A spectral imaging device (12) includes an image sensor (28), a tunable light source (14), an optical assembly (17), and a control system (30). The optical assembly (17) includes a first refractive element (24A) and a second refractive element (24B) that are spaced apart from one another by a first separation distance. The refractive elements (24A) (24B) have an element optical thickness and a Fourier space component of the optical frequency dependent transmittance function. Further, the element optical thickness of each refractive element (24A) (24B) and the first separation distance are set such that the Fourier space components of the optical frequency dependent transmittance function of each refractive element (24A) (24B) fall outside a Fourier space measurement passband.

A fluorescence scanning microscope includes excitation and de-excitation light sources, which are designed to generate an excitation and a de-excitation light distribution, respectively. An illumination unit combines the light distributions to form a light distribution scanning over multiple illumination target points of a sample in such a way that an intensity maximum of the excitation light distribution and an intensity minimum of the de-excitation light distribution are spatially superimposed on one another. A detector detects fluorescence photons emitted from the respective illumination target point as a function of their arrival times. A processor evaluates the fluorescence photons with respect to the arrival times, generates a first pixel and a second pixel based thereon, assembles the first and second pixels to form first and second sample images, respectively, and, by means of the two sample images, determines a spatial offset between the intensity maximum and the intensity minimum.

A chromatic range sensor (CRS) system is provided that determines a workpiece thickness and includes an optical pen, an illumination source, a wavelength detector and a processing portion. The optical pen includes an optics portion providing axial chromatic dispersion, the illumination source is configured to generate multi-wavelength light and the wavelength detector includes a plurality of pixels distributed along a measurement axis. In operation, the optical pen inputs a spectral profile from the illumination source and outputs corresponding radiation to first and second workpiece surfaces of a workpiece (e.g., which may be transparent) and outputs reflected radiation to the wavelength detector which provides output spectral profile data. The processing portion processes the output spectral profile data to determine a thickness of the workpiece. In various implementations, the processing to determine the thickness may not rely on determining a distance to the workpiece and/or may utilize transform processing, etc.

The present disclosure includes systems and methods for capture a whole slide image of a sample. In exemplary embodiments, a camera is configured to capture a digital image of a sample. The system captures a bright field image of the sample, and captures a digital image of the sample illuminated from a first incident angle at a first wavelength and a second incident angle at a second wavelength. The system can determine whether the sample is defocused based on the transitional shift between a first wavelength channel and a second wavelength channel of the captured digital image. The system can determine the defocus distance based on the transitional shift and autofocus using the defocus distance such the bright field image is in focus.

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.

A fluorescence observation apparatus according to an embodiment of the present technology includes a stage, an excitation section, and a spectroscopic imaging section. The stage is capable of supporting a fluorescently stained pathological specimen. The excitation section irradiates the pathological specimen on the stage with a plurality of line illuminations of different wavelengths, the plurality of line illuminations being a plurality of line illuminations situated on different axes and parallel to a certain-axis direction. The spectroscopic imaging section includes at least one imaging device capable of separately receiving pieces of fluorescence respectively excited with the plurality of line illuminations.

A microscopic imaging method for creating a microscopic sample image (1A) of a sample (1) comprises the steps of arranging the sample (1) on a sampling crystal (10); irradiating the sample (1) with excitation laser pulses (2, 3) and generating sample response pulses (4) with a sample response field as a result of an interaction of the excitation laser pulses (2, 3) with the sample (1); irradiating the sampling crystal (10) with probe laser pulses (5) being temporally synchronized with the excitation laser pulses (2, 3) and spatially overlapped with the sample response pulses (4) in the sampling crystal (10), wherein the probe laser pulses (5) have a shorter wavelength than the excitation laser pulses (2, 3); detecting the sample response field by electric-field sampling with the sampling crystal (10), using the sample response pulses (4) and the probe laser pulses (5); and calculating the sample image (1A) based on the detected sample response field, wherein the excitation laser pulses (2, 3) have a wavelength in a range from mid-infrared to visible light and the sample response pulses (4) are created by a coherent interaction process induced in the sample (1) and with a fixed phase relationship relative to the excitation laser pulses (2, 3), the sampling crystal (10) is a non-centrosymmetric crystal, the irradiating step is repeated at multiple sample points (1A), wherein at each sample point (1A) the irradiating steps are successively repeated with multiple temporal probe delays of the probe laser pulses (5) relative to the excitation laser pulses (2, 3), at each probe delay, a sum or difference frequency pulse (6) of a sample response pulse (4) and a probe laser pulse (5) is generated, and at each probe delay, a spectral interference pulse (7) is created by a spectral interference of the sum or difference frequency pulse (6) and the current probe laser pulse, the detecting step includes sensing a polarization state of the spectral interference pulse (7) by an ellipsometer device (40) at each probe delay, wherein the local sample response field at the sample point (1A) is derived from the polarization states sensed at all probe delays, and the sample image (1A) is calculated based on the sample response field detected at the sample points (1A). Furthermore, a microscopic imaging apparatus is described.

The invention concerns a device for the holographic and hyperspectral measurement and analysis (2) of a sample (3), comprising; —an acquisition means (2) for acquiring a diffracted image (11) of an image of the sample (3); and interference patterns (12) of a reference light signal (R) and the light signal (O) having passed through the sample (3) to be measured and analysed; and—a means for illuminating the sample (3) focused on the sample (3); and—a means for reconstructing and analysing (1) the hyperspectral holographic image comprising a deep convolutional neural network generating an image for analysis and detection of particularities in the sample.

The multi-photon microscope comprises a repetition rate tuner that lowers an optical pulse train emitted from a pulsed laser to a repetition rate for time-gated detection, a scanner that scans the optical pulse train transmitted from the repetition rate tuner in x-axis and y-axis directions, an objective lens that irradiates an optical signal scanned by the scanner to the sample and acquires a fluorescence signal emitted from the excited fluorescent material, a photodetector that photoelectrically converts the fluorescence signal acquired by the objective lens, an amplifier that converts a current signal output from the photodetector into a voltage signal, a digitizer that samples the voltage signal output from the amplifier, and a computing device that separates sampling data output from the digitizer with a detection window set in time domain, and generates an image based on the sampling data separated by the detection window.

A laser scanning microscope and method for adjusting a laser scanning microscope. The microscope has an optical system which has a light guiding fiber between the first light source and the third beam deflection unit and has no light guiding fibers between the second light source and the third beam deflection unit. In this way, the second light source can be used as an adjustment reference for the first and second beam deflection units. The adjustment can be implemented using test images recorded by means of the third and fourth beam deflection units; additional sensors or internal calibration samples are not required.