An analyzer of a component in a sample fluid includes an optical source and an optical detector defining a beam path of a beam, wherein the optical source emits the beam and the optical detector measures the beam after partial absorption by the sample fluid, a fluid flow cell disposed on the beam path defining an interrogation region in the a fluid flow cell in which the optical beam interacts with the sample fluid and a reference fluid; and wherein the sample fluid and the reference fluid are in laminar flow, and a scanning system that scans the beam relative to the laminar flow within the fluid flow cell, wherein the scanning system scans the beam relative to both the sample fluid and the reference fluid.

The disclosed subject matter includes devices and systems for extending the imaging capability of swept, confocally aligned planar excitation (SCAPE) microscopes to in vivo applications. In embodiments, the SCAPE microscope can be implemented as an endoscopic or laparoscopic inspection instrument.

An imaging apparatus is provided to facilitate epifluorescent imaging of three (or more) color channels and to perform phase contrast and/or bright field imaging of samples without manual adjustment of the imaging apparatus. This allows for automated imaging, over extended periods of time, of a plurality of samples by a device located inside an incubator without disturbing the incubator environment to manually adjust the apparatus. Also provided are embodiments to facilitate user swapping of removable optical modules and/or transillumination modules to allow the imaging apparatus to be adapted to different combinations of assays and/or fluorescent indicators so as to increase the variety of experiments and/or fluorescent dyes that can be imaged using the imaging apparatus.

A method for obtaining image data of a subject, including; a first scanning step including a plurality of steps W, each step W being a step of determining one place existing in a one-dimensional, two-dimensional, or three-dimensional first space; and a second scanning step of scanning insides of second spaces including at least one of the places, wherein the second scanning step includes a step X of randomly determining a location of an observation point and a step Y of obtaining a piece of image data for the observation point, and, at a time point of end of scanning an inside of one of the second spaces, the second space has a first region including 50% of observation points and a second region existing outside the first region and including remaining 50% of the observation points, the second region being larger than the first region by at least 15%.

A single plane illumination microscope having an illumination optical system for illuminating a sample located on a sample carrier in a medium, and which is parallel to a planar reference surface. The sample is illuminated by a light sheet via an illumination light path. A detection optical system has a detection beam path. The optical axes of the illumination and detection optical systems each define an angle that is not equal to zero degrees along with the normal to the reference surface. A barrier layer system includes at least one layer of a given material having a given thickness and separates the medium from the illumination and detection optical systems. A base area of the barrier layer system is in contact with the region that is accessible for illumination and detection activities, said base area running parallel to the reference surface. At least one corrective element in the illumination beam path and/or the detection beam path allows those aberrations to be reduced which are created when light to be detected or light for illuminating the sample penetrates interfaces of the barrier layer system at an angle. The microscope has means, which are independent of the generation of the light sheet, for applying, via at least one manipulation beam path, light intensity to the sample in substantially point-shaped regions of the light sheet plane or in a given volume that at least temporarily encompasses the light sheet plane.

Methods and systems are provided that provide multiple simultaneous beams at differing angles used in scanning a light sheet for single plane illumination and applications thereof. The use of multiple beams reduces shadows that would otherwise stretch across the entire illumination sheet. The multiple beams may be created with a spatial light modulator as part of the illumination system.

An illumination arrangement for a microscope includes an illumination input configured to inject an illumination beam bundle and an illumination output configured to output at least two partial beam bundles generated from the illumination beam bundle. At least one diffractive optical element is configured to split the illumination beam bundle into the at least two partial beam bundles that propagate along partial beam paths, and is configured to effect a relative change in respective propagation directions of the at least two partial beam bundles with respect to one another, such that the at least two partial beam bundles output by the illumination arrangement are non-collinear with respect to one another at the illumination output.

Disclosed is an optical micro-spectrometry system including an optical microscope, a spectrometry system and an optical system adapted to direct an excitation light beam on the sample through the at least one microscope objective and to collect a Raman or PL light beam from a sample. The optical micro-spectrometry system includes an imaging system configured for acquiring a first image and a second image of the sample, by reflection or transmission of an illumination beam from a sample surface, the first image having a large field of view and the second image having a small field of view, a processing system configured for determining an area in the first image corresponding to the second image, a display system configured for displaying the first image, the second image, and a third image representing the area in overlay on the first image.

Provided herein is a macroscope comprising an objective apparatus comprising a multifocal widefield optics comprising a plurality of optical components configured to focus on a plurality of planes. Also provided herein are methods for analyzing a three-dimensional specimen, the method comprising obtaining, via a macroscope, synchronous multifocal optical images of a plurality of planes of the three-dimensional specimen, wherein the macroscope comprises an objective apparatus comprising a multifocal widefield optics comprising a plurality of optical components configured to focus on a plurality of planes. The three-dimensional specimen can be a biological specimen, such as brain.

Methods and apparatus for obtaining 3D imaging of tissue involve scanning a focused laser beam in xz planes to obtain a set of xz plane images spaced apart in a y direction. The xz plane images are processed to correct distortions and motions and combined to provide 3D image data. Surface flatting is optionally performed. Imaging may be performed using a femtosecond (fs) laser beam. Different components of light returning from the tissue may be detected and processed to yield plural co-registered images using different imaging modalities, for example, reflective confocal microscopy (RCM), two photon fluorescence (TPF) and second harmonic generation (SHG).