JP2005062130A - Small flake production equipment - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an apparatus capable of sampling an arbitrary part of a volatile substance-containing sample such as an organic matter or a living organism as a minute thin piece sample having a size capable of being observed with a transmission electron microscope without being altered, and simultaneously capable of being observed with an electron microscope. To do.
SOLUTION: A sample chamber 2 capable of controlling the atmosphere, a sample holder 14 for introducing a volatile substance-containing sample into the sample chamber, a scanning electron microscope for observing the held sample, and held by the sample holder 14 A manipulator for sampling the sample and a sample holder 15 for taking the sampled sample out of the sample chamber 2 are provided. With this configuration, the volatile substance-containing sample can be sampled without being altered, and can be easily produced as an observation sample for a transmission electron microscope.
[Selection] Figure 1

Description

  The present invention relates to a micro flake production apparatus for producing a micro flake of a volatile substance-containing sample such as an organic substance or a living organism.

  Conventionally, various methods have been devised as sample preparation methods for transmission electron microscope observation. In particular, the microtome method is used for biological samples and polymer samples. In the microtome method, after embedding the target sample in a resin such as epoxy or araldite, an ultrathin section is prepared using a glass knife or diamond knife, and the section suspended in water is scooped up with a grid, and a sample for transmission electron microscope observation (See Non-Patent Document 1).

  On the other hand, there is a FIB (Focused Ion Beam) processing method as a method of forming a sample cross section of a specific part with a spatial resolution of submicron order. The FIB processing method is a method in which Ga ions are focused on a desired specific region and only that region is sputter-etched (see Non-Patent Document 2).

  Furthermore, as a method of peeling or cutting a part of the sample and moving it to a predetermined place, it is conceivable to use a manipulator. In particular, a method using a manipulator is effective when preparing a sample for observation with a transmission electron microscope. When operating a manipulator, a method of observing the operation state is important. Generally, an optical microscope (Japanese Patent Laid-Open No. 11-271036, etc.), a scanning electron microscope (Japanese Patent Laid-Open No. 2001-198896, Japanese Patent Laid-Open No. 2001) are used. -88100 publication etc.) etc. are used in combination.

In addition, in morphological observation of these biological samples and polymer samples with an electron microscope, stereoscopic observation by high-tilt observation is necessary in order to observe the outer shape and internal structure.
Kentaro Asakura and Yasuhisa Hirohata "Ultra-microtome technique Q &A" published by Agne Jofusha, 1999, September 30 Authored by Masao Hirasaka and Kentaro Asakura “FIB, AEON MILLING TECHNOLOGY Q & A” Agne Jofusha Publishing 2002/9/25

  However, since the microtome method as the conventional technique is a slicing method, only one section of the sample can be observed. Further, in order to observe a target site, there is only a method of approaching the site while repeating preparation of a section and observation with a transmission electron microscope. In addition, the compatibility between the sample and the embedding resin, such as hardness, adhesion, machinability, and chemical influence, is very important, and trial and error are indispensable methods.

  On the other hand, when an organic material is processed using the FIB processing method, the processing damage is larger than that of a metal material or a semiconductor material. Furthermore, since the sample must be introduced into a vacuum environment, the sample containing a volatile substance is deformed and altered, and cannot be used.

  Further, although an observation method is used together when operating the manipulator, the optical microscope has insufficient spatial resolution in order to produce a microscopic thin piece that can be observed with a transmission electron microscope. In addition, when a scanning electron microscope is used in combination, the spatial resolution is sufficient, but since the sample must be introduced into a vacuum environment, the sample containing a volatile substance is deformed and altered, and cannot be used.

  In addition, in order to obtain information on the three-dimensional outline and internal structure of these biological samples and polymer samples, it is necessary to perform surface morphology observation using a scanning electron microscope and transmission image observation using a transmission electron microscope from various directions. However, in the prior art, each electron microscope must be observed separately, and a scanning electron microscope and a transmission electron microscope apparatus capable of high-tilt observation are necessary for observation from various directions.

  The present invention has been made in view of the above-mentioned conventional problems, and its purpose is to have a size that enables observation with a transmission electron microscope without altering any part of a volatile substance-containing sample such as organic matter or organisms. An object of the present invention is to provide a micro thin piece manufacturing apparatus that can be easily manufactured as a micro thin piece.

  In order to achieve the above object, the present invention observes a sample chamber capable of atmospheric control, a first sample holder for introducing a volatile substance-containing sample into the sample chamber, and a sample held in the sample holder. A scanning electron microscope for sampling, a manipulator for sampling the sample held in the first sample holder, and a second for holding the sample sampled by the manipulator and taking it out of the sample chamber And a sample holder.

  According to this configuration, the present invention can be easily manufactured as a minute flake of a size that can be observed with a transmission electron microscope without altering an arbitrary part of a volatile substance-containing sample such as an organic substance or a living organism.

  Further, the present invention is characterized in that, in the apparatus configured as described above, the manipulator is temperature-controllable and includes an electron detector for observing a sampled sample. It is an electron detector for observing a microscope image and an electron detector for observing a bright field image and a dark field image of a scanning transmission electron microscope image.

  In the present invention, the inside of the sample chamber is evacuated to such a degree that the sample does not deteriorate, and the manipulator is operated while observing with a scanning electron microscope, and sampling is performed using a small thin piece of the target sample as a sample for observation with a transmission electron microscope. Do. Furthermore, after cooling the sample by the temperature control mechanism of the manipulator and evacuating the lens chamber and sample chamber in the electron optical column, the surface of the minute slice is irradiated with an electron beam and scanned to detect electrons for a scanning electron microscope. The secondary electron image can be observed with a detector and the scanning transmission electron microscope image can be observed with an electron detector for a scanning transmission electron microscope. Furthermore, it is possible to simultaneously perform tilt observation by rotating the needle shaft.

  Therefore, according to this configuration, the present invention can easily produce an arbitrary part of a volatile substance-containing sample such as an organic substance or a living organism as a small thin piece having a size that can be observed with a transmission electron microscope without degeneration, It is possible to easily observe the external shape or internal structure of the sample on the spot with an electron microscope.

  According to the present invention, an arbitrary part of a volatile substance-containing sample such as an organic substance or a living organism can be easily sampled as a small thin piece having a size that can be observed with a transmission electron microscope without damaging the sample. In addition, any part of a volatile substance-containing sample such as organic matter or living organisms can be easily sampled as a small thin piece of a size that can be observed with a transmission electron microscope without damaging the sample. Thus, the sampled micro thin sample can be observed with an electron microscope.

  Next, the best mode for carrying out the present invention will be described in detail with reference to the drawings.

(First embodiment)
FIG. 1 is a block diagram showing a first embodiment of the present invention. In the figure, 1 is an electron optical column, and 2 is a sample chamber (sample chamber) for introducing a volatile substance-containing sample. Inside the electron optical column 1, an electron gun chamber 4 in which an electron gun 3 is installed, a condenser lens 5, a scanning coil 6, an astigmatism correction device 7, and a lens chamber 9 having an objective lens 8 are provided.

  Further, inside the sample chamber 2, a needle 12, a sample holder 14, and a transmission electron microscope sample holder 15 are installed. The needle 12 is connected to the tip of a manipulator (not shown) that can be operated from the outside. The needle 12 is made of, for example, metal or glass coated with metal.

  The sample holder 14 holds a sample 13 containing a volatile substance such as a living organism or a polymer and introduces the sample into the sample chamber 2. The sample holder 15 for the transmission electron microscope is a sample peeled off from the sample 13 or a sample. A sample cut from 13 is held and taken out of the sample chamber 2 as a sample for observation with a transmission electron microscope. The sample holder 14 and the transmission electron microscope sample holder 15 each have a mechanism that can be individually introduced through the preliminary exhaust chambers 20 and 21 that have been exhausted in advance to a predetermined degree of vacuum. The preliminary exhaust chamber 20 is evacuated to a predetermined degree of vacuum before introducing the sample, and the preliminary evacuation chamber 21 is evacuated to a predetermined degree of vacuum before discharging the sample.

  The electron gun chamber 4 in the electron optical column 1 is connected from the exhaust unit 16 to an exhaust system (not shown), and the lens chamber 9 is connected from the exhaust unit 17 to an exhaust system (not shown). It is possible to maintain the chamber vacuum. On the other hand, the sample chamber 2 is connected to an exhaust system (not shown) from the exhaust unit 18 and is provided with a gas introduction unit 19 through a valve, so that an arbitrary gas atmosphere (for example, water, nitrogen, etc.) can be used. It is possible to maintain the degree of vacuum.

  Reference numerals 10 and 11 denote orifices that partition regions having different pressures and allow an electron beam to pass therethrough. The orifice 10 partitions the electron gun chamber 4 and the lens chamber 9, and the orifice 11 partitions the lens chamber 9 and the sample chamber 2. Usually, an objective aperture is used as the orifice 11.

Next, a method for preparing a minute thin piece sample of a volatile substance-containing sample will be described. First, the degree of vacuum in each of the electron gun chamber 4, the sample chamber 2, and the lens chamber 9 is adjusted. Since the electron gun chamber 4 needs to generate an electron beam by the electron gun 3, the electron gun chamber 4 is exhausted from the exhaust unit 16 and adjusted to a vacuum degree of about 10 ⁻³ Pa.

  On the other hand, the sample chamber 2 is adjusted to a degree of vacuum so that a sample containing a volatile substance such as an organic substance or a living organism can be introduced, that is, the volatile substance-containing sample is not altered. In this method, first, the exhaust unit 18 is evacuated so that the degree of vacuum is 1 Pa or less, and then a gas such as water or nitrogen is introduced from the gas introduction unit 19 to adjust the degree of vacuum to about 1000 Pa. The lens chamber 9 located between the sample chamber 2 and the electron gun chamber 4 is evacuated from the exhaust section 17 and adjusted to a degree of vacuum of about 1 Pa.

  Next, the sample holder 14 carrying the sample 13 is introduced into the sample chamber 2 through the preliminary exhaust chamber 20 and placed so that the sample 13 is positioned on the optical axis of the electron beam of the electron gun 3. Further, although the transmission electron microscope sample holder 15 is introduced through the preliminary exhaust chamber 21, it is placed in a retracted position so as not to contact the sample holder 14.

  Next, electrons are emitted from the electron gun 3, and the condenser lens 5, the astigmatism correction device 7, and the objective lens 8 are operated to irradiate a finely focused electron beam on the sample 13. Further, the scanning coil 6 is driven to deflect the electron beam, and the electron beam is scanned on the surface of the sample 13. At this time, the sample current of the sample 13 is measured by a detector (not shown), and the measured current and a signal for scanning the electron beam are displayed in synchronization with each other, thereby obtaining an image of the surface of the sample.

  The sample current is a current that flows between the sample 13 and the ground when the primary electron beam is incident on the sample 13, and is the amount obtained by subtracting electrons such as secondary electrons and reflected electrons emitted from the sample 13 from the primary electron beam. Current.

  In this apparatus, the current can also be measured from the needle 12 and the transmission electron microscope sample holder 15, and the current of the sample 13, the needle 12, and the transmission electron microscope sample holder 15 is detected for image display. Each of them is detected by a device (not shown). Then, by displaying the detected current of each part and the signal for scanning the electron beam in synchronization with each other, an image in which the sample 13, the needle 12, and the sample holder 15 are combined can be obtained. In this case, since the positional relationship among the sample 13, the needle 12, and the sample holder 15 is known, operability is improved.

  Next, the surface of the sample 13 is focused, and the part of the sample 13 to be sampled is determined. Thereafter, the manipulator is operated to bring the needle 12 closer to the sample 13 and peel off a desired portion of the sample 13. In some cases, it is also possible to cut a part of the sample 13 using the two needles 12. Finally, a portion to be used as a sample of the transmission electron microscope is attached to the needle 12 and retracted from the surface of the sample 13 to the electron optical column 1 side.

  Further, the sample holder 14 is retracted to the preliminary exhaust chamber 20 side so that the sample 13 does not contact the needle 12, and the transmission electron microscope sample holder 15 is moved on the optical axis of the electron beam. At this time, if the transmission electron microscope sample holder 15 is moved and focused without changing the setting of each lens, the sample holder 15 can be prevented from contacting the needle 12.

  Next, the manipulator is operated again to bring the needle 12 close to the mesh portion of the transmission electron microscope sample holder 15 (the sample holding portion at the time of observation with the transmission electron microscope), and a part of the sample sampled earlier is placed. When it is difficult to separate the sample from the needle 12, another needle 12 may be used. After separating the needle 12 from the transmission electron microscope sample holder 15, the transmission electron microscope sample holder 15 is withdrawn from the sample chamber 2 through the preliminary exhaust chamber 21. This sample is used as a micro thin sample for a transmission electron microscope.

  In the present embodiment, an arbitrary part of a volatile substance-containing sample such as an organic substance or a living organism can be sampled as a minute flake having a size that can be easily observed with a transmission electron microscope without being altered.

(Second Embodiment)
FIG. 2 is a block diagram showing a second embodiment of the present invention. The difference from the first embodiment is that a backscattered electron detector 22 for detecting backscattered electrons generated from the sample 13, the needle 12, and the transmission electron microscope sample holder 15 is provided below the electron optical column 1. There are other configurations similar to those in FIG. In this embodiment, when forming an image of the sample 13, the needle 12, or the transmission electron microscope sample holder 15, the signal detected by the backscattered electron detector 22, not the sample current, is synchronized with the signal for scanning the electron beam. The image is obtained by displaying the image.

  In this embodiment, since the image of the sample 13, the needle 12, and the transmission electron microscope sample holder 15 is displayed using the signal detected by the backscattered electron detector 22, glass or the like can be used as the material of the needle 12. The configuration of the present embodiment is effective when the metal needle cannot be used depending on the target volatile substance-containing sample.

  In the above embodiment, the needle type is used as the tip shape of the manipulator. However, the present invention is not limited to this, and for example, a scissor type or a vacuum suction nozzle using working exhaust may be used. Further, when the target sample is small, carbon nanotubes may be used. Further, although the operation is performed so that the focus is adjusted to the sample, a method in which the focus position is fixed to the tip of the manipulator and the focus is automatically corrected in conjunction with the movement of the needle may be used.

(Third embodiment)
FIG. 3 is a block diagram showing a third embodiment of the present invention. In FIG. 3, the same parts as those in FIGS. 1 and 2 are denoted by the same reference numerals. First, in this embodiment, mainly a sample chamber (sample chamber) 2 capable of controlling the atmosphere, an electron optical column 1, a manipulator 30 having a needle at the tip, an electron detector 31 for a scanning electron microscope, a scanning transmission It comprises an electron detector 32 for an electron microscope.

  In this embodiment, a minute thin piece sample 34 is sampled from a sample 13 containing a volatile substance such as a living organism or a polymer by a needle 35 at the tip of a manipulator 30 and scanned by an electron detector 31 for a scanning electron microscope as will be described in detail later. This is configured to enable observation with an electron microscope, or observation with a scanning transmission electron microscope using an electron detector 32 for a scanning transmission electron microscope.

  The electron optical column 1 includes an electron gun chamber 4 in which an electron gun 3 is installed, and a lens chamber 9 in which a condenser lens 5, a scanning coil 6, an astigmatism correction device 7, and an objective lens 8 are installed. The electron optical column 1 is partitioned by an orifice 10 installed between an electron gun chamber 4 and a lens chamber 9, and the electron gun chamber 4 and the lens chamber 9 are exhausted from exhaust units 16 and 17, respectively (not shown). ) To maintain a vacuum.

  In the sample chamber 2, a manipulator 30 having a needle 35 at the tip, a manipulator stage 36 that moves the manipulator 30 in the XYZ (see FIG. 3) direction, a guide rail 37 that guides the needle 35 of the manipulator 30 in the Φ direction, and a sample A sample stage 38 that is moved in the XYZΘ direction (see FIG. 3) is provided. The Φ direction is a circumferential direction around the sample stage 38 and is a rotation axis orthogonal to the Θ axis.

  In the sample chamber 2, an electron detector 31 for a scanning electron microscope that can be used as a secondary electron detector and a backscattered electron detector by an energy filter (not shown), a detection area for bright field observation, and a detection for dark field observation An electron detector 32 for a scanning transmission electron microscope divided into two with an area is installed. By using an energy filter, the electron detector 31 can separately detect secondary electrons from the sample (electrons emitted from the sample by irradiating the sample with an electron beam) and reflected electrons reflected from the sample. Is possible.

  Further, the sample chamber 2 is connected to an exhaust system (not shown) from the exhaust unit 18 and is provided with a gas introduction unit 19 through a valve, and can be maintained at an arbitrary vacuum degree in an arbitrary gas atmosphere. Mechanism. Further, the sample chamber 2 is connected to the sample introduction chamber 39 for introducing the sample 13 and the above-described electron optical column 1. At the tip of the electron optical column 1, an orifice 11 for an objective aperture is installed, and the sample chamber 2 and the electron optical column 1 are separated by the orifice 11.

  The manipulator 30 is configured such that a needle 35 connected to the tip of the manipulator 30 is driven in an XYZ direction by a manipulator stage 36 using a stepping linear motor and a piezoelectric element (not shown), and can further move in the Φ direction along the guide rail 37. is there. As described above, the needle 35 of the manipulator 30 can be freely operated from the outside, and the volatile substance-containing sample can be sampled.

  The manipulator 30 includes a temperature control mechanism 40 that controls the temperature of the needle 35 using a Peltier element (not shown) or the like. The needle 35 is connected to the tip of the manipulator 30 that can be operated from the outside as described above, and is made of metal or glass coated with metal. The temperature control mechanism 40 is used to cool the sample at the tip of the needle using a Peltier element.

Next, the sampling method of the micro thin sample according to the present embodiment will be described with reference to FIGS. First, the degree of vacuum at each part is adjusted. The electron gun chamber 4 is adjusted to a degree of vacuum of about 10 ⁻³ Pa because it is necessary to generate an electron beam by the electron gun 3. The sample chamber 2 contains a sample 13 containing a volatile substance such as an organic substance or a living organism by evacuating from the exhaust unit 18, introducing a desired gas (for example, water, nitrogen, etc.) from the gas introduction unit 19 and flow control. The vacuum degree is adjusted to about 1000 Pa so that the sample can be introduced (so that the sample does not deteriorate). The lens chamber 9 located between the sample chamber 2 and the electron gun chamber 4 is adjusted to a degree of vacuum of about 1 Pa.

  Next, in order to sample a minute thin piece sample, the volatile substance-containing sample 13 is introduced into the sample chamber 2 and observed with a scanning electron microscope. In order to introduce the sample 13, the sample stage 38 on which the sample 13 is mounted is introduced into the sample chamber 2 through the sample introduction chamber 39 evacuated to a predetermined degree of vacuum in advance. It is placed so as to be on the optical axis of the electron optical column 1.

  In order to observe the sample 13 with a scanning electron microscope, first, electrons are emitted from the electron gun 3 and the condenser lens 5, the astigmatism correction device 7, and the objective lens 8 are operated. Irradiate the line. Further, the scanning coil 6 is driven to deflect the electron beam and scan the surface of the sample 13.

  At the same time, the reflected electrons from the sample 13 are measured by the electron detector 31 for the scanning electron microscope, and the signal detected by the detector 31 and the signal for scanning the electron beam are displayed in synchronization with each other. Can be observed.

  In addition, in order to sample a small thin piece sample from the sample 13, an image of the sample 13 is observed with a scanning electron microscope, the focus is adjusted to the sample surface, and a portion of the sample to be sampled is determined. Next, the manipulator 30 is operated to bring the needle 35 close to the sample surface, and a necessary portion of the sample 13 is peeled off. In some cases, the sample stage 38 may be operated to cut a part of the sample. Finally, as shown in FIG. 4, the micro thin sample 34 is attached to the tip of the needle 35. The sample 13 after the fine thin sample 34 is peeled off is retreated to the sample introduction chamber 39 together with the sample stage 38.

  Next, an observation method of a minute thin piece sample with an electron microscope will be described with reference to FIG. When the micro thin sample 34 sampled at the tip of the needle 35 is observed with an electron microscope, the sample 34 must be irradiated with an electron beam under high vacuum in order to perform high resolution observation and transmission electron microscope observation. .

Therefore, first, the temperature control mechanism 40 of the manipulator 30 using a Peltier element cools the volatile substance containing the minute thin piece sample 34 to a temperature at which it does not evaporate under high vacuum. Thereafter, the gas introduction part 19 is closed to stop the gas flow, and the high vacuum evacuation of the lens chamber 9 and the sample chamber 2 of the electron optical column 1 is performed from the exhaust parts 17 and 18 (the vacuum of the lens chamber 9 and the sample chamber 2). The degree is, for example, about 10 ⁻³ Pa).

  Here, when performing electron microscope observation, an electron microscope image can be obtained simultaneously by the electron detector 31 for a scanning electron microscope and the electron detector 32 for a scanning transmission electron microscope. Specifically, as in the case of performing an electron microscope observation at the time of sampling of the minute thin piece sample 34, a finely focused electron beam is scanned on the surface of the minute thin piece sample 34, and secondary electrons emitted from the surface are scanned. Measurement is performed by the electron detector 31 for the scanning electron microscope, and at the same time, the electrons transmitted and scattered through the sample 34 are respectively measured in two areas of the electron detector 32 for the scanning transmission electron microscope. A secondary electron image, a bright-field image, and a dark-field image are obtained by displaying the signals measured by these two electron detectors 31 and 32 in synchronization with the signal for scanning the electron beam. I can do it. In addition, it is necessary to observe the acceleration voltage of the electron beam irradiated in order to obtain a scanning transmission electron microscope image with the acceleration voltage which an electron permeate | transmits a sample. As a result, it is possible to obtain information on the internal structure of the minute thin piece sample 34 from observation of the outer shape of the fine thin piece sample 34 from observation of the secondary electron image and observation of a bright field image and dark field image of the scanning transmission electron microscope.

  Here, secondary electrons are obtained by detecting secondary electrons with the electron detector 31, but this is purely for observing the external shape of the sample. The sample can be observed even if the backscattered electrons are detected.

  In this embodiment, as in the first and second embodiments, an arbitrary part of a volatile substance-containing sample such as an organic substance or a living organism is used as a minute thin piece sample that can be observed by a transmission electron microscope without being altered. Can be sampled easily. In addition, on the spot, it is possible to observe the external shape and the internal structure of the micro thin sample sampled by electron microscope observation in the sample chamber 2.

(Fourth embodiment)
FIG. 5 is a block diagram showing a fourth embodiment of the present invention. In the fourth embodiment, a manipulator 42 having an EDS (Energy Dispersive X-ray Spectrometer) detector 41 for detecting characteristic X-rays and a needle rotation and temperature control mechanism 43 is added to the configuration of the third embodiment. The three-dimensional structure of the micro thin sample 34 sampled at the tip of the needle by high-inclination electron microscope observation using a needle rotation mechanism can be observed.

  The basic configuration of this embodiment is the same as that of the third embodiment of FIG. 3, and an EDS detector 27 and a manipulator 42 other than the manipulator 30 are provided in the sample chamber 2. The manipulator 42 has a needle 35 at its tip, and a rotation and temperature control mechanism 43 comprising a rotation mechanism for rotating the needle 35 about its axis and a mechanism for controlling the temperature of the needle 35.

  Further, the needle 35 of the manipulator 42 is driven in the XYZ directions by the manipulator stage 36 and can be moved in the Φ direction by the guide rail 37. The needle 35 of the other manipulator 30 is keyed so that a sample of a small thin piece can be easily sampled. Since this manipulator 30 is used exclusively for the production of a minute thin piece sample, no temperature control mechanism is provided. The other configuration is the same as that of FIG. Since the key shape at the tip of the needle is very small, it is not shown in FIGS.

  The shape of the manipulator is not limited to the needle type. For example, a scissor type, a vacuum suction nozzle using working exhaust, or the like may be used. In addition, when the target sample is small, carbon nanotubes can be used as needles.

  Next, a sampling method for a micro thin sample according to the present embodiment will be described with reference to FIGS. The basic sampling method of the minute slice sample is the same as that of the third embodiment. First, the micro thin sample 34 is sampled at the tip of the needle 35 by the same operation as in the third embodiment using the manipulator 30 having the needle at the tip. Next, for observation with an electron microscope, sampling is performed by moving the micro thin sample 34 from the needle tip of the manipulator 30 to the needle tip portion of a manipulator 42 having a tip provided with a rotation and temperature control mechanism.

  Next, an electron microscope observation method for a minute thin piece sample will be described. First, in order to perform high-resolution observation and transmission electron microscope observation of a micro thin sample 34 sampled at the tip of a manipulator 42 having a needle equipped with a rotation and temperature control mechanism at the tip, the same as in the third embodiment. It is necessary to irradiate the minute sample piece 28 with an electron beam under vacuum.

Accordingly, the micro thin sample 34 is cooled by the temperature control mechanism 43 of the needle in the manipulator 42 as in the third embodiment, and further, high vacuum evacuation of the lens chamber 9 and the sample chamber 2 of the electron optical column 1 is performed. (For example, the degree of vacuum is about 10 ⁻³ Pa). Thereafter, in order to observe the micro thin sample 34 from various directions with an electron microscope, the needle 35 of the manipulator 42 is moved along the guide rail 37, and the rotation axis of the needle 35 of the manipulator 42 is changed to an electron beam as shown in FIG. Make orthogonal.

  When performing electron microscope observation, in the same manner as in the third embodiment, the electron detector 31 for the scanning electron microscope detects secondary electrons from the sample, and a signal for scanning the electron beam. Are synchronized to obtain a secondary electron image. In this case as well, the sample may be displayed by detecting reflected electrons instead of secondary electrons. At the same time, as in the third embodiment, the electron detector 32 for the scanning transmission electron microscope detects the electrons transmitted and scattered through the sample, and synchronizes the signal for scanning the electron beam with the bright field image. And a dark field image is obtained.

  Further, in the microscopic observation, the microscopic thin piece sample 34 at the tip of the needle is rotated at various angles as shown in FIG. 6 by the rotation of the needle of the manipulator 42 and the temperature control mechanism 43, and the electron microscopic observation is performed at the same time. .

  Therefore, secondary electron images, bright field images, and dark field images from various directions are obtained, and by combining this with image processing by a computer (not shown), the three-dimensional external structure and internal structure of the micro thin sample 34 are obtained. Information can be obtained. Furthermore, by combining elemental mapping by the EDS detector 41 with this information, it is possible to construct a three-dimensional distribution of the composition.

  In the present embodiment, an arbitrary part of a volatile substance-containing sample such as an organic substance or a living organism can be easily sampled as a small thin piece having a size that can be observed with a transmission electron microscope without being altered. In addition, it is possible to perform a three-dimensional appearance observation, an internal structure observation, and a composition analysis of a minute thin piece sample by high-inclination electron microscope observation on the spot.

It is a block diagram which shows the 1st Embodiment of this invention. It is a block diagram which shows the 2nd Embodiment of this invention. It is a block diagram which shows the 3rd Embodiment of this invention. It is a figure explaining the electron microscope observation method of the organic substance micro thin sample of the 3rd Embodiment of this invention. It is a block diagram which shows the 4th Embodiment of this invention. It is a figure explaining the electron microscope observation method of the organic micro thin piece sample of the 4th Embodiment of this invention.

Explanation of symbols

1 Electro-optical column 2 Sample chamber (sample chamber)
DESCRIPTION OF SYMBOLS 3 Electron gun 4 Electron gun chamber 5 Condenser lens 6 Scanning coil 7 Astigmatism correction device 8 Objective lens 9 Lens chamber 10, 11 Orifice 12 Manipulator 13 Sample containing volatile substance 14 Sample holder 15 Sample holder 16 for transmission electron microscope DESCRIPTION OF SYMBOLS 18 Exhaust part 19 Gas introduction part 20, 21 Preliminary exhaust chamber 22 Backscattered electron detector 30, 42 Manipulator 31 Electron detector for scanning electron microscope 32 Electron detector for scanning transmission electron microscope 34 Micro flake sample 35 Needle 36 Manipulator Stage (XYZ)
37 Needle guide rail (Φ)
38 Sample stage (XYZΘ)
39 Sample introduction chamber 40 Needle temperature control mechanism 41 EDS detector 43 Needle rotation and temperature control mechanism

Claims (11)

A sample chamber capable of controlling the atmosphere, a first sample holder for introducing a volatile substance-containing sample into the sample chamber, a scanning electron microscope for observing the sample held in the sample holder, and the first And a second sample holder for holding a sample sampled by the manipulator and taking it out of the sample chamber. Micro thin piece production equipment. The micromanufacturing device according to claim 1, wherein the manipulator can be operated from the outside of the sample chamber and has a needle for sampling a sample at a tip. The scanning electron microscope includes an electron optical barrel including an electron gun chamber having an electron gun and a lens chamber having a lens, and the two chambers are adjusted to a predetermined different degree of vacuum by individual exhaust systems. The apparatus for producing a micro thin piece according to claim 1. 2. The apparatus for producing a micro thin piece according to claim 1, wherein at the time of sampling, the sample chamber is evacuated to a degree of vacuum so that the volatile substance-containing sample is not altered. 2. The micro thin piece manufacturing apparatus according to claim 1, wherein the scanning electron microscope displays an image of a sample surface by synchronizing a signal for scanning an electron beam and a sample current flowing in the sample. . The scanning electron microscope includes a detector that detects the current of the sample, the needle, and the second sample holder, and synchronizes the current detected by the detector with a signal for scanning the electron beam. 2. An apparatus according to claim 1, wherein an image obtained by combining the images of the sample, the needle, and the second sample holder is displayed. The scanning electron microscope includes a sample held by the first sample holder, a needle, a reflected electron detector for detecting reflected electrons from the second sample holder, and a signal detected by the reflected electron detector; The apparatus for producing a minute slice according to claim 1, wherein an image of the sample surface, the needle, and the second sample holder is displayed by synchronizing with a signal for scanning the electron beam. 2. The apparatus according to claim 1, wherein the manipulator is temperature-controllable, and further includes an electron detector for observing the sampled sample by the scanning electron microscope. apparatus. An electron detector for observing a sample sampled in the sample chamber with the scanning electron microscope, an electron detector for observing a scanning electron microscope image, and a bright field image and a dark field of a scanning transmission electron microscope image 9. The apparatus according to claim 8, further comprising an electron detector having one or a plurality of detection regions for image observation. The temperature-controllable manipulator is provided with one or a plurality of manipulators that have a rotating mechanism for rotating a needle shaft to a manipulator that holds a sampled sample, and three-dimensional morphology observation of a micro thin sample by rotating the needle shaft The apparatus for producing a micro thin piece according to claim 8, wherein: At the time of observing the sampled sample, the temperature control mechanism of the manipulator cools the sampled sample sample and the electron optical column to a higher vacuum by cooling to a temperature at which the volatile substance containing the sampled sample does not volatilize. The apparatus for producing a thin flake according to claim 8, wherein the apparatus is evacuated every time.

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