Methods and apparatus for optical imaging and scanning of holes machined, drilled or otherwise formed in a substrate made of composite or metallic material. The method utilizes an optical instrument for imaging and scanning a hole in combination with an image processor configured (e.g., programmed) to post-process the image data to generate one complete planarized image without conical optical distortion. The optical instrument includes an optical microscope with confocal illumination and a conical mirror axially positioned to produce a full 360-degree sub-image with conical distortion. In the post-processing step, a mathematical transformation in the form of computer-executable code is used to transform the raw conical sub-images to planar sub-images. The planarized sub-images may be stitched together to form a complete planarized image of the hole.

A compact single-axis confocal endomicroscope is provided, capable of complying within 2.8 mm diameter endoscope space requirements. The single-axis confocal endomicroscope uses a folded path design achieved between a fixed mirror and a lateral plane scanning mirror thereby producing a high numerical aperture that allows for diffraction-limited resolution with sub-surface depths. The scanning mirror is formed on a fixed-position, scanning MEMS assembly and has a central aperture that allows for illumination beam expansion in the folded path design. A series of spacers are used to retain beam focusing optical elements in fixed positioned relative to the scanning MEMS assembly for coupling with a single mode fiber.

A confocal microscope system includes a light source configured to form a light beam, a scanning unit, and an objective lens. The scanning unit is in the form of a mechanically driven scanning unit with a controllable scanning trajectory, and is configured to direct the light beam through the scanning trajectory. The objective lens defines a pupil plane and a focal plane. The light beam is directed from the scanning unit to the objective lens. The confocal microscope system is configured for multi-color line-scanning confocal microscopy, and implements multi-color fluorescence imaging without laser excitation crosstalk.

The invention relates to an apparatus for manipulating a focus of excitation light on or in a sample, particularly in a microscope, comprising a light source for emitting excitation light, an excitation beam path for guiding the excitation light onto or into the sample, the excitation beam path comprising an objective for guiding the excitation light onto or into the sample and a wavefront modulator for modulating the excitation light, and a control device for driving the wavefront modulator. According to the invention, the apparatus is characterized in that the control device is designed for driving the wavefront modulator to generate a number of shaped waves on or in the sample, that a focus is generated at a specified location on or in the sample by superposition of the shaped waves and that, for manipulating the location of the focus on or in the sample, a device for imposing variably stepped phase shifts upon the shaped waves is present, where the phase shifts imposed in each case on the shaped waves change stepwise between different shaped waves. In further aspects, the invention relates to microscope and a method for manipulating a focus of excitation light on or in a sample.

A compact single-axis confocal endomicroscope is provided, capable of complying with 2.8 mm diameter endoscope space requirements. The single-axis confocal endomicroscope uses a folded path design achieved between a fixed mirror and a lateral plane scanning mirror thereby producing high numerical apertures that allow for diffraction-limited resolution in sub-surface scanning. The scanning mirror has a central aperture that allows for illumination beam expansion in the folded path design.

A confocal microscope unit according to an embodiment includes: a first subunit which includes a light source, a pinhole plate, and a photodetector; a second subunit which includes a light source, a pinhole plate, and a photodetector; a scan mirror which scans excitation light on a sample and guides fluorescence generated from the sample to the first and second subunits; a scan lens which guides the excitation light and guides the fluorescence to the scan mirror; and a main housing which is attachable to a connection port and to which the scan mirror, the scan lens, and the subunits are fixed, wherein the first subunit includes a dichroic mirror that separates the excitation light and fluorescence handled by the own unit from those handled by the second subunit.

Embodiments are for a confocal microscope unit attached to a connection port of a microscope including: light sources which output irradiation light to a sample to be observed; photodetectors which detect observation light generated from a sample in response to the irradiation light; a scan mirror which scans the irradiation light on the sample and guides the observation light generated from the sample to the photodetectors; a scan lens which guides the irradiation light scanned by the scan mirror to the microscope optical system and guides the observation light focused by the microscope optical system to the scan mirror; a lens barrel to which the scan lens is fixed; an attachment portion which attaches the lens barrel to the connection port; and a movable portion which supports the lens barrel so that an angle of the lens barrel with respect to the attachment portion is changeable.

High speed laser scanning systems and methods are disclosed for imaging target samples. Embodiments of the laser scanning systems and methods include separating a laser beam into two or more different pathways and pulsing light along the two or more pathways. The different pathways intersect the surface of a moving mirror at different locations and light traveling in a pathway that is being approached by an edge of the moving reflective surface is turned off while light traveling in a different pathway that is farther from the edge of the moving reflective surface is turned on. Embodiments include reflective surfaces that form a rotating polygonal scanner. Further embodiments include recombining the two or more different pathways, focusing the laser light on a test sample, and imaging the light reflected from the test sample.

The field of view (FOV) of a nonlinear optical microscope (NLOM) is expected to be large enough for employing high-speed raster scanning on a mesoscale volumetric biological sample. Concurrently, three-dimensional (3D) visualization of fine sub-micron biological structures requires high enough lateral and axial resolutions, enforcing a high numerical aperture (NA) objective lens to be employed, thereby limiting the FOV of an NLOM. The invention is directed to a laser scanning NLOM, or to a large-angle optical raster scanning system, for deep biological tissue imaging with a large FOV of more than one square millimeter, up to 1.6×1.6 mm.sup.2, while simultaneously maintaining a sub-femtoliter effective 3D resolution by means of a high-NA and low magnification objective lens and further maintaining a high acquisition speed with synchronized sampling, limited by the repetition rate of a high repetition rate pulsed laser source, thereby exceeding Nyquist Criterion for resolving micro-optical resolution throughout a horizontal FOV of more than one millimeter.

Disclosed are a device and method for measuring fluorescence signal in multi-illumination mode and use of the method. An excitation light is modulated on an illumination excitation light path, to generates excitation illumination patterns at different phases on a sample after passing through an objective lens; a high-speed switching device is arranged on a fluorescence collection light path to switch the position of a fluorescence image of the sample on a target plane of a photoelectric sensor, and a plurality of sub-images can be simultaneously obtained after one exposure through synchronous operation of multi-illumination excitation light paths and fluorescence imaging light paths, which correspond to fluorescence signals in a plurality of illumination modes.