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Toxicity: Aome fluorochromes can be toxic to cells, to tissues, in vivo, or by producing mutations. [10] Limited resolving power: Fluorescence microscopes are limited in their ability to distinguish close objects at the macroscopic level. In comparison, electron microscopes for example, have the capacity to resolve at a much smaller range.
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.
A simplified Jablonski diagram illustrating the change of energy levels.. The principle behind fluorescence is that the fluorescent moiety contains electrons which can absorb a photon and briefly enter an excited state before either dispersing the energy non-radiatively or emitting it as a photon, but with a lower energy, i.e., at a longer wavelength (wavelength and energy are inversely ...
The fluorophore absorbs light energy of a specific wavelength and re-emits light at a longer wavelength. The absorbed wavelengths, energy transfer efficiency, and time before emission depend on both the fluorophore structure and its chemical environment, since the molecule in its excited state interacts with surrounding molecules.
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.
A mosaic constructed from a series of 53 images taken through three spectral filters by Galileo ' s imaging system as it flew over the northern regions of the Moon in December 1992. A false-color image from the Meteor M2-2 satellite's imager MSU-MR. The image was received by an amateur radio station and is derived from the HRPT data.
Historically, fluorophores such as fluorescein, rhodamine, Cy3 and Cy5 have been used in a wide variety of applications. These dyes have limitations for use in microscopy and other applications that require exposure to an intense light source such as a laser, because they photobleach quickly (however, bleaching can be reduced at least 10 fold using oxygen scavenging).
The following is a list of stars with resolved images, that is, stars whose images have been resolved beyond a point source. Aside from the Sun , observed from Earth , stars are exceedingly small in apparent size, requiring the use of special high-resolution equipment and techniques to image.