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.

Provided herein are systems and methods for simultaneous imaging and stimulation using a microscope system. The microscope system can have a relatively small size compared to an average microscope system. The microscope can comprise in part an imaging light source and a stimulation light source. Light from the imaging light source and the stimulation light source can be spectrally separated to reduce cross talk between the stimulation light and the imaging light.

System and method using a projected reference to guide adjustment of a correction optic. In an exemplary method, a reference may be projected onto an imaging detector by propagation of light generally along an optical axis that extends from the reference, through an objective, to a surface of a sample holder, and from the surface, back through the objective, to the imaging detector. The light may propagate through an off-axis aperture located upstream of the imaging detector and spaced from the optical axis. A plurality of images of the reference may be captured using the imaging detector, and with a correction optic at two or more different settings. A setting for the correction optic may be selected based on the plurality of images, and a sample may be imaged while the correction optic has the selected setting.

The present disclosure discloses a method and a system for imaging. The method for imaging objects using the system for imaging. The system for imaging comprises a lens. The objects comprise a first object, a second object and a third object located at different positions on a first preset track. The method for imaging comprises: allowing the lens and the first preset track to move relatively in a first predetermined relationship to acquire a clear image of the third object using the system for imaging without focusing, the first predetermined relationship is determined by a focal plane position of the first object and a focal plane position of the second object. The aforementioned method for imaging is high in imaging efficiency and is capable of fast focusing according to the first predetermined relationship even if focus tracking fails so that the blurring of a photographed image due to defocusing is avoided.

A microscope, including an x motor, a y motor, and a z axis focusing driving mechanism that drive a stage to move; at least one knob operatively coupled to the x motor or y motor or the z axis focusing driving mechanism selectively via a controller; and at least one button coupled to the controller in communication. The controller is configured to disconnect a normal operative coupling between the knob and the z axis focusing driving mechanism and establish an operative coupling between the knob and the x motor or y motor when receiving a predetermined button signal, and restore the normal operative coupling between the knob and the z axis focusing driving mechanism and disconnect the operative coupling between the knob and the x motor or y motor when no longer receiving the predetermined button signal or when receiving a next button signal.

A method of investigating a sample using a wide-field multiphoton microscope with tunable excitation wavelength (λ) is disclosed. The sample is illuminated at some excitation wavelength. Images of the sample are acquired at wavelengths different from the excitation wavelength, e.g., at half the excitation wavelength for SHG microscopy, using an image sensor. Based on the obtained images, an autoalignment procedure is carried out for optimizing the position of the sample relative to the illumination and/or collection beam paths. An image or spectrum that has been obtained for the optimal relative position is stored. The procedure is repeated for multiple excitation wavelengths. The autoalignment procedure can comprise an autofocusing subprocedure to automatically optimize the position of the sample relative to a focal plane of at least one objective of the multiphoton microscope along a direction that is perpendicular to the focal plane, and an in-plane repositioning subprocedure for automatically optimizing the position of the sample within the focal plane.

Systems and methods configured for simultaneous imaging and stimulation using a microscope system. The microscope system can have a relatively small size compared to an average microscope system. The microscope can comprise in part an imaging light source and a stimulation light source. Light from the imaging light source and the stimulation light source can be spectrally separated to reduce cross talk between the stimulation light and the imaging light.

An automatic focus system for an optical microscope that facilitates faster focusing by using at least two offset focusing cameras. Each offset focusing camera can be positioned on a different side of an image forming conjugate plane so that their sharpness curves intersect at the image forming conjugate plane. Focus of a specimen can be adjusted by using sharpness values determined from images taken by the offset focusing cameras.

The present disclosure discloses a method and a system for imaging. The method for imaging objects using the system for imaging. The system for imaging comprises a lens. The objects comprise a first object, a second object and a third object located at different positions on a first preset track. The method for imaging comprises: allowing the lens and the first preset track to move relatively in a first predetermined relationship to acquire a clear image of the third object using the system for imaging without focusing, the first predetermined relationship is determined by a focal plane position of the first object and a focal plane position of the second object. The aforementioned method for imaging is high in imaging efficiency and is capable of fast focusing according to the first predetermined relationship even if focus tracking fails so that the blurring of a photographed image due to defocusing is avoided.

A microscope includes an optical imaging system with an adjustable corrector, a microscope drive, a position sensitive detector, an optical measuring system and a control unit. The optical measuring system configured to form first and second measuring light beams, direct the measuring light beams into an entrance pupil of the optical imaging system eccentrically with first and second distances to the optical axis thereof, receive first and second reflection light beams, and direct the reflection light beams onto the position sensitive detector. The control unit is configured to record positions of the reflection light beams on the position sensitive detector, and determine an aberration based on the recorded positions.