TWI433195B - Specimen supporting device for electron microscope and fabrication method thereof - Google Patents

Description

Sample carrying device for electron microscope and manufacturing method thereof

The present invention relates to a sample carrying device and a method of fabricating the same, and more particularly to a sample carrying device for an electron microscope and a method of fabricating the same.

Generally, the electron microscope needs to be observed in a vacuum environment. The sample needs to be exposed to a vacuum environment, and the sample to be observed needs to be dried before being observed. Therefore, samples containing liquid or volatile substances cannot be observed in an electron microscope.

When conducting experiments on physical, chemical, or biological samples that are sensitive to external environmental conditions, they must be placed in a control space with stable environmental variables to maintain constant temperature, humidity, pressure, concentration, pH, stress, and inertial force. Blocks interference caused by light source, electric field, magnetic field, sound, inertial force, vibration, to avoid unintended interference of the object under test, and changes in appearance or function. Generally, the environmental control device used, in addition to being able to isolate the external environment to protect the sample in the controlled state, does not affect the experimental observation. It is also necessary to allow physical quantities (visible light, laser, X-ray, electron beam, etc.) in the measurement mechanism to enter and exit the environmental control device. For example, when observing a liquid-containing test piece in an electron microscope, it is necessary to prevent the liquid or gas from being dissipated in the vacuum chamber, to avoid the interference of the vacuum required for electron beam imaging, and to keep the sample in a liquid state. The general practice is to confine the liquid to an environmental device and replace it with the original test piece and mount it in an electron microscope. A small hole is opened in the environmental device to give an electron beam through the path of the liquid and the environmental device, and the liquid or vaporized liquid caused by the small hole is restricted from leaking, and a so-called environmental cavity is formed inside the device.

The preparation method of the test piece of the conventional bioelectron microscope includes several basic steps: fixation in a formaldehyde solution, dehydration in alcohol or acetone, embedding in a resin, ultra-thin sectioning, heavy metal staining, etc., and observation by electron microscopy. Under normal electron microscope operation, it is impossible to observe samples containing water. Since the vacuum pressure in the column of the electron microscope is much smaller than the saturated vapor pressure of water, in order to maintain the stability of the electron beam, the wet sample is completely drained and dehydrated in the vacuum chamber. However, if the sample moisture is removed during the preparation of the test piece, it will cause unpredictable changes in the structure and morphology of the sample.

Since 1962, how to study the interaction between gas and sample and to observe wet samples in a transmission electron microscope has been proposed. To keep the aqueous sample moist during the observation process, the sample must be separated from the vacuum system to protect the sample from dehydration. Therefore, a specially designed cavity device is required to confine the liquid inside. There is a small hole in the upper and lower sides of the device for the electron beam to pass through. Depending on the design, the small hole may be a completely penetrating hole or may be left with gas. A small window of dense film, this is called the environmental cavity. There are two major development directions for environmental chamber devices: wet chambers with limited pores and wet chambers with thin films. The former adopts an open cavity design and uses very small pores as a passage of electron beams. During the operation of the microscope, the moisture of the sample may flow from the pores into the vacuum chamber, and the balance between the steam leakage rate of the pores and the gas supply rate of the wet chamber is adjusted. To maintain the presence of liquid in the wet room. The latter covers the pores with a film, and the liquid vapor does not leak into the vacuum chamber. The film needs to have sufficient electron penetration and sufficient mechanical strength and air tightness.

Around 1960, each team developed a film-limited environmental chamber, and the concepts were quite close. The environmental device is mounted on an electron microscope test strip stage, and is mainly composed of a gas tight gasket, a copper sheet having small holes, a sealing film, and a spacer. A sealing film is coated on the copper sheet to cover the small hole, and the two copper sheets are fixed by a gas-tight gasket to sandwich the spacer to form a closed environment device, and an external circulation line is provided to pass the gas into the device to control the pressure difference between the inside and the outside of the film. The thickness of the sample layer is affected by the height of the spacer. Generally, the copper piece with a diameter of 50 μm is 3.5 mm, the diameter of the middle hole is 300 μm, and the thickness of the film covering the small hole is at least 200 nm, and sufficient mechanical strength is required. . An excessively thick film will seriously affect the crossing of the electron beam, reducing brightness and resolution. The conventional technique of 1965 proposed the idea of adding a copper mesh to the film to improve the mechanical strength. The mesh was formed by a number of 60 micron diameter hollow holes, which greatly reduced the required film thickness and adopted a carbon film of 20 nm. As a sealing film, a high-resolution electron microscope image is first obtained in a film-limited environment device.

In 1968, the prior art developed a pore-limited sample device capable of high-temperature gas reaction, allowing electron microscopic observation of the sample in a gas atmosphere at 300 Torr and 1000 °C. The sample is placed on a mica annular sheet. The middle hollow portion of the mica sheet has a platinum heating wire and an electrode to form a sample platform that can be heated and monitored for temperature. The mica sample platform is housed in a metal gas reaction chamber and is sandwiched between two sheets of metal with tiny holes in between. There is a pipeline through the gas into the reaction chamber and fill the sample around, the excess gas is overflowed by two 50 micron tiny holes, and is pumped away by the microscope vacuum system. The brightness of the image decreases as the gas pressure rises, and the larger the diameter of the micro-holes, the faster the drop. In 1972, the previous technology used the same design to carry out a series of improvement and application tests to compare the thermal conductivity of sample support films of different materials such as graphite, carbon film, yttrium oxide and yttrium, and studied the catalytic catalysis of zinc, silver, nickel, iron and copper. The process of graphite oxidation.

Most of the above designs are replaced by a conventional penetrating electron microscope test piece stage, which is replaced by a modified environmental device, and is provided with a line for entering and leaving gas and entering and leaving the liquid to adjust the pressure difference between the environment device and the microscope vacuum chamber, regardless of It is a pore-limiting or film-limited environmental device. The advantage is that different gases can be used to react and adjust different humidity. The disadvantage is that the system is complicated and expensive, and it is necessary to change the hardware equipment (such as special test strip carrier, additional gas supply device, pressure sensing and control system, etc.). Such environmental electron microscopy tests have been applied in a variety of studies (such as dynamic chemical reactions, gas deposition of nanoparticles, microbial reduction of metals, gas phase formation of polypropylene and observation of copper oxidation by high voltage electron microscopy).

Similar environmental device concepts are also applied to the development of transmissive X-ray microscopes. The tantalum nitride film absorbs relatively little X-rays and is suitable as a penetrating film for environmental devices, and can be sealed with a rubber O-ring to seal a liquid sample. Another device for penetrating X-ray microscopy, which is compatible with the vacuum environment, has also been proposed. It also uses a tantalum nitride film on a metal test piece stage with a special pipeline and replaces the O-ring with a biocompatible glue. For sealing materials.

In particular, in 2000, the prior art proposed a two-piece tantalum substrate with a potassium hydroxide solution hollowed out, which can be directly placed in a conventional tantalum nitride film environment device without external piping, and applied to a vacuum. Scan through a penetrating X-ray microscope. Different from the mechanical precision machining of the past, it is a micro-electromechanical technology to make a completely enclosed enamel substrate environment device. The sample is limited to a thickness of about 500 nm between the two layers of tantalum nitride film. The liquid sample enters the environmental device by capillary force to form a liquid film, and the sample is sealed with epoxy resin and the two hollow substrates are bonded. The electron beam can pass through a relatively thin layer of tantalum nitride film and a liquid sample layer to obtain an electron microscope image of the wet sample. Since the amount of liquid between the films is very small, the liquid will dry out within a few tens of seconds.

In U.S. Patent No. 5,406,087, Japan JEOL Corporation proposes a film-sealed type environmental electron microscope design. This design is on a machined metal test strip stage using an O-ring to sandwich two layers of penetrating electronic film to create an airtight environment. Since the gas and liquid transfer lines in the stage pass into the environmental space, the moist biological sample can maintain the original type under normal pressure. A specially designed environmental-cell holder was installed on the transmission electron microscope of JEOL and FEI. The film uses a carbon film with poor mechanical strength, and pressure control equipment must be added to maintain operation.

U.S. Patent Publication No. 20070090289 discloses a method of observing a living biological sample in an electron microscope. The environmental device is an aperture-limited type space of a multi-layered space structure, the middle layer is a living environment of a biological sample, and at least one buffer layer is provided on the upper and lower sides, and the buffer layer is subjected to pressure and water vapor control by an external pipeline. In order to suppress the dissipation rate of the liquid and gas in the intermediate layer. There is still a need for additional pressure control equipment to precisely control the pumping and qi rates of the environmental unit.

The Republic of China Patent Publication No. 200826141 discloses a thin film-limited electron microscope environmental device made of a tantalum substrate. The film uses tantalum nitride to allow observation of live biological samples without changing the microscope hardware. The gas seal is filled with polymer epoxy resin, ultraviolet glue and silicone rubber, and a spacer is mixed therein to form an environmental space, and the sample enters the two films by capillary force. However, the adhesion of the two substrates of the above environmental device depends on the assistance of the optical observation system and the mobile platform, and the operation is complicated. Further, the airtightness of the polymer sealing material is not good, and it is easy to cause moisture absorption and gas leakage.

It is an object of the present invention to provide a sample carrying device for a non-contact electron microscope system which can be placed in any type of sample such as solid, liquid, gas, etc., particularly a sample that needs to be isolated from the outside, and allows electron beams, electromagnetic waves, light waves Physical quantities such as electric fields and magnetic fields penetrate the sample carrying device and act on the sample.

Another object of the present invention is to provide a self-aligning and automatic filling volume control design of a sample carrying device, which can reduce the drop in device yield due to misalignment and control the volume of the loaded material to reduce sample packaging and The complexity of environmental control.

Another object of the present invention is to provide a control technique for a sample holding space of a sample carrying device. The thickness of the sample receiving space can be controlled from 100 nm to several tens of micrometers to be suitable for samples of different sizes.

Another object of the present invention is to provide a structure of a sample carrying device to extend the path length of a sample liquid or gas leaking along a bonding interface to mitigate the occurrence of leakage.

Another object of the present invention is to provide a structure of a sample carrying device that increases the turning of the sample liquid or gas along the penetration path of the bonding interface and blocks the sealing material from penetrating into the cavity to avoid internal contamination.

Another object of the present invention is to provide a structure of a sample carrying device that prevents the combined gap and the sealing material from being directly irradiated by the electron beam, thereby prolonging the reliability and service life of the sample carrying device.

Another object of the present invention is to provide a sample carrying device that allows temperature control of a sample.

Another object of the present invention is to provide a sample carrying device that allows for fluid transport of a sample.

Another object of the present invention is to provide a gas seal filling technique for a sample carrying device, which effectively isolates the sample from the environment and the external environment, and the sealing process is simple and rapid.

Another object of the present invention is to provide a sample carrying device for use in electron irradiation, which effectively eliminates the effects of charge accumulation and temperature concentration, particularly under high energy and high current density electron beam bombardment.

SUMMARY OF THE INVENTION An object of the present invention is to provide a sample carrying device for an electron microscope and a method of fabricating the same, which utilizes a metal seal, thereby avoiding moisture absorption, leakage of liquid or leakage of vaporized gas. In addition, the sample carrying device of the electron microscope of the present invention and the manufacturing method thereof can align the cover member and the base of the present invention with each other without relying on the assistance of the optical observation system and the moving platform.

To achieve the above object, the present invention discloses a sample carrying device for an electron microscope, which comprises a cover member and a base. The cover member includes a body and a first film. The body includes a first side and a second side opposite to the first side, the first film is disposed on the second side, a groove is formed on the body and is open to the first side, the groove is exposed The first film extends horizontally to form a wing. The base includes a base portion, a soldering portion and a second film. The base portion includes an upper edge and a lower edge, the second film is disposed on the lower edge, a groove is formed in the base portion and opens on the upper edge, the groove exposes the second film, and the structure of the groove Corresponding to the structure of the body, the second film is aligned with the first film in a vertical direction, the soldering portion is disposed on the upper edge and corresponds to the wing portion, and the soldering portion is combined by a metal The object is welded to the cover member.

The invention further discloses a method for manufacturing a susceptor, comprising the steps of: chemical vapor deposition of a tantalum nitride layer on the lower edge and the upper edge of the base portion; etching the upper edge of the tantalum nitride layer to form a first opening, the maximum diameter of the first opening is defined as W1; etching the base portion and forming a first notch, the depth of the first notch is defined as D1; etching the tantalum nitride layer of the upper edge Forming a second opening, the maximum diameter of the second opening is defined as W2, wherein W2 is greater than W1; and etching the substrate portion to form the trench, wherein the trench includes a first trench and a first trench a second trench, the first trench receiving the body, the second trench exposing the second film, the depth of the first trench is defined as D2, and the thickness of the base portion is defined as T, T is less than or equal to D1 plus D2.

The technical features and advantages of the present disclosure have been broadly described above, and the detailed description of the present disclosure will be better understood. Other technical features and advantages of the subject matter of the claims of the present disclosure will be described below. It is to be understood by those of ordinary skill in the art that the present invention may be practiced otherwise. It is also to be understood by those of ordinary skill in the art that this invention is not limited to the spirit and scope of the disclosure as defined by the appended claims.

The subject discussed herein is a sample carrying device for an electron microscope and a method of making the same. In order to fully understand the present disclosure, detailed steps and structures will be set forth in the following description. Obviously, the implementation of the present disclosure is not limited to the specific details familiar to those skilled in the relevant art. On the other hand, well-known structures or steps are not described in detail to avoid unnecessarily limiting the disclosure. The preferred embodiments of the present disclosure will be described in detail below, but the disclosure may be widely practiced in other embodiments, and the scope of the disclosure is not limited, which is subject to the scope of the following patents. .

FIG. 1 is an exploded perspective view of the sample carrying device 10 of the present invention. The sample carrying device 10 can carry a sample that is easily damaged by a vacuum system in an electron microscope observation, such as a protein or a cell containing a liquid or water vapor, a solution of a nanoparticle, and the like. Based on the microelectromechanical technology, the sample carrying device 10 of the present invention can be directly placed into a micro-environment control device of a general electron microscope system. The sample carrying device 10 of the invention has the automatic alignment sealing function without the need of precision moving platform, high sealing yield, wide application range, strong electron bombardment tolerance, suitable for biological cell growth observation, and integration of liquid gas Replacement and temperature control functions, and the use of metal gas seals to prevent leakage of liquid gases.

As shown in the embodiment of FIG. 1, the sample carrying device 10 includes a cover member 110 and a base 120. The cover member 110 includes a body 111 and a first film 112. The thickness of the first film 112 is between 10 nm and 1 micron. The material of the first film 112 should be selected from amorphous yttrium nitride, aluminum nitride, yttrium oxide, aluminum oxide or the like. Film material, film thickness, film window size, window geometry, and ability to affect sample observation and airtight isolation. The thicker the film, the harder it is to penetrate the sample carrier 10 due to the physical quantity of electron beam, X-ray, etc., resulting in loss of image brightness and resolution; if the film is too thin, the mechanical strength is insufficient, and it cannot withstand the pressure difference between the inside and the outside, and is easily broken in a vacuum environment. , causing failure of the sample carrying device 10.

The body 111 includes a first side 113 and a second side 114 opposite the first side 113. The first film 112 is disposed on the second side 114. A groove 115 is formed in the body 111 and opens to the first side 113. The groove 115 exposes the first film 112. The groove 115 exposes the first film 112 to have an area of between 1 μm ² and 1 mm ² and the groove 115 exposes the shape of the first film 112 to a polygon of at least three sides, at least three sides herein. The above polygons contain circles and rectangles. The geometry of the exposed window of the first film 112, if there is a sharp angle, causes the stress concentration film to be easily broken at this point; if the window is round, smooth and has no sharp corners, the same pressure difference and the same film thickness can support larger The range film does not break.

As shown in FIG. 1 , the base 120 includes a base portion 121 , a solder portion 122 , and a second film 123 . The base portion 121 includes an upper edge 124 and a lower edge 125. The second film 123 is disposed on the lower edge 125. A groove 126 is formed in the base portion 121 and opens to the upper edge 124. The groove 126 exposes the bottom portion 124. The second film 123 has a structure corresponding to the structure of the body 111 such that the second film 123 is aligned with the first film 112 in the vertical direction. Therefore, the sample carrying device 10 of the present invention can be automatically aligned without using a precision moving platform having an automatic alignment sealing function to avoid a decrease in device yield due to misalignment. The groove 126 can accommodate part or all of the volume insertion of the body 111, and the cover member 110 and the base 120 can be combined in a manner to store the sample receiving space of the sample. The in-line mechanism provides the lateral positioning capability of the cover member 110 and the base 120 so that the film of the cover member 110 and the base 120 can be automatically aligned.

When the cover member 110 and the base 120 are automatically aligned and assembled, as shown in FIG. 2, the sample carrier 10 is viewed from above, since the cover member 110 and the base 120 are joined by soldering through the metal composition 130. The melting point of the metal composition ranges from 40 ° C to 500 ° C. 3 is a schematic view of the section line A-A of FIG. 2. As shown in FIG. 3, the first side 113 of the body 111 extends horizontally to form a wing portion 116. In other words, the cover member 110 has a laterally extending wing portion 116 with a tapered trapezoidal cone, and the first film 112 is located at the bottom of the trapezoidal cone. The soldering portion 122 is disposed on the upper edge 124 and corresponds to the wing portion 116, and the soldering portion 122 is soldered to the cover member 110 by the metal composition 130 to seal the cover member 110 and the base portion 120. In this embodiment, the first film 112 and the second film 123 form a sample accommodating space 140. The pitch of the combined films 112 and 123 is related to the depth of the sample accommodating space 140, so that the sample accommodating space 140 is provided. The thickness is limited to between 500 nm and 100 microns. If the sample receiving space 140 is too thick, the electron beam, X-ray, etc. are difficult to penetrate; if the thickness is too thin, there is not enough space for the sample such as cells and nanospheres to be in a movable natural state.

The occurrence of sample leakage is related to the speed and extent of diffusion of the sample liquid gas in the sample carrying device 10. In addition to selecting materials with low gas or liquid permeability, the length of the diffusion path can be increased to mitigate the occurrence of leakage. In the design, the cover member 110 has laterally extending wings 116. The extended slope of the cover member 110 and the two components of the base 120 allows the shortest distance between the sample receiving space 140 and the external vacuum environment to be more than 1.5 mm (doubled compared to the pure planar structure). In the three-dimensional structure, the path has two large angle transitions, which can effectively block the problem that the metal composition 130 penetrates into the sample receiving space 140 to cause internal pollution. The wing portion 116 formed by the extension of the body 111 can lengthen the path length of the sample liquid or gas leaking along the bonding interface of the cover member 110 and the susceptor 120, and increase the turning of the sample liquid or gas along the penetration path of the bonding interface. The wing 116 of the invention can slow the occurrence of sample liquid or gas leakage and block the metal composition 130 from penetrating into the sample receiving space 140 to avoid internal contamination.

As shown in the embodiment of Figures 4 to 6, the sample carrying device 10 of the present invention is suitable for biological cell attachment growth. The base 120 of the sample carrying device 10 is a dish-shaped tank. When in use, the liquid sample 150 is directly inserted (about 1 microliter, the liquid surface can be higher or lower than the groove opening), and can be used for placing cells such as cells and bacteria. growing up. When it is to be observed, it is combined with the cover member 110, and by the capillary force and the gravity, the cover member 110 slides into the groove 126 of the base 120 with the liquid level falling cover member, and the liquid sample is stored in the sample receiving space 140, and is completed. Automatic alignment. The excess culture solution is to be drained and sealed and placed in an electron microscope. A sample housing space 140 that is more suitable for long-term observation of a biological sample is provided than a similar product that must be injected through a flow channel opening.

7 to 11 are manufacturing flow charts of the cover member 110. Structure 20 includes a body layer 210 that is primarily comprised of a single crystal germanium substrate having a thickness of 250 microns. A layer of tantalum nitride 211, 212 is deposited on the upper and lower sides of the body layer 210 by low pressure chemical vapor deposition, and has a thickness of 20 nm to serve as an etching barrier for bulk micromachining, and is also a gas that allows electron beam and X-ray to penetrate. Closed film. An opening 220 is formed on the tantalum nitride layer 211 by a non-isotropic reactive ion etching (RIE). The opening 220 is then treated with a wet etch (potassium hydroxide solution) as shown in FIG. The openings 23, 240 are then formed by a non-isotropic reactive ion etching (RIE) of the tantalum nitride layer 212 as shown in FIG. The openings 220, 230, 240 are then treated by wet etching (potassium hydroxide) to form as shown in FIG. Finally, the cover member 110 of an embodiment of the present invention is completed by laser cutting. The second side 114 of the cover member 110 defines a projection 117 which is a tapered trapezoidal cone. In other embodiments, as shown in FIG. 12 and FIG. 13 , the openings 230 , 240 of the structure 20 ′ are coated with a metal layer 250 composed of chromium, nickel, copper, and gold (or referred to as a bump underlayer metal (UBM). )). In other words, the second side 114 other than the first film (ie, the tantalum nitride layer 212) is plated with a metal layer 250 of chromium, nickel, copper, gold to form another cover member 110'. Therefore, the protrusion 117 formed on the second side 114 of the body 111 is plated with a metal layer 250 composed of chrome, nickel, copper, and gold for soldering to the soldering portion 122 (refer to FIG. 1). Base 120. Welding materials should be selected from materials with low moisture absorption and high air tightness, such as non-polymer materials such as metal, glass and ceramics.

A manufacturing flow chart of the susceptor 120 is shown in FIGS. 14 to 17. The structure 30 includes a base portion 310 which is mainly composed of a single crystal germanium substrate having a thickness of 250 μm. A layer of tantalum nitride 311, 312 is deposited on the upper and lower sides of the base portion 310 by chemical vapor deposition, and has a thickness of 20 nm. A first opening 320 is formed on the tantalum nitride layer 311 by a non-isotropic reactive ion etching (RIE). The maximum diameter of the first opening 320 is defined as W1. Then, the first opening 320 is treated by wet etching (potassium hydroxide) to form a first notch 321 as shown in FIG. 15, and the depth of the first notch 321 is defined as D1. Then, as shown in FIG. 16, the tantalum nitride layer 311 is formed by a non-isotropic reactive ion etching (RIE) to form a second opening 330. The maximum diameter of the second opening 330 is defined as W2, where W2 is greater than W1. The second opening 330 is then treated by wet etching (potassium hydroxide) to form a trench 340 as shown in FIG. The trench 340 includes a first trench 341 and a second trench 342. The first trench 341 receives the body 111 of the cover member 110 (refer to FIG. 3), and the wing portion 116 contacts the base 120. The upper edge 124, here the upper edge 124, comprises a tantalum nitride layer 311. However, in various embodiments, as shown in FIG. 3, the upper edge 124 may not include the tantalum nitride layer 311 but only the base portion 121. Upper edge 124. In the embodiment of susceptor 120 of FIG. 17, second trench 342 exposes tantalum nitride layer 312 (ie, second film 123 of FIG. 3). In this embodiment, the first trench 341 has an area of 100 μm ² to 9 mm ² . The second trench 342 exposes the second film 312 to have an area of 1 μm ² to 4 mm ² , and the second trench 342 exposes the second film 312 to have a shape of at least three sides. Polygons above three sides contain circular, rectangular, etc., due to the special corner attack characteristics of the body micro-machining, gradually tend to a smooth bowl-shaped depression. When the most recessed point contacts the tantalum nitride film, a circular window is formed through the substrate, and as the etching depth increases, the window diameter increases. The time control of the two wet etches determines the depth of the environmental cavity and affects the thickness of the sample after the seal. The film window of the susceptor 120 exhibits a circular shape, reduces the stress concentration and breaks the yttrium nitride film, and can support a larger area than the square window without cracking, increasing the visual range of the environmental control device, and improving the window alignment error. Tolerance.

Referring to FIG. 3, the first film 112, the second trench 342, and the second film 123 form a sample receiving space 140. The volume of the sample receiving space 140 is 0.001 nanoliters (nl) to 10 milliliters ( Between ml). The distance between the first film 112 and the second film 123 is between 20 nanometers (nm) and 1 millimeter (mm). The thickness of the first film 112 and the second film 123 is between 10 nm and 1 μm.

In the embodiment of the other pedestal 120' of FIG. 18, the susceptor 120' of FIG. 18 is further provided with a metal mesh layer 350 under the yttrium nitride layer 312, as compared to the embodiment of FIG. The metal mesh layer 350 comprises chromium and gold. Under the electron bombardment of electron microscopy with high current density and high accelerating voltage, the material will have the problem of accumulated charge and radiant heat. Since the susceptor 120' has a specially designed conductive and thermally conductive metal mesh layer 350, it can be increased. The elimination of accumulated charges on the electron-transparent film (such as tantalum nitride layer 312) prevents the film from being damaged by mechanical strength due to charge accumulation and thermal stress. The metal mesh layer 350 having the functions of conduction, heat conduction, and mechanical strength increase is selected from a metal, ceramic, semiconductor, or the like. The metal mesh layer 350 is formed in a grid shape covering the electron penetrating film, and the hole allows the electron beam to pass through. The metal mesh layer 350 also plays the role of strengthening the mechanical properties of the electron penetrating film, and is subjected to the pressure difference between the inside and the outside of the sample carrying device. The resulting material is deformed to increase the life of the electron through the film. In the embodiment of the susceptor 120' of FIG. 19, the region of the solder portion 360 of the tantalum nitride layer 311 of the pedestal 120' may further form a heating circuit 370 (the heating circuit 370 may be composed of aluminum). As shown in FIG. 20, an insulating layer 380 is overlaid on the heating circuit 370. Finally, a bump bottom metal layer 390 is disposed on the insulating layer 380. The bump bottom metal layer 390 is composed of chromium, nickel, copper, and gold, and the topmost bump bottom metal layer 390 may be deposited with tin (Sn) and cadmium. (Cd) and indium (In) form a low melting temperature solder. The insulating layer 380 can assist in electrical insulation between the heating circuit 370 and the bump bottom metal layer 390, which is isolated from the current by the tantalum nitride. In this embodiment, the soldering portion 360 includes a heating circuit 370, an insulating layer 380, and a bump bottom metal layer 390. In other words, a heating circuit 370 and an insulating layer 380 are disposed at the bottom of the bump bottom metal layer 390, and the insulating layer 380 is disposed. Between the heating circuit 370 and the bump bottom metal layer 390. However, in other embodiments (not shown), the solder portion 360 may also include only the bump bottom metal layer 390 without the design of a heating circuit or insulating layer.

As shown in another embodiment of FIG. 22, the sample carrier 40 includes a cover member 410 and a base 420. The cover member 410 includes a body 411 and a first film 412 (which may also be a tantalum nitride layer). The body 411 includes a first side 413 and a second side 414 opposite the first side 413. The first film 412 is disposed on the second side 414. A first layer 413 is provided with a tantalum nitride layer 412'. The fabrication process of the first film 412 and the tantalum nitride layer 412' is as shown in the embodiment of FIGS. 7 to 11. As shown in FIG. 22, the cover member 410 further includes a metal mesh layer 415 disposed under the first film 412, which can also solve the material bearing device 40 under the electron bombardment of the electron microscope with high current density and high acceleration voltage. A problem of accumulated charge and radiant heat can occur, as shown in Figure 23, the metal mesh layer 415 contains a plurality of grids 418 for electron beam bombardment. In addition, in this embodiment, the bump bottom metal layer 416 plated on the second side 414 may also be limited to the wing portion 417 opposite to the solder portion 422. The bump bottom metal layer 416 is made of chromium and nickel. , copper, gold. In other embodiments (not shown), the heating circuit 470 can also be disposed at the bottom of the wing portion 417, that is, the bottom metal layer 416 of the bump. As shown in FIG. 22, the base 420 includes a base portion 421, a solder portion 422, and a second film 423. The material of the body 411 or the base portion 421 is a semiconductor or a metal oxide, the semiconductor is a double polished or single polished single crystal germanium, and the metal oxide is aluminum oxide. The first film 412 or the second film 423 is selected from the group consisting of cerium oxide, cerium nitride, and cerium carbide. In the embodiment shown in FIG. 22, the base portion 421 includes an upper edge 424 and a lower edge 425. The second film 423 is disposed on the lower edge 425. A groove 426 is formed on the base portion 421 and is open to the base portion 421. The upper edge 424 exposes the second film 423. The structure of the groove 426 corresponds to the structure of the body 411 to align the second film 423 with the first film 412 in the vertical direction.

In the embodiment shown in FIG. 22, the solder portion 422 includes a heating circuit 470, an insulating layer 480, and a bump bottom metal layer 490. The heating circuit 470 can heat the bump bottom metal layer 490 to be soldered to the bump bottom metal layer 416 to seal the cover member 410 and the base 420 of the sample carrier 40, so that the first film 412 and the second film 423 form a sample. The accommodation space 427. When the metal material is used for sealing, the heating circuit 470 and the under bump metallization 490 (UBM) can heat the molten metal sealing material in a very short time to make the bump bottom layer of the cover member 410. The metal 416 creates a bond that still ensures that the sample is not affected by thermal shock. The heating circuit 470 can also be made of metal or polysilicon. The thinner wire heating circuit 470 is located on a film window (not shown), and the heating circuit 470 is electrically connected to the two electrodes. When the heating circuit 470 is used for temperature rise control, a current is input from the two electrodes, the heating circuit 470 is heated by the metal to warm the sample under the film; and the resistance of the two electrodes is measured along with the heating current to match the heating obtained by the experiment. The circuit 470 has a Temperature Coefficient of Resistance (TCR), which can instantly convert the temperature of the heating circuit 470 to achieve temperature monitoring.

In addition, as shown in FIG. 24, a heat insulating groove 428 is disposed between the heating circuit (not shown) and the sample accommodating space 427 to prevent the heating circuit from conducting heat to the sample accommodating space 427. In the embodiment shown in Figure 24, a microchannel 429 is additionally included for fluid to enter and exit the sealed sample carrier or to increase the space for liquid storage. Various liquid gases can be replaced to change environmental conditions such as humidity and ion concentration; and liquid or gas to be involved in the reaction can be introduced at different times, and reactions such as catalysis, precipitation, and growth after mixing can be observed. The formation of microchannels can be performed on the tantalum substrate by wet chemical etching or dry plasma assisted ion etching. By means of vapor deposition of metal, chemical vapor deposition of tantalum nitride film or coating of polymer, yellow light lithography and etching can be used to make microchannels 429 with a depth of less than 200 microns, allowing fluid transport to the sample.

A method for fabricating a susceptor according to the present invention as shown in FIG. 25, comprising the steps of: step 2510, chemical vapor deposition of a tantalum nitride layer on the lower edge and the upper edge of the base portion; and step 2520, etching the a first opening is formed in the upper edge of the tantalum nitride layer, and a maximum diameter of the first opening is defined as W1; in step 2530, the base portion is etched to form a first notch, and the depth of the first notch is defined D1; step 2540, etching the tantalum nitride layer of the upper edge to form a second opening, the maximum diameter of the second opening is defined as W2, wherein W2 is greater than W1; and step 2550, etching the base portion Forming the trench, wherein the trench includes a first trench and a second trench, the first trench receiving the body, the second trench exposing the second film, the first trench The depth is defined as D2, and the thickness of the base portion is defined as T, and T is less than or equal to D1 plus D2.

Another method for fabricating a susceptor according to the present invention as shown in FIG. 26 includes the following steps: Step 2610, chemical vapor deposition of a tantalum nitride layer on the lower edge and the upper edge of the substrate portion; and step 2620, etching The upper edge of the tantalum nitride layer forms a first opening, the maximum diameter of the first opening is defined as W1; in step 2630, the base portion is etched to form a first notch, the depth of the first notch Defined as D1; in step 2640, a metal layer is deposited on the lower edge, the metal layer comprises chromium and gold; and in step 2650, the tantalum nitride layer of the upper edge is etched to form a second opening, the second opening The maximum path length is defined as W2, where W2 is greater than W1; in step 2660, the base portion is etched and the trench is formed, wherein the trench includes a first trench and a second trench, the first trench is received The second trench exposes the second film, the depth of the first trench is defined as D2, and the thickness of the base portion is defined as T, T is less than or equal to D1 plus D2; and step 2670 forms the soldering And a heating circuit on the upper edge, wherein the heating circuit is formed on the upper edge of the nitriding Layer, an insulating layer covering the heating circuit after forming a metal bump layer on the insulating layer.

Implementation example:

According to the sample carrying device and the sealing technology according to the embodiment of the invention, the substrate is a single crystal germanium, which can be chemically etched with potassium hydroxide or tetramethylammonium hydroxide to form a groove with a specific angle of slope. The above-mentioned film is a non-crystalline tantalum nitride film, and a low-stressed tantalum nitride film grown by low-pressure chemical vapor deposition has sufficient strength at a limited thickness to support a pressure difference of more than one atmosphere, and does not cause an electron beam due to thickness. Or the penetration intensity of X-rays is reduced too much. The metal mesh layer is made of gold, and is grown by an electron gun vapor deposition machine and deposited by physical vapor deposition. The good conductivity and heat transfer capability of the metal mesh layer can quickly derive the charge and heat caused by electron beam bombardment. . On the other hand, when the sealing material is the bottom metal layer of the bump, the metal layer at the bottom of the bump also serves as a wet contact point of the molten metal sealing material. The above-mentioned sealing material is a low melting point alloy containing a metal such as bismuth, plumbum, tin (Stannum), cadmium or indium, and is sealed at a temperature rising and melting state. Filling, cooling and returning to solid form a high-strength barrier barrier for liquids and gases. The melting point temperature is between 40 ° C and 300 ° C. The melting point is too high. The temperature gradient in the melting process affects the sample. The melting point is too low. The sealing material will melt due to temperature changes caused by electron beam bombardment of the environmental device.

The embodiment of the invention is an environmental device design based on MEMS technology, which can mass-produce the disposable sample carrying device, which is cheap and has no pollution problem of repeated use. The sample carrying device is suitable for both high-vacuum transmission electron microscope and atmospheric X-ray microscope. After the filling is completed, it can be directly placed in the penetration electron microscope test piece for observation without any hardware modification. It is possible to upgrade a general penetrating electron microscope to an environmental penetrating electron microscope.

In the sample carrying device of the embodiment of the invention, the cover member has an outer diameter of 1.5 mm and a base outer diameter of 2.8 mm. Both the cover and the base are etched from the 250 micron thick tantalum substrate by micromachining. The cover member has a square electron beam with a side length of 100 μm, and the base has a circular electron beam of 300 μm in diameter, which covers a 20 nm thick tantalum nitride film. The film window area of the pedestal has a metal mesh to remove charge build-up and heat and increase the mechanical strength of the film, with a grid spacing of about 30 microns. The liquid-containing sample is stored between the sample holding spaces and has a thickness of 20 μm, which is suitable for observation of bacteria of several micrometers. When used in an electron microscope, the lower edge of the pedestal faces upwards and is subjected to electron beam irradiation; while the upper edge of the pedestal is hidden under the lower rim of the larger pedestal to protect it from electron beam bombardment. The electron beam passes through the metal mesh and penetrates the sample in the sample housing space and is imaged on the microscope image plane.

The technical content and the technical features of the present disclosure have been disclosed as above, but those skilled in the art should understand that the teachings and disclosures of the present disclosure are disclosed without departing from the spirit and scope of the disclosure as defined by the appended claims. Can be used for various substitutions and modifications. For example, many of the devices or structures or method steps disclosed above may be implemented in different ways or substituted with other structures, or a combination of the two.

The scope of the present invention is not limited to the process, machine, manufacture, compositions, means, methods, or steps of the particular embodiments disclosed. It should be understood by those of ordinary skill in the art that, based on the teachings of the present disclosure, the process, the machine, the manufacture, the composition of the material, the device, the method, or the steps, whether present or future developers, The revealer performs substantially the same function in substantially the same manner, and achieves substantially the same result, and can also be used in the present disclosure. Accordingly, the scope of the following claims is intended to cover such <RTIgt; </ RTI> processes, machines, manufactures, compositions, devices, methods or steps.

10. . . Sample carrier

110. . . Cover

110'. . . Cover

111. . . Ontology

112. . . First film

113. . . First side

114. . . Second side

115. . . Groove

116. . . Wing

117. . . Protrusion

120. . . Pedestal

120'. . . Pedestal

121. . . Base body

122. . . Welding department

123. . . Second film

124. . . Upper edge

125. . . Lower edge

126. . . Trench

130. . . Metal composition

140. . . Sample accommodation space

150. . . Liquid sample

20. . . structure

20'. . . structure

210. . . Body layer

211. . . Tantalum nitride layer

212. . . Tantalum nitride layer

220. . . Opening

230. . . Opening

240. . . Opening

250. . . Metal layer

30. . . structure

310. . . Base body

311. . . Tantalum nitride layer

312. . . Tantalum nitride layer, second film

320. . . First opening

321. . . First gap

330. . . Second opening

340. . . Trench

341. . . First groove

342. . . Second groove

350. . . Metal mesh layer

360. . . Welding department

370. . . Heating circuit

380. . . Insulation

390. . . Bump bottom metal layer

40. . . Sample carrier

410. . . Cover

411. . . Ontology

412. . . First film

412'. . . Tantalum nitride layer

413. . . First side

414. . . Second side

415. . . Metal mesh layer

416. . . Bump bottom metal layer

417. . . Wing

418. . . grid

420. . . Pedestal

421. . . Base body

422. . . Welding department

423. . . Second film

424. . . Upper edge

425. . . Lower edge

426. . . Trench

427. . . Sample accommodation space

428. . . Insulation tank

429. . . Microchannel

470. . . Heating circuit

480. . . Insulation

490. . . Bump bottom metal layer

1 shows an exploded schematic view of a sample carrying device in accordance with an embodiment of the present invention;

Figure 2 shows a top view of the sample carrier assembly in accordance with one embodiment of the present invention;

Figure 3 shows a cross-sectional view of the sample carrying device according to section line A-A of Figure 2;

4 to 6 are schematic views showing the use of a package of a sample carrying device according to an embodiment of the present invention;

7 to 11 are schematic views showing a manufacturing process of a cover member according to an embodiment of the present invention;

12 to 13 are schematic views showing a manufacturing process of a cover member according to another embodiment of the present invention;

14 to 17 are schematic views showing a manufacturing process of a susceptor according to an embodiment of the present invention;

18 to 21 are schematic views showing a manufacturing process of a susceptor according to another embodiment of the present invention;

Figure 22 shows a side view of a sample carrying device in accordance with yet another embodiment of the present invention;

Figure 23 is a view showing a cover body according to still another embodiment of the present invention;

Figure 24 shows a top view of a susceptor in accordance with still another embodiment of the present invention;

Figure 25 is a flow chart showing a method of manufacturing a susceptor according to an embodiment of the present invention;

Figure 26 is a flow chart showing a method of fabricating a susceptor in accordance with another embodiment of the present invention.

40. . . Sample carrier

410. . . Cover

411. . . Ontology

412'. . . Tantalum nitride layer

413. . . First side

414. . . Second side

415. . . Metal mesh layer

416. . . Bump bottom metal layer

417. . . Wing

420. . . Pedestal

421. . . Base body

422. . . Welding department

423. . . Second film

424. . . Upper edge

425. . . Lower edge

426. . . Trench

427. . . Sample accommodation space

470. . . Heating circuit

480. . . Insulation

490. . . Bump bottom metal layer

Claims (22)

A sample carrying device for an electron microscope, comprising: a cover member comprising a body and a first film, wherein the body comprises a first side and a second side opposite to the first side, the first film Provided on the second side, a recess is formed in the body and opens on the first side, the recess exposes the first film, the first side extends horizontally to form a wing portion; and a base includes a base body a portion, a soldering portion and a second film, wherein the base portion includes an upper edge and a lower edge, the second film is disposed on the lower edge, a groove is formed in the base portion and is open to the upper edge, the groove The groove exposes the second film, and the structure of the groove corresponds to the structure of the body, so that the second film is aligned with the first film in a vertical direction, and the soldering portion is disposed on the upper edge and corresponds to The wing portion is welded to the cover member by a metal composition. The sample carrying device of claim 1, wherein the second side of the body forms a protrusion, and the second side of the first film is plated with a metal composed of chromium, nickel, copper and gold. Floor. The sample carrying device of claim 1, wherein the groove comprises a first groove and a second groove, the first groove receiving the body, the wing contacting the upper edge, the second The trench exposes the second film. The sample carrying device according to claim 3, wherein the first groove has an area of from 100 μm ² to 9 mm ² . The sample carrying device according to claim 3, wherein the second trench exposes the second film to have an area of 1 μm ² to 4 mm ² , and the second trench exposes the second film to have a shape of at least three sides. Polygon. The sample carrying device according to claim 3, wherein the first film, the second groove and the second film form a sample accommodating space, and the sample accommodating space has a volume of 0.001 liters (nl) to 10 Milliliter (ml). The sample carrying device according to claim 6, wherein a distance between the first film and the second film is from 20 nanometers (nm) to 1 mm (mm). The sample carrying device according to claim 1, wherein the groove exposes the first film to have an area of from 1 μm ² to 1 mm ² . The sample carrying device according to claim 1, wherein the groove exposes the shape of the first film to be at least three or more polygons. The sample carrying device according to claim 1, wherein the first film and the second film have a thickness of 10 nm to 1 μm. The sample carrying device according to claim 1, wherein the metal composition has a melting point ranging from 40 ° C to 500 ° C. The sample carrying device according to claim 1, wherein the welded portion is a bump bottom metal layer, and the bump bottom metal layer is composed of chromium, nickel, copper, and gold. The sample carrying device according to claim 1, wherein the wing portion corresponding to the welded portion is a bump bottom metal layer, and the bump bottom metal layer is composed of chromium, nickel, copper, and gold. The sample carrying device of claim 12 or 13, wherein a heating circuit is disposed at a bottom of the bump bottom metal layer. The sample carrying device of claim 14, wherein the first film and the second film form a sample receiving space, and the heating circuit and the sample receiving space are provided with a heat insulating groove to prevent the heating circuit from conducting heat to Sample accommodation space. The sample carrying device according to claim 1, wherein the material of the body or the base portion is a semiconductor or a metal oxide, the semiconductor is a double polished or single polished single crystal germanium, and the metal oxide is aluminum oxide. The sample carrying device of claim 1, wherein the first film or the second film is selected from the group consisting of ceria, tantalum nitride, and tantalum carbide. A method of manufacturing the susceptor in the sample carrying device of claim 1, comprising the steps of: chemical vapor deposition of a tantalum nitride layer on the lower edge of the base portion and the upper edge; etching the a first opening is formed by the upper edge of the tantalum nitride layer, the maximum diameter of the first opening is defined as W1; the base portion is etched and a first notch is formed, the depth of the first notch is defined as D1; Etching the tantalum nitride layer of the upper edge to form a second opening, the maximum diameter of the second opening is defined as W2, wherein W2 is greater than W1; and the base portion is etched and the trench is formed, wherein the trench The trench includes a first trench and a second trench, the first trench accommodates the body, the second trench exposes the second film, the depth of the first trench is defined as D2, and the base portion The thickness is defined as T, and T is less than or equal to D1 plus D2. The method of claim 18, further comprising depositing a metal layer on the lower edge, the metal layer comprising chromium and gold. The manufacturing method according to claim 18, further comprising forming the soldering portion and a heating circuit on the upper edge. The manufacturing method according to claim 20, wherein the heating circuit is formed on the tantalum nitride layer of the upper edge. The manufacturing method according to claim 21, wherein after covering an insulating layer on the heating circuit, a bump bottom metal layer is formed on the insulating layer.