. The basic principles of fluorescence microscopy involve illuminating the...
    actin filament bundlesAndor TechnologyArabidopsis thalianabackground noisebiological imagingBiophotonicsbotanybundling proteinscalcium-ion channelscamerasCatherine A. KonopkaChristopher J. StaigerCobolt ABCommunicationscritical anglecytoskeletonevanescence fieldFeaturesfluorescencefluorescence microscopyfluorophoresFranceGema Vizcay-Barrenagreen fluorescent proteinhamamatsuHeNe lasersillumination in microscopyImagingIndianalensesMelles GriotMicroscopymicrotubulesmirrorsNikonNorthern IrelandOlympusOpticsplant biologyproteinsPurdue Universityroot hairsRutherford Appleton LaboratorySebastian Y. Bednarekseedlingssingle-molecule microscopysubcritical anglesSwedenTIRFtotal internal reflection fluorescence microscopyU.K.Université Joseph FourierUniversity of NottinghamUniversity of Wisconsin MadisonVAEMvariable-angle epifluorescence microscopyvillinvillin1villin3WisconsinZeissZoe A. Wilson

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