Abstract: A charged-particle microscope, comprising a vacuum chamber in which are provided: A specimen holder for holding a specimen in an irradiation position; A particle-optical column, for producing a charged particle beam and directing it so as to irradiate the specimen; A detector, for detecting a flux of radiation emanating from the specimen in response to irradiation by said beam, wherein: Said vacuum chamber comprises an in situ magnetron sputter deposition module, comprising a magnetron sputter source for producing a vapor stream of target material; A stage is configured to move a sample comprising at least part of said specimen between said irradiation position and a separate deposition position at said deposition module; Said deposition module is configured to deposit a layer of said target material onto said sample when held at said deposition position.

Abstract: A charged-particle microscope, comprising a vacuum chamber in which are provided: A specimen holder for holding a specimen in an irradiation position; A particle-optical column, for producing a charged particle beam and directing it so as to irradiate the specimen; A detector, for detecting a flux of radiation emanating from the specimen in response to irradiation by said beam, wherein: Said vacuum chamber comprises an in situ magnetron sputter deposition module, comprising a magnetron sputter source for producing a vapor stream of target material; A stage is configured to move a sample comprising at least part of said specimen between said irradiation position and a separate deposition position at said deposition module; Said deposition module is configured to deposit a layer of said target material onto said sample when held at said deposition position.

Abstract: The present invention relates to a method of studying a sample using an optical microscope, comprising providing the sample in a sample holder with means to maintain the sample at a temperature below 273 K; providing a microscope objective lens, in a thermally insulating jacket, having an extremal lens element proximal to the sample holder; bringing the lens into a focus position proximal to the sample, which separates the extremal lens element and sample by an intervening space, providing a transparent window in said intervening space, with a gap between the window and the extremal lens element; providing a flow of substantially dry gas in said gap; and tailoring the geometry and velocity of said flow so that, at least in said gap, the flow is non-laminar; and does not excite substantial acoustic vibration in a structure proximal the gap.

Abstract: A method for Transmission Electron Microscopy (TEM) sample creation. The use of a Scanning Electron Microscope (SEM)—Scanning Transmission Electron Microscope (STEM) detector in the dual-beam focused ion beam (FIB)/SEM allows a sample to be thinned using the FIB, while the STEM signal is used to monitor sample thickness. A preferred embodiment of the present invention can measure the thickness of or create TEM and STEM samples by using a precise endpoint detection method. Preferred embodiments also enable automatic endpointing during TEM lamella creation and provide users with direct feedback on sample thickness during manual thinning. Preferred embodiments of the present invention thus provide methods for endpointing sample thinning and methods to partially or fully automate endpointing.

Abstract: The invention relates to a method of welding a vitreous biological sample at a temperature below the glass transition temperature of approximately ?137° C. to a micromanipulator, also kept at a temperature below the glass transition temperature. Where prior art methods used IBID with, for example, propane, or a heated needle (heated resistively or by e/g/laser), the invention uses a vibrating needle to locally melt the sample. By stopping the vibration, the sample freezes to the micromanipulator. The heat capacity of the heated parts is small, and the amount of material that stays in a vitreous condition thus large.

Abstract: The invention relates to a method of welding a vitreous biological sample at a temperature below the glass transition temperature of approximately ?137° C. to a micromanipulator, also kept at a temperature below the glass transition temperature. Where prior art methods used IBID with, for example, propane, or a heated needle (heated resistively or by e/g/laser), the invention uses a vibrating needle to locally melt the sample. By stopping the vibration, the sample freezes to the micromanipulator. The heat capacity of the heated parts is small, and the amount of material that stays in a vitreous condition thus large.

Abstract: The invention relates to a method of forming a vitrified sample on a sample holder for inspection in an electron microscope. It is known to spray a solution on a grid and then immerse the grid in a cryogenic liquid, such as ethane or liquid nitrogen. The invention proposes to spray small droplets of the liquid on a cryogenic surface, such as a grid or a sample holder in vacuum. The liquid forms vitrified sample material when hitting the surface due to the low temperature of the grid or sample holder. A lamella may be excavated from the thus formed sample material, to be studied in a TEM, or the vitrified sample material may be directly observed in a SEM. In an embodiment the material may be sprayed on a cryogenic liquid, to be scooped from the liquid and placed on a grid.

Abstract: The present invention relates to a method of studying a sample using an optical microscope, comprising providing the sample in a sample holder with means to maintain the sample at a temperature below 273 K; providing a microscope objective lens, in a thermally insulating jacket, having an extremal lens element proximal to the sample holder; bringing the lens into a focus position proximal to the sample, which separates the extremal lens element and sample by an intervening space, providing a transparent window in said intervening space, with a gap between the window and the extremal lens element; providing a flow of substantially dry gas in said gap; and tailoring the geometry and velocity of said flow so that, at least in said gap, the flow is non-laminar; and does not excite substantial acoustic vibration in a structure proximal the gap.

Abstract: Described is a system and method for in situ sample preparation and imaging. The system includes a multi-axis stage 100 having a bulk stage 110 and a grid stage 150 with various degrees of freedom to allow for sample preparation. In some embodiments, a focused ion beam system is used to prepare a lamella on the bulk stage 110. The lamella can then be transferred to the grid stage 150 from the bulk stage 110 without needing to move the multi-axis stage 100 from the focused ion beam system.

Abstract: A method of forming a sample from a capillary with high-pressure frozen sample material comprises providing a high-pressure capillary with vitrified sample material at a temperature T1 below the glass transition temperature Tg, cutting the capillary, warming the capillary to a temperature T2 between temperature T1 and temperature Tg, cooling the capillary to a temperature T3 below temperature T2, as a result of which material is extruded from the capillary, and freeing a sample from the extruded sample material at a temperature below temperature Tg. Repeating this temperature cycle results in further extrusion of the sample material. The extruded material can then be sliced by, for example, ion beam milling.

Abstract: The invention relates to an assembly of a cooperating capillary and cap for containing an aqueous solution in an inner volume of the capillary. The assembly is used in a high pressure freezer in which the aqueous solution is frozen at a high pressure to form an amorphous frozen sample at a cryogenic temperature. The cap forms a closure at one end of the capillary, and the part of the cap that is in contact with the inner volume of the capillary has an indent; as a result of which the cap, after freezing the aqueous solution, can be removed from the capillary and a free standing pillar of frozen aqueous material extends from the capillary.

Abstract: A method of forming a sample from a capillary with high-pressure frozen sample material comprises providing a high-pressure capillary with vitrified sample material at a temperature T1 below the glass transition temperature Tg, cutting the capillary, warming the capillary to a temperature T2 between temperature T1 and temperature Tg, cooling the capillary to a temperature T3 below temperature T2, as a result of which material is extruded from the capillary, and freeing a sample from the extruded sample material at a temperature below temperature Tg. Repeating this temperature cycle results in further extrusion of the sample material. The extruded material can then be sliced by, for example, ion beam milling.

Abstract: A method for Transmission Electron Microscopy (TEM) sample creation. The use of a Scanning Electron Microscope (SEM)—Scanning Transmission Electron Microscope (STEM) detector in the dual-beam focused ion beam (FIB)/SEM allows a sample to be thinned using the FIB, while the STEM signal is used to monitor sample thickness. A preferred embodiment of the present invention can measure the thickness of or create TEM and STEM samples by using a precise endpoint detection method. Preferred embodiments also enable automatic endpointing during TEM lamella creation and provide users with direct feedback on sample thickness during manual thinning. Preferred embodiments of the present invention thus provide methods for endpointing sample thinning and methods to partially or fully automate endpointing.

Abstract: A method for Transmission Electron Microscopy (TEM) sample creation. The use of a Scanning Electron Microscope (SEM)—Scanning Transmission Electron Microscope (STEM) detector in the dual-beam focused ion beam (FIB)/SEM allows a sample to be thinned using the FIB, while the STEM signal is used to monitor sample thickness. A preferred embodiment of the present invention can measure the thickness of or create TEM samples by using a precise endpoint detection method. Preferred embodiments also enable automatic endpointing during TEM lamella creation and provide users with direct feedback on sample thickness during manual thinning. Preferred embodiments of the present invention thus provide methods for endpointing sample thinning and methods to partially or fully automate endpointing.

Abstract: An improved method for TEM sample creation. The use of a SEM-STEM detector in the dual-beam FIB/SEM allows a sample to be thinned using the FIB, while the STEM signal is used to monitor sample thickness. A preferred embodiment of the present invention can measure the thickness of or create S/TEM samples by using a precise endpoint detection method that is reproducible and suitable for automation. Preferred embodiments also enable automatic endpointing during TEM lamella creation and provide users with direct feedback on sample thickness during manual thinning. Preferred embodiments of the present invention thus provide improved methods for endpointing sample thinning and methods to partially or fully automate endpointing to increase throughput and reproducibility of TEM sample creation.

Abstract: The invention pertains to a method for separating a minute sample (1) from a work piece (2). Such a method is routinely used in the semiconductor industry to obtain samples from wafers to be inspected in a TEM. It occurred to the inventor that approximately 20% of the obtained samples could not be properly finished (thinned) due to a misalignment of specimen carrier (6) and sample. It turned out that this misalignment is caused by the specimen carrier contacting the sample prior to welding. By not contacting the sample while welding, but leaving a small gap between specimen carrier and sample, this misalignment is avoided. To avoid movement of the specimen carrier during welding, due to e.g. vibration, the specimen carrier can be landed on the wafer on a position (8) close to the sample.

Abstract: The invention pertains to a method for separating a minute sample (1) from a work piece (2). Such a method is routinely used in the semiconductor industry to obtain samples from wafers to be inspected in a TEM. It occurred to the inventor that approximately 20% of the obtained samples could not be properly finished (thinned) due to a misalignment of specimen carrier (6) and sample. It turned out that this misalignment is caused by the specimen carrier contacting the sample prior to welding. By not contacting the sample while welding, but leaving a small gap between specimen carrier and sample, this misalignment is avoided. To avoid movement of the specimen carrier during welding, due to e.g. vibration, the specimen carrier can be landed on the wafer on a position (8) close to the sample.