The present invention discloses a photon enhancement apparatus comprising a reflective component and 4f coherent imaging system, which increases a photon collection efficiency. The present invention also provides a microscope comprising said photon enhancement apparatus and methods of improving photon collection efficiency, signal-to-noise ratio, and/or optical resolution using the said photon enhancement apparatus.

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.

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.

A high content imaging system and a method of operating the high content imaging system are disclosed. A microscope has a first objective lens and a second objective lens, and an objective lens database has first and second transformation values associated with the first and the second objective lenses, respectively. A microscope controller operates the microscope with the first objective lens to develop first values of acquisition parameters. A configuration module automatically determines second values of the acquisition parameters using the first values of the acquisition parameters, first transformation values associated with the first objective lens, and second transformation values associated with the second objective lens. The microscope controller operates the microscope using the second objective lens and the second values of the acquisition parameters.

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.

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.

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.

Microscopic analysis of a sample includes a system using dark-field illumination. A mid-IR optical source generates a mid-infrared beam, which is directed onto the sample to induce a temperature change by absorption of the mid-infrared beam. A visible light source generates a light illuminating the sample on a substrate and creating a scattered field and a reflected field along a collection path of the system. A pupil mask is positioned along the collection path to block the reflected field while allowing the scattered field to pass therethrough. A camera is positioned at an end of the collection path to collect the scattered field and generate a dark-field image of the sample.