Methods, devices, systems, and apparatuses are provided for the image analysis of measurement of biological samples.

Methods, systems and devices for automatically focusing a microscope on a specimen and collecting a focused image of the specimen are provided. Aspects of the methods include detecting the presence of a substrate in a microscope, determining whether the substrate is in a correct orientation for imaging, focusing the microscope on a specimen that is placed on the substrate, and collecting one or more images of the specimen. Systems and devices for carrying out the subject methods are also provided.

Methods, devices, apparatus, and systems are provided for image analysis. Methods of image analysis may include observation, measurement, and analysis of images of biological and other samples; devices, apparatus, and systems provided herein are useful for observation, measurement, and analysis of images of such samples. The methods, devices, apparatus, and systems disclosed herein provide advantages over other methods, devices, apparatus, and systems.

A microscope includes a movable stage supporting wells arranged in an array, a first imaging unit having a low-magnification objective lens, a second imaging unit having a high-magnification objective lens, a computer determining a representative position of a spheroid based on imaging data of the spheroid acquired by the first imaging unit, and a controller causing respective imaging units to sequentially acquire imaging data of the spheroid in each of the wells. The controller causes the first imaging unit to acquire imaging data for the spheroid in one of the wells, and then causes the stage to adjust the representative position to the optical axis of the high-magnification objective lens, and further causes the second imaging unit to acquire imaging data while causing the first imaging unit to acquire imaging data of the spheroid in another of the wells in synchronization with acquisition by the second imaging unit.

Imaging systems and methods with scattering to reduce source auto-fluorescence and improve uniformity. In some embodiments, the system may include a plurality of trans-illumination light sources configured to irradiate an examination region with different colors of trans-illumination light, while a same diffuser is present in each optical path from the trans-illumination light sources to the examination region. The system also may comprise an excitation light source configured to irradiate the examination region with excitation light. The system may be configured to irradiate the examination region with each of the trans-illumination light sources and, optionally, with the excitation light source, without moving parts in any of the optical paths from the trans-illumination light sources. The system further may comprise an image detector configured to detect grayscale images of the examination region, and a processor configured to create a color trans-illumination image from grayscale images.

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.

Provided is a microscope system including: a stage on which a multi-dyed specimen is mounted; an objective lens for collecting light from the specimen mounted on the stage; a Z-axis movement section for relatively moving the stage and the objective lens in a direction along the optical axis L of the objective lens; an XY-axis movement section for moving the stage in a direction orthogonal to the optical axis L; an image acquisition unit for acquiring a color image by capturing the light collected by the objective lens; and a depth-extension processing unit for generating a depth-extended image by performing depth extension processing dye by dye on the basis of a plurality of the color images that are acquired by the image acquisition unit at different positions of the stage relative to the objective lens set with the Z-axis movement section.

A spectral imaging device (12) includes an image sensor (28), a tunable light source (14), an optical assembly (17), and a control system (30). The optical assembly (17) includes a first refractive element (24A) and a second refractive element (24B) that are spaced apart from one another by a first separation distance. The refractive elements (24A) (24B) have an element optical thickness and a Fourier space component of the optical frequency dependent transmittance function. Further, the element optical thickness of each refractive element (24A) (24B) and the first separation distance are set such that the Fourier space components of the optical frequency dependent transmittance function of each refractive element (24A) (24B) fall outside a Fourier space measurement passband.

An observation device includes an illuminating optical system that emits illuminating light obliquely upward from below a sample; and an objective optical system that images transmitted light, which is the illuminating light emitted from the illuminating optical system, reflected above the sample, and passed through the sample, the objective optical system imaging the transmitted light below the sample and via a different path from that of the illuminating optical system. The illuminating optical system includes a light source, a mask that restricts light, which is emitted from the light source, to a particular emission region, and a collimating optical system that converts the light restricted by the mask into substantially parallel light, and the illuminating optical system is arranged so that when the region is projected onto a pupil plane of the objective optical system, a projected image of the region partially overlaps a peripheral portion of the pupil.

Systems and methods for three-dimensional fluorescence polarization excitation that generates maps of positions and orientation of fluorescent molecules in three or more dimensions are disclosed.