Molecular Devices Corporation

Life Technologies Corporation

Matthew Chan

David Cox

Jon A Hoshizaki

Patrick D Kinney

Avrum Isaac Cohen

Albert L Carrillo

Howard Greg King

Marc Haberstroh

Steven Luke

Maryam Shariati

Joe Lee

Nia W Fong

Robert Malcolm Watson, Jr

Avrum L Cohen

A method and system for autofocusing an objective lens in a microscope system are disclosed. A decentered aperture is disposed in an optical path between an objective lens and an image plane of an image capturing device and a plurality of reference images are captured. Each reference image is captured when the objective lens is positioned at a corresponding z-position of a plurality of z-positions along an axis of travel of the objective lens and the optical path is at least partially occluded by the decentered aperture. At least one reference image of the plurality of the reference images is associated with a best focus position. The plurality of reference images are analyzed to develop a plurality of pattern locations, wherein each pattern location represents a position of a pattern formed on the image plane when a corresponding reference image was captured. The objective lens is positioned in accordance with the best focus position and the plurality of pattern locations.

Self-calibrating and directional focusing systems and methods for infinity corrected microscopes

Microscope imaging system and method for bolus formation and detection. In an exemplary method, immersion fluid may be delivered onto a front end of an objective of the imaging system to form one or more boluses. Each bolus may have a rounded outer surface intersected by an optical axis defined by the objective. At least a portion of each bolus may be illuminated. Light may be detected from each illuminated bolus to create detection data. A value representing a size of each bolus may be determined using the detection data. Delivery of the immersion fluid may be adjusted, if needed, based on the value(s) determined.

System and method for formation and detection of immersion fluid boluses on a microscope objective

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.

Imaging system and method using a projected reference to guide adjustment of a correction optic

Systems and methods for autofocusing an objective lens in a microscope system. A self-calibrating autofocus system includes a light source, a decentered aperture, and image-capturing device. The system may be connected to the microscope system so that the light source generates a light on an optical path that includes the objective lens and a plate having a reference surface proximal to a sample holding component. The image capturing device images a reflection of light from the reference surface as the objective lens moves to a series of z-positions. A reference calibration slope is generated for the objective lens by determining positions of the images taken at the z-positions of the objective lens. At least one image having a particular attribute corresponds to a best focus position. The objective lens is moved to predicted best focus or preferred offset from calibrated best focus using the reference calibration slope.

Method and system for trans-illumination imaging. In an exemplary method, a sample is irradiated in a well. The sample may include biological cells and a liquid or semi-solid medium that forms a meniscus. The step of irradiating may be performed with an array of light sources generating light of respective light beams that are incident on the meniscus at different orientations from one another. One or more images may be detected. A proportional contribution of light from two or more subsets of the light sources to the one or more images may be controlled by differential energization of the two or more subsets relative to one another to compensate for refraction by the meniscus and improve contrast.

Trans-illumination imaging with an array of light sources

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.

Imaging system and method with scattering to reduce source auto-fluorescence and improve uniformity

Imaging systems and methods using an ancillary image detector for sample location. An exemplary system may comprise a light source to irradiate a sample on an irradiation axis, a first image detector on an optical axis oblique to the irradiation axis, and a stage. The system also may comprise a second image detector disposed on an imaging axis, and a drive mechanism configured to move the stage and the imaging axis relative to one another. The system further may comprise a processor configured to (a) receive an image of the sample detected by the first image detector, (b) determine a physical location for a region of contrast produced by the sample within the image, and (c) send a signal to the drive mechanism based on the physical location, to dispose at least part of the sample in a field of view of the second image detector.

Imaging system with ancillary image detector for sample location

System of, and method for, trans-illumination imaging with use of interference fringes to enhance contrast and/or find focus. In an exemplary method, a coherence of light may be reduced upstream of a sample by scattering and mixing at least a portion of the light. The sample may be irradiated with the light of reduced coherence. An image of the sample may be detected, with the image being created with at least a portion of the light of reduced coherence that has passed through a plane defined by the sample. Image detection may be performed with the sample sufficiently defocused to form interference fringes in the image. The step of reducing a coherence of light may attenuate the interference fringes.

Trans-illumination imaging with use of interference fringes to enhance contrast and find focus

Microscope system for, and methods of, imaging a sample including biological cells. In an exemplary method, light transmitted through the sample may be detected for a first set of focal positions to collect a first stack of images. Values of a focus metric may be calculated for the first stack of images. A candidate focal position may be determined based on the values. Photoluminescence may be detected from the sample for a second set of focal positions to collect a second stack of images. The second set of focal positions may define a smaller range than the first set of focal positions. At least one focal position of the second set of focal positions may be based on the candidate focal position. In other words, the candidate focal position may serve as a guide for finding a suitable photoluminescence focal position.

Microscope system with transillumination-based autofocusing for photoluminescence imaging

Various embodiments of a sequencing system capable of rapidly imaging samples at multiple wavelengths are provided herein. In one embodiment, the system includes a fast-indexing filter wheel having a plurality of excitation and emission filters capable of being rapidly rotated into and out of communication with an excitation source (e.g., an arc lamp, a laser. For example, the filter wheel can be configured to index in an amount of time falling within a range of about 40 ms to about 60 ms, preferably 50 ms. The system can also be configured to account for vibrations resulting from the quick starts and stops of the fast-indexing filter wheel as well as vibrations resulting from other sources. Various methods of rapidly imaging a sample at multiple wavelengths are also provided herein.

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Palo Alto, CA, United States of America

Matthew Chan