Kla-tencor Technologies Corporation

Hestzig LLC

Howard Hughes Medical Institute

Other

Kla Tencor Corporation

Hess, Harald F.

Betzig, Robert Eric

Harald F Hess

Eric Betzig

Marian Mankos

David Reese Peale

David Lewis Adler

Kirk Joseph Bertsche

David Aitan Soltz

Kenneth Jeffrey Hayworth

Patrick Rodney Lee

C Shan Xu

Shem-Tov Levi

Patrick T Lee

Marek Zywno

Gustavo A Pinto

Robert E Betzig

Vincenzo Lordi

Romain Sappey

Thomas Carr

Dan L Kilmer

Carlos A Duran

Alex Moraru

Robert Eric Betzig

Tom Carr

Jun Zhu

Teodor Zanetti

Franklin Tao

Matt Bellis

Gleb Shtengel

Kenneth J Hayworth

Imaging a sample that includes phototransformable optical labels ('PTOLs') with an optical system having a diffraction-limited resolution volume (DLRV), includes providing activation radiation to the PTOLs to activate a statistical subset of the PTOLs. A density of the PTOLs of the activated subset is less than an inverse of the DLRV. Excitation radiation is provided to the activated subset to excite activated PTOLs. Radiation emitted from the activated and excited PTOLs located at different focal planes of the optical system within the sample is detected with the optical system. The preceding steps are repeated one or more times, each time activating a different statistical subset of the plurality of PTOLs. Three-dimensional locations within the sample are determined, with a sub-diffraction-limited accuracy, of the activated and excited PTOLs based on the radiation emitted from the activated and excited PTOLs that is detected from the different focal planes of the optical system.

Optical microscopy with phototransformable optical labels

A microscopy system includes a gas cluster beam system configured for generating a beam of gas clusters directed toward a sample to irradiate a sample and mill away successive surface layers from the sample, a scanning electron microscope system configured for irradiating the successive surface layers of the sample with an electron beam and for imaging the successive surface layers of the sample in response to the irradiation of the surface layer, and a processor configured for generating a three dimensional image of the sample based on the imaging of the successive layers of the sample.

Volume scanning electron microscopy of serial thick tissue sections with gas cluster milling

A microscopy system for imaging a sample can include a scanning electron microscope system configured for imaging a surface layer of the sample and a focused ion beam system configured for generating an ion beam for milling the surface layer away from a sample after it has been imaged. A movable mechanical shutter can be configured to be moved automatically into a position between the sample and the scanning electron microscope system, so that when the electron beam is not imaging the sample the movable mechanical shutter is positioned between the sample and the scanning electron microscope system.

Enhanced FIB-SEM systems for large-volume 3D imaging

A microtome includes a blade located at an end of a trough that defines a cavity for holding a liquid; a sample block in which the at least one sample is suspended, the sample block is moveable relative to the blade such that when the sample block is passed across the blade a section is cut from the sample block; a plate that includes a support frame that defines an opening, and a transparent film extending across the opening, the transparent film being transparent to electrons, a grasper being configured to receive and retain the plate, wherein the grasper is moveable relative to the blade; and a pusher section that lacks the sample pusher section.

Sectioning volume samples

A method includes sequentially acquiring phase-shifted images of an optical source in a plurality of predetermined positions in the z plane to form a plurality of image frame sequences; determining from the plurality of image frame sequences at least one phase-related characteristic associated with each predetermined position of the optical source in the z plane; and storing the at least one phase-related characteristic and the predetermined positions of the optical source in the z plane.

Methods for 3-dimensional interferometric microscopy

In one embodiment, an apparatus comprises an optical system with multiple detectors and a processor. The optical system is configured to produce images of an optical source in a first dimension and a second dimension substantially orthogonal to the first dimension at each detector at a given time. Each image from the images is based on an interference of an emission from the optical source in a first direction and an emission from the optical source in a second direction different from the first direction. The processor is configured to calculate a position in a third dimension based on the images. The third dimension is substantially orthogonal to the first dimension and the second dimension.

Optical interfering apparatus

An apparatus includes a position-sensitive detector to detect intensities of radiation as a function of position on the detector, and an optical system, characterized by a diffraction-limited resolution volume, adapted for imaging light emitted from activated and excited phototransformable optical labels ('PTOLs') in a sample onto the position sensitive-detector. A first light source provides activation radiation to the sample to activate a subset of the PTOLs that are distributed in the sample with a density greater than an inverse of the diffraction-limited resolution volume of the optical system. A second light source provides excitation radiation to the sample to excite a portion of the PTOLs in the subset of the PTOLs. A controller controls one both of the activation radiation and the excitation radiation provided to the sample such that a density of PTOLs in the portion of the PTOLs is less than the inverse of the diffraction-limited resolution volume.

A statistically sparse subset of switchable optical sources in a sample is activated, and the activated switchable optical sources are excited such that optical beams are emitted from the activated switchable optical sources along at least two optical paths. A first wavefront modification in a first optical beam emitted from the activated switchable optical sources along a first optical path is introduced and a second wavefront modification in a second optical beam emitted from the activated switchable optical sources along a second optical path is introduced, the second wavefront modification being distinct from the first wavefront modification. The first and second optical beams are interfered with each other to produce a plurality of output beams, and three-dimensional position information of the optical sources is determined based on an intensity of each output beam from the plurality of output beams.

Three-dimensional interferometric microscopy

Systems and methods for 3-dimensional interferometric microscopy

One embodiment disclosed relates to a reflective electron patterning device. The device includes a pattern on a surface. There is an electron reflective portion of the pattern and an electron non-reflective portion of the pattern. Another embodiment disclosed relates to a method of reflecting a pattern of electrons. An electron beam is generated to be incident upon a surface. The pattern is formed on the surface. The incident electrons are reflected from a reflective portion of the pattern are prevented from being reflected from a non-reflective portion of the pattern.

Reflective electron patterning device and method of using same

Growing community of inventors

La Jolla, CA, United States of America

Harald F Hess