A device and a method for imaging a sample arranged in an object plane. The device includes an optical relay system that images an area of the sample from the object plane into an intermediate image plane. The device may also include an optical imaging system with an objective having an optical axis that lies perpendicularly on the intermediate image plan, and which is focused on the intermediate image plane, with the result that the object plane can be imaged undistorted onto a detector. The device also can include an illumination apparatus for illuminating the sample with a light sheet, wherein the light sheet lies essentially in the object plane and defines an illumination direction, and wherein the normal of the object plane defines a detection direction.

An image acquisition device includes a light source, a spatial light modulator having a plurality of pixels two-dimensionally arranged and fro modulating a phase of excitation light output from the light source for each of the plurality of pixels, a first objective lens, a second objective lens, a photodetector, and a control unit for controlling an amount of phase modulation for each of the plurality of pixels in accordance with a two-dimensional phase pattern corresponding to the plurality of pixels. The phase pattern is generated based on a predetermined basic phase pattern. The basic phase pattern includes a first region in which the phase value continuously increases in a predetermined direction and a second region in which the phase value continuously decreases in the direction and facing the first region in the direction.

Implementing swept, confocally aligned planar excitation (SCAPE) imaging with asymmetric magnification in the detection arm provides a number of significant advantages. In some preferred embodiments, the asymmetric magnification is achieved using cylindrical lenses in the detection arm that are oriented to increase the magnification of the intermediate image in the width direction but not in the depth direction. SCAPE imaging may also be improved by using an SLM to modify a characteristic of the sheet of excitation light that is projected into the sample. Additional embodiments include a customized version of SCAPE that is optimized for imaging the retina at the back of an eyeball in living subjects.

Arrangements and methods are provided for obtaining informationassociated with an anatomical sample. For example, at least one first electro-magnetic radiation can be provided to the anatomical sample so as to generate at least one acoustic wave in the anatomical sample. At least one second electro-magnetic radiation can be produced based on the acoustic wave. At least one portion of at least one second electro-magnetic radiation can be provided so as to determine information associated with at least one portion of the anatomical sample. In addition, the information based on data associated with the second electro-magnetic radiation can be analyzed. The first electro-magnetic radiation may include at least one first magnitude and at least one first frequency. The second electro-magnetic radiation can include at least one second magnitude and at least one second frequency. The data may relate to a first difference between the first and second magnitudes and/or a second difference between the first and second frequencies. The second difference may be approximately between 100 GHz and 100 GHz, excluding zero.

A system having a macroscopic imager, a microscopic imager, and a stage for moving a substrate supporting ex-vivo tissue with respect to each of the imagers to enable the macroscopic imager to capture macroscopic images, and the microscopic imager to capture optically formed sectional microscopic images on or within the tissue, when presented to the tissue, via the optically transparent material of the substrate. A computer system controls movement of the stage, and receives the macroscopic and microscopic images. A display is provided for displaying the macroscopic and microscopic images when received by the computer system. The tissue is verified as being in an orientation at least substantially flush against the upper surface of the substrate by being in focus in displayed macroscopic images prior to imaging by the microscopic imager, and if needed, any portion of the tissue unfocused is manually positioned until desired tissue orientation is achieved.

Optical sensing techniques and devices based on detection of fluorescent emissions at different optical wavelengths by nonlinear optical absorption of different excitation beams at different excitation wavelengths that interact with fluorescently-labeled structures within the sample to cause nonlinear optical absorption of two or more photons at each excitation wavelength. The fluorescent light at different fluorescent emission wavelengths by nonlinear optical absorption of excitation light at a particular excitation wavelength is spectrally separated into different optical channel output beams along different optical channel optical paths at different designated fluorescent imaging wavelength bands and the fluorescent light at different fluorescent imaging wavelengths within each designated fluorescent imaging wavelength is detected. This two-stage spectral separation in obtaining fluorescent images at different fluorescent imaging wavelengths in different fluorescent imaging wavelength bands enables highly sensitive hyperspectral imaging based on two-photo or multi-photon nonlinear absorption.

A microscope having three-dimensional imaging capability and a three-dimensional microscopic imaging method are provided, the microscope including: at least one excitation device configured to generate a detectable contrast in a detection target region of a sample which is to be detected, in an excitation principal axis direction; at least one detection device, configured to detect the contrast as generated from the detection target region of the sample in a detection principal axis; and at least one movement mechanism, configured to generate a relative movement of the sample relative to the excitation device and the detection device; the relative movement is in a direction neither parallel to nor perpendicular to the excitation principal axis direction or the detection principal axis direction.

A functionally integrated laser scanning microscope for scanning a sample with laser illumination, selectably in a confocal, line or wide-field operating mode, comprising a laser light source, an illumination and detection beam path, a detection device and at least one objective, wherein the illumination and detection beam path has optical means for the configuration of the laser illumination, at least one scanner for scanning the sample with the laser illumination, and a beam splitter for separating illumination and detection light, and controllable optical elements for changing the beam guiding depending on the operating mode selected in each case.

A system comprising: a dielectric configured to: a) create a bi-periodic interference pattern of two standing sinusoidal waves on illumination by two pairs of counter-propagating light beams at different incident angles, wherein the incident angles are selected in accordance with the index of refraction of the dielectric to i) to determine the spatial frequency of each counter-propagating light wave pair, and ii) cause total internal reflection, and b) generate, from the bi-periodic interference pattern, an evanescent bi-periodic standing sinusoidal wave; a light source configured to illuminate the dielectric with the two pairs of counter-propagating sinusoidal light waves at the selected incident angles and thereby illuminate a fluorescing object positioned at the surface of the dielectric with the generated bi-periodic evanescent standing sinusoidal wave; and one or more delay lines configured to independently modify the initial phase of each counter-propagating light wave pair.

Arrangements and methods are provided for obtaining information associated with an anatomical sample. For example, at least one first electro-magnetic radiation can be provided to the anatomical sample so as to generate at least one acoustic wave in the anatomical sample. At least one second electro-magnetic radiation can be produced based on the acoustic wave. At least one portion of at least one second electro-magnetic radiation can be provided so as to determine information associated with at least one portion of the anatomical sample. In addition, the information based on data associated with the second electro-magnetic radiation can be analyzed. The first electro-magnetic radiation may include at least one first magnitude and at least one first frequency. The second electro-magnetic radiation can include at least one second magnitude and at least one second frequency. The data may relate to a first difference between the first and second magnitudes and/or a second difference between the first and second frequencies. The second difference may be approximately between 100 GHz and 100 GHz, excluding zero.