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Starting in 1994, light sheet fluorescence microscopy was developed as orthogonal plane fluorescence optical sectioning microscopy or tomography (OPFOS) [4] mainly for large samples and later as the selective/single plane illumination microscopy (SPIM) also with sub-cellular resolution. [5] This introduced an illumination scheme into ...
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.
Moiré Effect. Structured illumination microscopy (SIM) is a method of super-resolution microscopy which is performed by acquiring multiple images of the same sample under different patterns of illumination, then computationally combining these images to achieve a single reconstruction with up to 2x improvement over the diffraction limited lateral resolution.
Optical sectioning is the process by which a suitably designed microscope can produce clear images of focal planes deep within a thick sample. This is used to reduce the need for thin sectioning using instruments such as the microtome. Many different techniques for optical sectioning are used and several microscopy techniques are specifically ...
Critical illumination or Nelsonian illumination is a method of specimen illumination used for transmitted and reflected light (trans- and epi-illuminated) optical microscopy. Critical illumination focuses an image of a light source on to the specimen for bright illumination. Critical illumination generally has problems with evenness of ...
Bright-field microscopy (BF) is the simplest of all the optical microscopy illumination techniques. Sample illumination is transmitted (i.e., illuminated from below and observed from above) white light, and contrast in the sample is caused by attenuation of the transmitted light in dense areas of the sample.
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 ...
STED microscopy is one of several types of super resolution microscopy techniques that have recently been developed to bypass the diffraction limit of light microscopy to increase resolution. STED is a deterministic functional technique that exploits the non-linear response of fluorophores commonly used to label biological samples in order to ...