A method and associated apparatus for generating instantaneous and uniform total internal reflection fluorescence (TIRF) excitation. An annular fiber bundle and is used with spatially incoherent light to provide appropriate illumination matched to parameters of the back focal plane of an oil-immersion or in-air imaging objective lens, enabling quantitative shadowless TIRF imaging.

Provided are a method and apparatus for imaging a self-luminous object on a biological sample film (A); said imaging apparatus comprises a housing (1), an optical-electrical conversion element (2), and an image correction device (3); the inside of the housing (1) constitutes a darkroom space; the optical-electrical conversion element (2) is arranged in the housing (1); the optical-electrical conversion element (2) is used, when the biological sample film (A) is not inserted, for obtaining a first dark field image inside the housing (1); after affixing the biological sample film (A) to the surface of the optical-electrical conversion element (2), the optical-electrical conversion element (2) is also used for obtaining a second dark field image inside the housing (1); the image correction device (3) is used for correcting the second dark field image according to the first dark field image to obtain a target image corresponding to the self-luminous object. The imaging apparatus and imaging method can effectively and more accurately obtain a higher-definition target image of the self-luminous object; the operation process is simple and the time required for imaging is short; furthermore, the imaging apparatus has the advantages of a small structure, low cost of manufacture, convenient operation, and portability.

Provided are a method and apparatus for imaging a self-luminous object on a biological sample film (A); said imaging apparatus comprises a housing (1), an optical-electrical conversion element (2), and an image correction device (3); the inside of the housing (1) constitutes a darkroom space; the optical-electrical conversion element (2) is arranged in the housing (1); the optical-electrical conversion element (2) is used, when the biological sample film (A) is not inserted, for obtaining a first dark field image inside the housing (1); after affixing the biological sample film (A) to the surface of the optical-electrical conversion element (2), the optical-electrical conversion element (2) is also used for obtaining a second dark field image inside the housing (1); the image correction device (3) is used for correcting the second dark field image according to the first dark field image to obtain a target image corresponding to the self-luminous object. The imaging apparatus and imaging method can effectively and more accurately obtain a higher-definition target image of the self-luminous object; the operation process is simple and the time required for imaging is short; furthermore, the imaging apparatus has the advantages of a small structure, low cost of manufacture, convenient operation, and portability.

A dark field illuminator for microscopic imaging is provided. The dark field illuminator is arranged above an adjustable lens group of a unit microscopic imaging module and corresponds to the adjustable lens group, a surface of the dark field illuminator is attached to a back of a sample slide, and the sample slide is located between the dark field illuminator and the adjustable lens group; the dark field illuminator includes a bright and dark field substrate and a dark field black background patch, the size of the dark field black background patch matches with that of the adjustable lens group, and the dark field black background patch is arranged close to or away from the adjustable lens group relatively to the bright and dark field substrate. Preferably, the bright and dark field substrate further has a recessed structure with a white diffuse reflection surface.

A dark field illuminator for microscopic imaging is provided. The dark field illuminator is arranged above an adjustable lens group of a unit microscopic imaging module and corresponds to the adjustable lens group, a surface of the dark field illuminator is attached to a back of a sample slide, and the sample slide is located between the dark field illuminator and the adjustable lens group; the dark field illuminator includes a bright and dark field substrate and a dark field black background patch, the size of the dark field black background patch matches with that of the adjustable lens group, and the dark field black background patch is arranged close to or away from the adjustable lens group relatively to the bright and dark field substrate. Preferably, the bright and dark field substrate further has a recessed structure with a white diffuse reflection surface.

A system in an embodiment can comprise an optical assembly, an surface-plasmon-resonance (SPR) light source, and an SPR camera. The optical assembly can comprise a hemispherical prism comprising a top surface configured to support a SPR sensor; and a high numerical aperture (NA) lens located distal from the top surface of the hemispherical prism. The SPR light source can be configured to emit a light beam for SPR imaging. The SPR camera can be configured to capture an SPR image. The SPR sensor further can comprise a surface configured to contact a sample. The high NA lens can be configured to refract the light beam toward the hemispherical prism. The hemispherical prism can be configured to collimate the light beam, as refracted by the high NA lens, toward the SPR sensor. The high NA lens further can be configured to receive and refract the light beam toward the SPR camera, after the light beam is reflected by the surface of the SPR sensor. Other embodiments are disclosed.

An illumination module may include a laser diode array configured to emit laser radiation; a phosphor illumination unit that is configured to emit phosphor radiation following an exposure to the laser radiation; a multiple-angle illumination unit; and intermediate optics that is configured to convey the phosphor radiation to the multiple-angle illumination unit. The multiple-angle illumination unit is configured to receive the phosphor radiation and to dark field illuminate a region of a sample wafer from multiple angles during inspection of the wafer.

Methods for imaging a sample using fluorescence microscopy, systems for imaging a sample using fluorescence microscopy, and illumination systems for fluorescence microscopes. In some examples, a method includes positioning the sample such that a plane of interest of the sample is coplanar with a focal plane of a detection objective of a microscope. The method includes positioning a paraboloidal mirror around the sample such that a focal point of the paraboloidal mirror is coplanar with the focal plane of the detection objective and the plane of interest of the sample. The method includes directing a beam of annularly collimated excitation light on the paraboloidal mirror to focus a disc of light on the sample and thereby to provide 360 degree lateral illumination of the sample. The method includes imaging the sample through the detection objective.

The invention relates to an illumination apparatus for a microscope, a microscope and a method for operating the illumination apparatus. The illumination apparatus has a sample space for holding a sample that is to be illuminated, and at least one laser light source. An objective for the directional emission of laser radiation of a first wavelength along a first optical axis that is directed into the sample space, and with a cover of the sample space by which the sample space is delimited at least on one of its sides. The cover further has a layer that is either impenetrable for the laser radiation over a blocking angle range of the illumination angle and is transmissive for radiation of a second wavelength over a transmitted light angle range, or has a controllable layer that, in a first control state, is transparent for radiation of the second wavelength and, in a second control state, is impenetrable for the laser radiation of the first wavelength.

A SPIM-microscope (Selective Plane Imaging Microscope) having a y-direction illumination light source and a z-direction detection light camera. An x-scanner generates a sequential light sheet by scanning the illumination light beam in the x-direction. The SPIM-microscope has an illumination optics having a zoom optics provided in a beam path of the illumination light beam, the zoom optics being adapted to change the focal length of the illumination light beam and adapted to detect a larger area of the object by sequentially detecting sequences of images along the y-direction that have an increased resolution along the z-direction. An image processing unit combines these sequences of images by image stitching into one large overall image.