Harvard College

Other

Howard Hughes Medical Institute

Kenneth Jeffrey Hayworth

Jeff Lichtman

Richard Schalek

Narayanan Kasthuri

Juan Carlos Tapia

Amy Au Hayworth

Harald F Hess

Marshall Burns

Kim F McGinnis

C Shan Xu

David Reese Peale

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

Methods, apparatuses and systems for facilitating automated or semi-automated collection of tissue samples cut by a microtome. In one example, a collection apparatus may be moved back and forth between respective positions at which the collection apparatus is operatively coupled to a microtome so as to collect cut tissue samples, or routine access to the microtome is provided. Relatively easy movement and positioning of the collection apparatus is facilitated, while at the same time ensuring structural stability and appropriate alignment and/or isolation between the collection apparatus and the microtome. A fluid reservoir receives samples cut by the microtome, and the collection apparatus may collect samples via a conveyor-like substrate disposed near/in the reservoir. A linear movement of the substrate may be controlled based on a cutting rate of the microtome, and the fluid level in the reservoir may be automatically maintained to facilitate effective sample collection.

Methods, apparatuses and systems for collection of tissue sections

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

Methods, apparatus and systems for collecting thin tissue samples for imaging. Thin tissue sections may be cut from tissue samples using a microtome-quality knife. In one example, tissue samples are mounted to a substrate that is rotated such that thin tissue sections are acquired via lathing. Collection of thin tissue sections may be facilitated by a conveyor belt. Thin tissue sections may be mounted to a thin substrate (e.g., by adhering thin tissue sections to a thin substrate via a roller mechanism) that may be imaged, for example, by an electron beam (e.g., in an electron microscope). Thin tissue sections may be strengthened before cutting via a blockface thinfilm deposition technique and/or a blockface taping technique. An automated reel-to-reel imaging technique may be employed for collected/mounted tissue sections to facilitate random-access imaging of tissue sections and maintaining a comprehensive library including a large volume of samples.

Methods, apparatus and systems for production, collection, handling, and imaging of tissue sections

Methods, apparatus and systems for collecting thin tissue samples for imaging. Thin tissue sections may be cut from tissue samples using a microtome-quality knife. In one example, tissue samples are mounted to a substrate that is rotated such that thin tissue sections are acquired via lathing. Collection of thin tissue sections may be facilitated by a conveyor belt. Thin tissue sections may be mounted to a thin substrate (e.g., by adhering thin tissue sections to a thin substrate via a roller mechanism) that may be imaged, for example, by an electron beam (e.g., in an electron microscope). This tissue sections may be strengthened before cutting via a blockface thinfilm deposition technique and/or a blockface taping technique. An automated reel-to-reel imaging technique may be employed for collected/mounted tissue sections to facilitate random-access imaging of tissue sections and maintaining a comprehensive library including a large volume of samples.

An automatic taping lathe-microtome that produces a continuous ribbon of tissue by lathing an extremely thin strip off the surface of a cylindrical block containing a multitude of embedded tissue samples. Mechanisms are included for sandwiching this fragile ribbon of tissue between a pair of support tapes. Viewing holes are cut in the support tapes above and below each tissue slice such that the tapes act as slot grids allowing for direct viewing of each tissue slice in a transmission electron microscope (TEM). The resulting tissue-tape is placed on a spooling mechanism and fed into the beam of a TEM much like the film in a movie projector. This allows for random-access imaging of any section on the tape without requiring the TEM's vacuum be broken. This system is intended to give neuroscientists a tool to ultrastructure image large volumes of neural tissue and to trace multi-scale synaptic circuits.

Methods and apparatuses for the automated production, collection, handling, and imaging of large numbers of serial tissue sections

A method and apparatus automatically fabricates a three dimensional object from individual layers of fabrication material having a predetermined configuration. Successive layers are stacked in a predetermined sequence and affixed together to form the object. The fabrication material is carried on a substrate to a stacker. At the stacker the layers are stacked together, with each layer being successively affixed to the stack of previously affixed layers, and with the substrate removed from each layer after it is affixed.

Growing community of inventors

Ashburn, VA, United States of America

Kenneth Jeffrey Hayworth