A method for acquiring an image, in which an image of an image acquiring region is acquired by radiating an ion beam to a sample having a conducting part with a linear edge on a dielectric substrate, includes performing an equal-width scan of the ion beam in a first direction that obliquely intersects the linear edge and sweep in a second direction intersecting the first direction. The ion beam is sequentially scanned in different patterns on different scan regions of parallelogram shape, each of which includes the image acquiring region. Secondary charged particles are detected to generate image data of all the scan regions, and image data of the scan regions are calculated to generate image data of the image acquiring region. The image data of the image acquiring region are synthesized to display the image data of the image acquiring region.

The present disclosure is directed to embodiments of microtome devices and methods of their use. In some embodiments, a microtome can be mounted on the built-in stage of a scanning electron microscope and used to perform serial block-face scanning electron microscopy. In some cases, a microtome installed in a scanning electron microscope can cut the sample at a location off the electron beam axis of the scanning electron microscope. In some cases, a microtome can include a capacitive sensor which can measure the location of a blade of the microtome, and the microtome can be computer-controlled by program implemented in MATLAB.

A sample is milled to expose the region of interest (ROI) within the sample, while being held by a sample stage in a microscopy system. The sample is milled based on the ROI location determined with sample images acquired with light beam irradiating from different axes. The sample images are acquired while the sample is held using the same sample stage for milling.

Specimen imaging systems and methods including a sample stage in a vacuum environment. The sample stage is configured to support a specimen, an electron beam generator configured to focus an electron beam on a first predetermined location on the specimen, a nanospray dispenser configured to dispense a nanospray onto a second predetermined location on the specimen, a mass spectrometer, and an extraction conduit configured to extract a plume of charged particles generated as a result of contact between the nanospray and the specimen and deliver the charged particles to the mass spectrometer. The systems and methods can create a topological and chemical map of the specimen by analyzing at least a portion of the specimen with a mass spectrometer to determine a chemical composition of the specimen at the second predetermined location and analyzing at least a portion of the specimen with the electron beam to determine a surface topology.

Disclosed are compositions and methods for the conductive fixation of organic material, including biological samples. The compositions and methods described herein can address the problems of charging and sample damage caused by electron beam-sample interactions within an electron microscope.

An imaging system that selectively alternates a first, non-destructive imaging mode and a second, destructive imaging mode to analyze a specimen so as to determine an atomic structure and composition of the specimen is provided. The field ionization mode can be used to acquire first images of ionized atoms of an imaging gas present in a chamber having the specimen disposed therein, and the field evaporation mode can be used to acquire second images of ionized specimen atoms evaporated from a surface of the specimen with the imaging gas remaining in the chamber. The first and second image data can be analyzed in real time, during the specimen analysis, and results can be used to dynamically adjust operating parameters of the imaging system.

Disclosed herein is a method for acquiring an image, in which an image reducing the influence of electrification of a substrate is easily acquired. The method, in which an image of an image acquiring region is acquired by radiating an ion beam to a sample having a conducting part with a linear edge on a dielectric substrate, includes: performing an equal-width scan caused by the ion beam in a first direction that obliquely intersects the edge and sweep in a second direction intersecting the first direction, and radiating the ion beam to a scan region of a parallelogram shape wider than the image acquiring region; detecting secondary charged particles to generate image data of the scan region; calculating the image data of the scan region to generate image data of the image acquiring region; and displaying the image data of the image acquiring region.

The present disclosure is directed to embodiments of microtome devices and methods of their use. In some embodiments, a microtome can be mounted on the built-in stage of a scanning electron microscope and used to perform serial block-face scanning electron microscopy. In some cases, a microtome installed in a scanning electron microscope can cut the sample at a location off the electron beam axis of the scanning electron microscope. In some cases, a microtome can include a capacitive sensor which can measure the location of a blade of the microtome, and the microtome can be computer-controlled by program implemented in MATLAB.

A method and a dual beam FIB/(S)TEM apparatus are provided for in-situ sample quality inspection in cryogenic focused ion beam milling. The method comprises the steps of: loading the sample into a sample holder of the dual beam FIB/(S)TEM apparatus, wherein the (S)TEM apparatus comprises an electron column and a detector, wherein the sample holder is arranged in between the electron column and the detector; obtaining an image of the electrons that have passed through the sample using the electron column to direct an electron beam towards the sample and using the detector to detect electrons passing through the sample; and using a scattering pattern in the image of the transmitted electrons to establish a measure for the thickness of the sample and to establish whether or not the image comprises a diffraction signal due to electron diffraction from ice crystals.

Disclosed are compositions and methods for the conductive fixation of organic material, including biological samples. The compositions and methods described herein can address the problems of charging and sample damage caused by electron beam-sample interactions within an electron microscope.