The present disclosure relates a low magnification dark-field microscope system and method for producing a dark-field image. The method includes transferring a biological specimen to a surface of a sample plate, and pre-treating the biological specimen using one or more pre-treatment steps selected from (1) heating the biological specimen using a heating device; (2) applying ultrasound energy using an ultrasound transducer and ultrasound generator; and (3) doping the biological specimen with a metallic nanoparticle. Following pre-treatment, the method includes imaging a region of interest the biological specimen on the sample plate using a dark-field microscope to generate a dark-field image of the biological specimen.

An illuminator includes a light source and an illumination optical system for causing light emitted from the light source to be incident on a sample surface where an imaging object is present. The illumination optical system has an optical axis coaxial with that of an imaging optical system. An image of the light source is formed between the illumination optical system and the imaging optical system. A holder arranges the sample surface between the light source image and the imaging optical system.

Volumetric imaging with selective volume illumination (SVI) using light field detection is provided using various systems and techniques. A volumetric imaging apparatus includes a light source configured to emit an illumination light that propagates via an illumination light path to illuminate a three-dimensional (3D) sample; and an optical system arranged with respect to the light source to receive a light field, which comes from the illuminated 3D sample. The light field propagates via a detection light path, and the light source, the optical system, or both, are configurable to perform SVI, which selects a volume of a 3D-confined illumination of the 3D sample based on the 3D sample to be illuminated and a light field detection (LFD) process to be applied. Further, the volume of the 3D-confined illumination is a selected 3D volume of the 3D sample to be particularly excited by the 3D-confined illumination for imaging.

An observation apparatus, an observation method, and an observation program capable of capturing an image by appropriately adjusting a focus regardless of a size of an effective range in which a distance to a cultivation container can be measured within a field of view are provided. An observation apparatus includes an imaging unit 37 that images an observation target in a field of view smaller than an accommodation part 22 at a series of imaging positions and acquires a series of partial images, a measurement unit 38 that measures a distance from the imaging unit 37 to the accommodation part 22, a storage unit 44 that stores shape information of a container 20 and a series of imaging position information, a calculation unit 45 that calculates effective range information indicating an effective range in which the measurement unit 38 is capable of performing measurement before imaging within a field of view of the imaging unit 37 at the imaging positions, based on the shape information and the imaging position information, and a control unit 40 that controls a focus of the imaging unit 37 using a measurement result measured by the measurement unit 38 in the effective range and a measurement result of the measurement unit 38 in a field of view adjacent to the field of view including the effective range in a case where the effective range is smaller than or equal to a threshold value.

There are provided a microscopic mirror imaging device, a system and a method for calibrating a posture of a microneedle. The microscopic mirror imaging device includes a motion actuator, a mirror image former support and a mirror image former. The motion actuator is fixedly mounted on a microscope stage. One end of the mirror image former support is connected to the motion actuator. The mirror image former includes a plane mirror mounted on the other end of the mirror image former support. An angle formed between a mirror surface of the plane mirror and a horizontal plane of the microscope stage is equal to 45°, and an angle formed between the mirror surface of the plane mirror and a coronal plane of the microscope stage is equal to 45°.

The present disclosure provides automated robotic microscopy systems that facilitate high throughput and high content analysis of biological samples, such as living cells and/or tissues. In certain aspects, the systems are configured to reduce user intervention relative to existing technologies, and allow for precise return to and re-imaging of the same field (e.g., the same cell) that has been previously imaged. This capability enables experiments and testing of hypotheses that deal with causality over time with greater precision and throughput than conventional microscopy methods.

In various embodiments a module for generating an interference pattern for producing a digital holographic image is provided. The module comprises an adaptive lens arrangement configured to receive, from a microscope, an object wave of an intermediate image of a sample to be examined, and to generate an adapted object wave of the intermediate image of the sample by reducing a curvature of the object wave of the intermediate image; a reference input interface configured to receive an optical fiber delivering a reference wave from the coherent light source to the module and an interference arrangement configured to generate an interference pattern to be received by an imaging sensor arrangement, wherein the interference pattern is based on the adapted object wave and the reference wave from a coherent light source; wherein a position of the reference input interface of the module is configured to be adjustable with respect to at least two directions (x-y), wherein at least one of the adjustable directions is in parallel to a propagation direction of the reference wave leaving the optical fiber.

An imaging microscope (12) for generating an image of a sample (10) comprises a beam source (14) that emits a temporally coherent illumination beam (20), the illumination beam (20) including a plurality of rays that are directed at the sample (10); an image sensor (18) that converts an optical image into an array of electronic signals; and an imaging lens assembly (16) that receives rays from the beam source (14) that are transmitted through the sample (10) and forms an image on the image sensor (18). The imaging lens assembly (16) can further receive rays from the beam source (14) that are reflected off of the sample (10) and form a second image on the image sensor (18). The imaging lens assembly (16) receives the rays from the sample (10) and forms the image on the image sensor (18) without splitting and recombining the rays.

Certain disclosed embodiments concern an integrated imaging system that combined light-sheet microscopy, which enables considerable speed and phototoxicity gains, with quantitative-phase imaging. A method for using such imaging systems also is disclosed. In an exemplary embodiment, an integrated imaging system was used for multivariate investigation of live-cells in microfluidics.

A method of generating three-dimensional (3D) shape information of an object to be measured from an image including intensity information of an object hologram generated by interference between a reference light reflected from an optical mirror and an object light affected by the object includes checking at least one frequency component included in the image and extracting real image components corresponding to a real image from the frequency component. The method also includes generating a correction light and a real image hologram based on the real image components, generating an intermediate hologram based on the correction light, and generating curvature aberration correction information from the intermediate hologram. The method further includes generating a correction hologram based on the curvature aberration correction information and generating the 3D shape information of the object from the correction hologram.