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Schematic of a fluorescence microscope. The majority of fluorescence microscopes, especially those used in the life sciences, are of the epifluorescence design shown in the diagram. Light of the excitation wavelength illuminates the specimen through the objective lens.
Light microscopes are used for viewing stained samples at high magnification, typically using bright-field or epi-fluorescence illumination. Staining is not limited to only biological materials, since it can also be used to study the structure of other materials; for example, the lamellar structures of semi-crystalline polymers or the domain ...
A total internal reflection fluorescence microscope (TIRFM) is a type of microscope with which a thin region of a specimen, usually less than 200 nanometers can be observed. TIRFM is an imaging modality which uses the excitation of fluorescent cells in a thin optical specimen section that is supported on a glass slide.
Some of the recently developed super-resolution fluorescent microscope methods include stimulated emission depletion microscopy, saturated structured-illumination microscopy (SSIM), fluorescence photoactivation localization microscopy (FPALM), and stochastic optical reconstruction microscopy (STORM).
Fluorescence-lifetime imaging microscopy or FLIM is an imaging technique based on the differences in the exponential decay rate of the photon emission of a fluorophore from a sample. It can be used as an imaging technique in confocal microscopy , two-photon excitation microscopy , and multiphoton tomography.
Two-photon excitation microscopy of mouse intestine.Red: actin.Green: cell nuclei.Blue: mucus of goblet cells.Obtained at 780 nm using a Ti-sapphire laser.. Two-photon excitation microscopy (TPEF or 2PEF) is a fluorescence imaging technique that is particularly well-suited to image scattering living tissue of up to about one millimeter in thickness.
Light sheet fluorescence microscopy (LSFM) is a fluorescence microscopy technique with an intermediate-to-high [1] optical resolution, but good optical sectioning capabilities and high speed. In contrast to epifluorescence microscopy only a thin slice (usually a few hundred nanometers to a few micrometers) of the sample is illuminated ...
Light sheet fluorescence microscopy or selective plane imaging microscopy (SPIM) uses illumination that is done perpendicularly to the direction of observation, by using a thin sheet of (laser) light. Under certain conditions, this illumination principle can be combined with fluorescence correlation spectroscopy, to allow spatially resolved ...