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[1] [2] A fluorescence microscope is any microscope that uses fluorescence to generate an image, whether it is a simple set up like an epifluorescence microscope or a more complicated design such as a confocal microscope, which uses optical sectioning to get better resolution of the fluorescence image. [3]
In most light sheet fluorescence microscopes the detection objective and sometimes also the excitation objective are fully immersed in the sample buffer, so usually the sample and excitation/detection optics are embedded into a buffer-filled sample chamber, which can also be used to control the environmental conditions (temperature, carbon ...
Köhler illumination is a method of specimen illumination used for transmitted and reflected light (trans- and epi-illuminated) optical microscopy.Köhler illumination acts to generate an even illumination of the sample and ensures that an image of the illumination source (for example a halogen lamp filament) is not visible in the resulting image.
Diagram illustrating the light path through a dark-field microscope. The steps are illustrated in the figure where an inverted microscope is used. Light enters the microscope for illumination of the sample. A specially sized disc, the patch stop (see figure), blocks some light from the light source, leaving an outer ring of illumination. A wide ...
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.
IF can additionally be used in combination with other, non-antibody methods of fluorescent staining, e.g., the use of DAPI to label DNA. [10] [11] Examination of immunofluorescence specimens can be conducted utilizing various microscope configurations, including the epifluorescence microscope, confocal microscope, and widefield microscope. [12]
Most excitation filters select light of relatively short wavelengths from an excitation light source, as only those wavelengths would carry enough energy to cause the object the microscope is examining to fluoresce sufficiently. [1] The excitation filters used may come in two main types — short pass filters and band pass filters. [2]
The light path of a bright-field microscope is extremely simple; no additional components are required beyond the normal light-microscope setup. The light path begins at the illuminator or the light source on the base of the microscope. Often a halogen lamp is used. The light travels through the objective lens into the ocular lens, through ...