A method and system for the imaging and localization of fluorescent markers such as fluorescent proteins or quantum dots within biological samples is disclosed. The use of recombinant genetics technology to insert “reporter” genes into many species is well established. In particular, green fluorescent proteins (GFPs) and their genetically-modified variants ranging from blue to yellow, are easily spliced into many genomes at the sites of genes of interest (GoIs), where the GFPs are expressed with no apparent effect on the functioning of the proteins of interest (PoIs) coded for by the GoIs. One goal of biologists is more precise localization of PoIs within cells. The invention is a method and system for enabling more rapid and precise PoI localization using charged particle beam-induced damage to GFPs. Multiple embodiments of systems for implementing the method are presented, along with an image processing method relatively immune to high statistical noise levels.

System and method for localization of large numbers of fluorescent markers in biological samples

A system and method for aligning prior patterning positions formed by a first SPM tip with a second SPM tip in combination with an SPM system includes identifying first location information that includes a location of the first SPM tip and a sample reference location on an SPM sample and storing the first location information in a storage area. After replacing the first SPM tip with the second SPM tip, second location information, which includes a location of the second SPM tip and the sample reference location on the SPM sample, is identified. Displacement is calculated between the location of the second SPM tip and the first SPM tip based on the first and second location information, and either the second SPM tip or a stage supporting the SPM sample is translated to align the second SPM tip with the location of the first SPM tip in accordance with the calculated displacement.

Method and apparatus for aligning patterns on a substrate

The invention relates to a composite structure of a sample carrier 20 and a sample holder 30 for use in a TEM, for example. The sample carrier is hereby separately embodied from the sample holder. Although such compositions are already known, the known compositions are very fragile constructions. The sample carrier according to the invention can be formed from a strip of metal, and is a simple and cheap element. Using resilient force, it clamps onto or into the sample holder. The portion of the sample holder to which the sample carrier couples also has a simple form. The sample carrier can couple to the sample holder in vacuum using a coupling tool.

Sample carrier and sample holder

The invention provides a method for the manufacture and transmissive irradiation of a sample (16), comprising the steps of: A Providing a particle-optical system having an internal low-pressure chamber and suitable for the generation of an electron beam (22) and an intersecting ion beam (23) in said chamber; B Providing a specimen within the chamber, carried by a manipulator (1); C Irradiating the specimen (30) with the ion beam (23) so as to cut a sample (16) from the specimen; D Relatively displacing the sample (16) thus cut to a sample holder (11) than can be manipulated; E Attaching the sample to the sample holder (11 ); F Using an electron beam (22) to perform transmissive irradiation of the sample (16) thus attached to the sample holder (11), characterized in that step F is performed in the low-pressure chamber of the particle-optical system according to step A

Method for the manufacture and transmissive irradiation of a sample, and particle-optical system

An improved method and apparatus for S/TEM sample preparation and analysis. Preferred embodiments of the present invention provide improved methods for TEM sample creation, especially for small geometry (<100 nm thick) TEM lamellae. A novel sample structure and a novel use of a milling pattern allow the creation of S/TEM samples as thin as 50 nm without significant bowing or warping. Preferred embodiments of the present invention provide methods to partially or fully automate TEM sample creation, to make the process of creating and analyzing TEM samples less labor intensive, and to increase throughput and reproducibility of TEM analysis.

Method for creating s/tem sample and sample structure

An optical microscope has an optical axis that coincides with a focused ion beam. The optical microscope can be used to locate target features on a specimen for subsequent operations by the ion beam, thereby eliminating the need for complex and potentially inaccurate registration procedures. The optical microscope can use infrared light so that features on a silicon flip chip are observable through the silicon from the backside. The ion beam can then machine the chip to expose the features for subsequent operations.

Method and apparatus for a coaxial optical microscope with focused ion beam

A sample manipulator that includes a shaft, a proximal end, a distal end that includes a sample holder and a support arm having both a retracted position and a deployed position The support arm is configured to support a sample associated with the sample holder when the support arm is in the deployed position

Sample manipulation system

A multi-positional valve is used to control the destination of gas flows from multiple gas sources. In one valve position the gases flow to an isolated vacuum system where the flow rate and mixture can be adjusted prior to introduction into a sample vacuum chamber. In another valve position the pre-mixed gases flow from the isolated vacuum chamber and through a needle into the sample vacuum chamber.

Multiple Gas Injection System

An end effector for supporting a microsample during instrumental analysis, including a generally planar body and a cantilever tip configured to be associated with the microsample.

Jorgen Rasmussen

Papers

Method and apparatus for a coaxial optical microscope with focused ion beam

Sample manipulation system

Multiple Gas Injection System

End effector for supporting a microsample