A method for measuring distorted illumination patterns and correcting image artifacts in structured illumination microscopy. The method includes the steps of generating an illumination pattern by interfering multiple beams, modulating a scanning speed or an intensity of a scanning laser, or projecting a mask onto an object; taking multiple exposures of the object with the illumination pattern shifting in phase; and applying Fourier transform to the multiple exposures to produce multiple raw images. Thereafter, the multiple raw images are used to form and then solve a linear equation set to obtain multiple portions of a Fourier space image of the object. A circular 2-D low pass filter and a Fourier Transform are then applied to the portions. A pattern distortion phase map is calculated and then corrected by making a coefficient matrix of the linear equation set varying in phase, which is solved in the spatial domain.

A microscope for imaging a sample is disclosed that may include at least one illumination objective arranged to eject an illumination light beam along an illumination path to illuminate the sample; an imaging objective arranged to receive detection light including at least a portion of the light ejected from the sample, wherein the detection light is propagated along a detection axis and the imaging objective has an imaging focal plane; an adjustment arrangement to linearly displace the illumination light beam and the imaging focal plane relative to each other along the detection axis; a sample holder arranged to receive a sample and having a portion which is transparent to the illumination light beam and to the detection light; and a holder support arranged to receive the sample holder and displace the sample holder relative to the imaging objective such that the imaging focal plane is positioned inside the sample holder.

A method for illuminating samples in microscopic imaging methods, wherein a number m of different wavelengths λ.sub.i, with m>I and i=I, . . . , m, is selected for the illumination. For each of the wavelengths λ.sub.i a target phase function Δφ.sub.i(x, y, λ.sub.i) is predefined, wherein x and y denote spatial coordinates in a plane perpendicular to an optical axis z and each target phase function Δφ.sub.i(x, y, λ.sub.i) is effective only for the corresponding wavelength λ.sub.i. The target phase functions Δφ.sub.i are predefined depending on the structure of the sample and/or the beam shape and/or illumination light structure to be impressed on the light used for illumination. A total phase mask is then produced which realises all target phase functions Δφ.sub.i(x, y, λ.sub.i). This total phase mask is then illuminated simultaneously or successively with coherent light of wavelengths λ.sub.i such that the predefined structure of the illumination light is generated in the region of the sample.

Described herein are methods and systems for the optical imaging of a physical specimen of interest that is in contact with, or in close proximity to, the backplane of a high refractive index solid-immersion lens (SIL), wherein the specimen comprises features of interest that act as a local high-refractive index regions. The SIL lens preferably comprises fiducial markers.

A laser scanning microscope includes a light source configured to emit an illumination light beam. The illumination light beam has a transverse light intensity profile comprising an intensity minimum. The laser scanning microscope further includes a scanning device configured to scan the illumination light beam along a closed trajectory in a target area of a specimen, and a detector configured to detect fluorescence light emitted by a fluorophore within the target area of the specimen. The fluorophore is excited by the illumination light beam. The laser scanning microscope further includes a processor configured to determine an intensity distribution of the fluorescence light as a function of time and to determine a position of the fluorophore within the target area based on the intensity distribution of the fluorescence light.

A system, including an optical imaging assembly configured to image a sample at an object plane to an image plane; an image sensor arranged at the image plane and configured to capture images of the sample for a field of view of the system; a light source configured to emit light having a wavelength, λ; a spatial light modulator (SLM) arranged to receive the light emitted from the light source and to provide a spatially modulated light pattern; one or more optical elements arranged to receive the spatially modulated light pattern from the SLM and to direct the spatially modulated light pattern to the image plane; and an electronic controller in communication with the image sensor and the spatial light modulator, the electronic controller being programmed to identify one or more targets in the field of view of the optical imaging assembly and to control the spatial light modulator to selectively direct light from the light source to the one or more targets identified by the electronic controller.

There is provided a measurement apparatus including a control unit configured to control an on/off state of illumination that does not contribute to acquisition of measurement data on the basis of an acquisition time period of the measurement data.

A light sheet fluorescence microscope includes a light source configured to emit excitation light suitable for inducing fluorescent light emitted from a specimen, a detector configured to detect the fluorescent light from the specimen, and an optical system configured to illuminate the specimen with a light sheet formed from the excitation light, and to guide the fluorescent light from the illuminated specimen to the detector. The optical system includes an objective facing the specimen, the objective being configured to collect the fluorescent light emitted from the specimen. The light source is further configured to emit manipulation light suitable for photomanipulating the specimen. The optical system is further configured to direct the manipulation light through a spatially limited sub-area of an entrance pupil of the objective onto the specimen along a light propagation direction which is different from a light propagation direction of the light sheet.

Methods and systems for generating non-diffracting light sheets for multicolor fluorescence microscopy are disclosed. A method for generating a non-diffracting light patterned Bessel sheet comprises transmitting an input light beam through a Fourier transform lens the input light beam has a spatial intensity pattern at a first plane, and a Fourier plane is formed after the Fourier transform lens to obtain a first light beam; transmitting the first light beam through an annulus mask to obtain a second light beam; and transmitting the second light beam through an excitation objective lens to form a non-diffracting patterned light sheet. A method for generating a non-diffracting light line Bessel sheet comprises transmitting an input light beam at a first lane through an annulus mask to obtain a first light beam; and transmitting the first light beam through an excitation objective lens to form a non-diffracting Bessel light sheet.

A sample observation device includes: an emission optical system that emits planar light to a sample on an XZ plane; a scanning unit that scans the sample in a Y-axis direction so as to pass through an emission surface of the planar light; an imaging optical system that has an observation axis inclined with respect to the emission surface and forms an image of observation light generated in the sample; an image acquisition unit that acquires a plurality of pieces of XZ image data corresponding to an optical image of the observation light; and an image generation unit 8 that generates XY image data based on the plurality of pieces of XZ image data. The image generation unit extracts an analysis region of the plurality of pieces of XZ image data acquired in the Y-axis direction, integrates brightness values of at least the analysis region in a Z-axis direction to generate X image data, and combines the X image data in the Y-axis direction to generate the XY image data.