KR101529145B1 - Work stationcomprising sample holder for cryogenic electron microscopy for correlative imaging detection apparatus in combination of optical microscopy and electron microscopy, correlative imaging detection including said work station, imaging detection method and imaging system by using said work station - Google Patents

Abstract

The present invention relates to a work station for cooperative measurement between an optical microscope and an electron microscope, which is formed on an upper surface of a work station (10) for placing a specimen grid on an optical microscope, A grid accommodating groove 340 for positioning the specimen grid in a space separated by the cooling water accommodating portion 310 and the accommodating portion bulkhead 323, And a grid accommodating portion 150 having a through hole 156 extending to the outside from one side of the grid accommodating groove 340. The liquid accommodating portion 150 is provided with a liquid The seating head 380 for seating the specimen grid is formed at the tip of the extension pipe 180 to which the nitrogen is supplied so that the seating head 380 is inserted into the through hole 156, And a specimen holder (14) detachably attached to the specimen holder (14) so as to position the specimen holder (14). The workstation (1) includes a cryogenic electron microscope specimen holder And to an imaging detection method and an imaging system using the same.

Description

TECHNICAL FIELD The present invention relates to a workstation including a cryo-sample holder for cryo-imaging detection of an optical microscope and an electron microscope, an associated imaging detection apparatus including the same, and an imaging detection method and an imaging system using the same. a correlated imaging detection apparatus in combination with optical microscopy and electron microscopy, an imaging detection method and imaging system,

TECHNICAL FIELD The present invention relates to a workstation for cooperative measurement between an optical microscope (light microscope) and an electron microscope (Electron Microscope), a cooperative measurement device including the same, and a measurement method and a measurement system using the same.

2. Description of the Related Art In general, light microscopes and electron microscopes are widely used as devices for observing biological samples in medical and life science fields.

An optical microscope is a microscope designed to examine the position and characteristics of a specific element or element by injecting a fluorescent dye that emits fluorescence under ultraviolet light close to visible rays. The fluorescent dye used here is characterized by absorbing ultraviolet light of short wavelength and emitting energy of long wavelength. For example, an optical microscope is used to immunologically test an antibody that can react with a specific antigen when the immunoassay is performed in a hospital. Such an optical microscope can be used for a sample which has fluorescence itself or can be adsorbed to a fluorescent substance. Therefore, the optical microscope is used for identification of an infection pathway of a bacteria or a virus, And the detection of biological phenomena such as the expression of specific proteins according to the present invention and the detection / inspection of biological samples in cells.

On the other hand, an electron microscope is an apparatus in which an electron beam is used in place of visible rays used in an optical microscope and an electron lens is used instead of a glass lens to form an enlarged image of an object . Such an electron microscope is a scanning electron microscope (SEM) that scans a surface of a sample placed in a vacuum with fine electron beams to display an enlarged image, and accelerates electrons emitted from the filament to transmit an electron beam exiting the hole of the anode to the specimen And transmission electron microscopy (TEM) in which an image is obtained and the image is enlarged by an electron lens.

In recent years, techniques for accurately observing the surface and internal shape of micro-materials due to the industrialization of ultrafine technology in the industrial field have been gradually developed. With the introduction of such an electron microscope, the technology in medical biotechnology has been rapidly developed . Since the electron microscope has high resolution, it can observe the object at a higher magnification than the optical microscope. In addition, it can identify the relationship between the microstructure of the object and the physical properties such as hardness and reflectivity. It has the advantage of being able to analyze the size, types of elements and compounds and their relative amounts.

However, although the electron microscope can observe a state within a very small area at a high magnification, it has a disadvantage that it is not suitable for observing a feature of a large area. Therefore, in recent years, there has been a tendency to use an optical microscope and an electron microscope to observe living cells, to extract necessary signals through an optical microscope, and then to observe the extracted regions intensively through an electron microscope.

Conventionally, in order to observe a living body sample in conjunction with an optical microscope and an electron microscope, for example, a living body sample is made to be clouded with a liquid and then placed on a support film which has been subjected to a predetermined treatment with a pipette. Water is removed by a filter paper or the like, And a rapid cooling process is performed so that ice crystals do not occur. Thereafter, a rapidly cooled biological sample grid is transported by an optical microscope and then observed with an electron microscope once again.

Therefore, conventionally, a process of rapidly cooling the biological sample through a separate device and then transporting the biological sample through an optical microscope, and a manual operation through a tool such as tweezers in the process of transporting the biological sample observed by the optical microscope with an electron microscope I can not help it. Therefore, conventionally, damage or damage of the sample grid occurs in each process of carrying the sample grid in which the signal is extracted by the preprocessing process and the optical microscope. In addition, since the sample grid pretreated by the rapid cooling has to keep the temperature constant during the final transportation by the electron microscope, the sample transporting operation is very troublesome, troublesome, and time consuming.

As a technique for solving such a problem, Korean Patent Registration No. 10-1396420 filed by the applicant of the present invention registers a pretreatment process in which a sample grid is immersed in liquid nitrogen to cool the sample grid, and a preprocessed sample grid is transported to an optical microscope And a series of processes for observing and observing a specimen are continuously performed in a single specimen preparation apparatus. 10-1396420 discloses a method of injecting liquid nitrogen through an injection port 2 formed in a main body 1 of a specimen preparation apparatus A as shown in FIG. The sample grid is taken out of the sample grid carrying container placed in the sample storage part 5 of the sample loading part and mounted on the sample loading part 4. The sample preparation device A equipped with the sample grid is mounted on an optical microscope A technique is described that can be performed.

However, in the Korean Registered No. 10-1396420, the sample grid can be directly mounted with an optical microscope after the pretreatment, but the introduction into the electron microscope can still be accomplished by manually transporting the grid. In particular, such an electron microscope has a complicated measuring environment and a mounting environment of the sample grid, so that the transportation of the sample grid is very difficult and a lot of time .

Therefore, it is urgently required to solve the above-mentioned problems of the prior art and to introduce a technology for preliminarily processing a sample grid and continuously inputting not only an optical microscope but also an electron microscope.

Domestic Registration Bulletin 10-1396420 (May 20, 2014)

In view of the above, the present invention, which has been developed in view of the above, enables a pretreatment process of a specimen grid, a measurement through an optical microscope and an electron microscope to proceed through a single apparatus, And it is an object of the present invention to provide a linkage measuring apparatus for linkage measurement of an optical microscope and an electron microscope, which enables the cooling state to be maintained continuously, and a measurement method using the same.

In a workstation in which a pretreatment process for cooling a specimen grid according to the present invention is performed, and a specimen grid is inserted into a specimen holder, and observation of images in an optical microscope and an electron microscope is continuously performed without carrying the specimen grid,

A coolant receiving portion in which the liquid nitrogen is received, and a coolant receiving portion formed separately from the coolant receiving portion, and the sample grid is positioned And a grid accommodating portion formed in the grid accommodating groove to include a temperature sensor for measuring a temperature of the specimen grid and a through hole formed at one side of the through hole.

Further, in the coupled-type imaging detection apparatus of an optical microscope and an electron microscope according to the present invention,

A cooler in which the work station and the liquid nitrogen are stored for performing a pretreatment process for cooling the specimen grid, and an extension pipe extending to one side of the cooler, and is connected to the work station during the preprocessing process, And a specimen holder which is detached from the workstation and inserted into the measurement part of the electron microscope for connection measurement.

Further, a linked imaging system for linked measurement of an optical microscope and an electron microscope according to the present invention,

A work station for performing a pretreatment process for cooling a specimen grid, a cooler for storing liquid nitrogen, and an extension pipe extending to one side of the cooler, wherein the work station is connected to the work station during a preprocessing process, An optical microscope for measuring a signal for confirming an analyte on a specimen grid preprocessed by the workstation, and an optical microscope for measuring a signal for confirming an analyte on the specimen grid preprocessed by the workstation, And an electron microscope for analyzing the signal measured by an optical microscope.

Finally, an imaging detection method in which an optical microscope and an electron microscope are combined according to the present invention,

Placing a work station having a grid receiving portion formed at a spaced apart position below the optical microscope and including a through hole formed at one side of the specimen grid and capable of placing a specimen grid thereon, Placing the specimen grid within the grid housing, cooling the specimen grid, placing the cooled specimen grid in the specimen holder, forming a signal on the cooled specimen grid using an optical microscope, And separating the specimen mounting holder from the workstation after forming the signal.

According to the present invention, the pre-process for cooling the specimen grid and the measurement process through the optical microscope are performed in a single coherence measuring apparatus, and the measurement is made through an optical microscope, followed by an electron microscope By making it possible to load the specimen without moving the specimen grid for input, it is possible to reduce the time required to move the specimen grid and prevent damage to the specimen grid.

Also, according to the present invention, it is possible to continuously maintain the freeze state of the specimen grid during the movement of the specimen grid after the measurement through the optical microscope, thereby further improving the accuracy of the measurement through the electron microscope.

1 is a perspective view of a work station and a linkage measuring apparatus including the work station according to the first embodiment of the present invention;
FIG. 2 is a side view of the workstation shown in FIG. 1 and a linkage measuring device including the same.
3 is an enlarged perspective view of the grid receiving portion of the work station shown in Fig.
Fig. 4 is a plan view and side view of the grid receiving portion shown in Fig. 3; Fig.
5 is a perspective view of a work station and a linkage measuring apparatus including the work station according to the second embodiment of the present invention.
6 is a side view of the workstation shown in Fig.
7 is an enlarged perspective view of the grid receiving portion of the work station shown in Fig. 5;
8 is a plan view and side view of the grid receiving portion shown in Fig.
FIG. 9 is an enlarged perspective view of a measurement lens holder according to the first and second embodiments of the present invention; FIG.
10 is a side view of a specimen mounting holder according to the first and second embodiments of the present invention.
11 is a perspective view of a work station and a linkage measuring apparatus including the work station according to the first embodiment of the present invention.
12 is a perspective view showing a conventional technique.

Hereinafter, exemplary embodiments of the present invention will be described in detail with reference to the accompanying drawings, which illustrate exemplary embodiments of the present invention. The present invention is not limited to these embodiments.

FIG. 1 is a perspective view of a work station and a linkage measuring apparatus including the work station according to an embodiment of the present invention, and FIG. 2 is a side view of the linkage measuring apparatus including the work station shown in FIG.

Referring to FIGS. 1 and 2, the cooperative measurement apparatus according to the present embodiment includes a work station 10 for performing a preprocessing process for cooling a specimen grid, and a pre- And a specimen mounting holder 14 which is detached from the work station 10 and inserted into the measurement part of the electron microscope for connection measurement after the process is performed.

The work station 10 includes a box unit 120 in which a grid housing unit 150 is formed and an external terminal unit provided at a lower portion of the box unit 120 and connected to an external measuring instrument or control device, An upper plate 110 which covers the upper portion of the box portion 120 and has an opening for exposing the grid accommodating portion 150 upwardly and an upper plate 110 vertically supporting the upper plate 110, A base plate 140 supporting the lower end of the vertical support 123 and a horizontal support 142 formed on one side of the base plate 140 and supporting the work station 10 horizontally As shown in FIG.

The first handle 121 and the second handle 141 are provided on the upper portion of the box portion 120 and the base plate 140, respectively. The second handle 141 is formed in a structure capable of rotating in a vertical direction and a horizontal direction. For example, the second handle 141 is rotatable in vertical and horizontal directions with a hinge formed on the handle. Thereby allowing easy separation when detaching from the workstation 10 for insertion of the specimen mounting holder into the electron microscope.

Further, the box portion 120 is formed of a thermally conductive material including copper on the inner side, and the outer side is formed of the hardest acetal material of the resin type. In particular, a heat insulating layer formed of a heat insulating material including styrofoam is formed in a multilayer structure between the inside and the outside.

The grid receiving part 150 is formed so that a specimen grid to be measured can be received.

The grid accommodating unit 150 includes an inflow hole 124 into which an inflow pipe 124 for supplying liquid nitrogen for cooling the specimen grid from the liquid nitrogen tank (not shown) provided outside the work station 10 is inserted And a discharge hole 325 into which a discharge pipe 125 for discharging the liquid nitrogen supplied to the inside is inserted.

Here, valves (not shown) are provided in the inlet pipe 124 and the outlet pipe 125, respectively, so that the inflow and outflow of liquid nitrogen can be controlled. This allows the workstation 10 to be easily moved by an electron microscope.

The grid housing unit 150 includes a housing cover 152 that covers or opens the upper portion of the housing cover 152. A handle 154 is formed at an upper portion of the housing cover 152 so as to be easily hand- do.

For reference, the receptacle cover 152 may be formed of a transparent material such as acryl, glass, or synthetic resin, but may be formed of various materials. Preferably, the receptacle cover 152 is formed of a material containing acrylic to facilitate observation.

In addition, the receptacle cover 152 may be formed with an opening 155 in the form of an elongated hole for inserting a work tool into the grid receiving portion 150 while covering the upper portion of the grid receiving portion 150 have.

In addition, according to the present invention, a work station 10 for performing a pretreatment process for cooling a specimen grid, a cooler 170 for storing liquid nitrogen, and an extension pipe (not shown) extending to one side of the cooler 170 180), which is connected to the work station (10) during the preprocessing process, and which is detached from the workstation (10) after the preprocessing process is performed and inserted into the measurement part of the electron microscope (14), and a linkage measuring device for linkage measurement of the electron microscope are provided.

In the present embodiment, the above-mentioned work station can be applied to the work station, and a description thereof will be omitted below.

The specimen mounting holder 14 is detachably mounted on one side of the work station 10 and the distal end of the elongated tube 180 extending to one side is inserted into the side of the work station 10, (Not shown).

At this time, the specimen mounting holder 14 having various lengths seats the cooler 170 on the base plate 140, supports the lower part of the cooler 170, slides in a direction separating from the grid receiving part 150 And a sliding guide 190 having a stopper for restricting insertion in a direction of insertion into the grid receiving part 150. [ Further, the sliding guide 190 is slid back and forth so that the sample holder 14 can be easily moved.

Fig. 3 is an enlarged perspective view of the grid receiving portion of the linked measuring device shown in Fig. 1, and Fig. 4 is a plan view and a side view of the linked measuring device shown in Fig.

3 and 4, the grid receiving part 150 forms an inner space opened toward the upper part of the work station 10, and is formed in the box part 120, And is exposed upward through the opening.

The grid accommodating portion 150 includes a coolant accommodating portion 310 for accommodating the liquid nitrogen supplied therein and a grid accommodating groove 340 for forming a space in which the specimen grid is separated from the coolant accommodating portion 310 And a through hole 156 penetrating to the outside of the work station 10 is formed on one side of the inside.

The grid accommodating portion 150 further includes an inflow hole 324 and an exhaust hole 325 at the other side of the inside. The inflow hole 324 introduces liquid nitrogen, and the discharge hole 325 discharges liquid nitrogen. In particular, the discharge hole 325 is formed at a position higher than the inflow hole 324 so that the liquid nitrogen can be discharged while maintaining a constant water level.

Shaped inlet barrier ribs 312 are formed on the inner side of the grid accommodating portion 150 to prevent the liquid nitrogen flowing from the inlet holes 324 from splashing. The coolant accommodating portion 310 and the grid accommodating groove 340 are separated from each other by the accommodating portion barrier rib 323 so that the liquid nitrogen contained in the coolant accommodating portion 310 is not introduced into the grid accommodating groove 340.

Here, the accommodating portion bulkhead 323 includes copper and is designed to be slightly lower than the height of the entire box portion. This design closes the receptacle cover 152 during image observation and causes the cold air of liquid nitrogen to pass in the direction of the grid receiving groove 340 where the specimen grid where the frozen grid is located in the grid receiving portion 150, Do not melt without.

The grid accommodating groove 340 is formed to accommodate the specimen grid therein. The lower portion of the grid accommodating groove 340 is provided with a lens groove (not shown) through which a measurement lens for measuring the specimen grid can be inserted through the optical microscope (342) is formed at the bottom.

The workstation according to the present invention is for measuring a specimen grid by connecting a specimen grid with an optical microscope and an electron microscope. For example, when observing a specimen of a living body cell, the position of a signal observed through an optical microscope is prevented from being changed It is preferred, but not limited, to use liquid nitrogen for rapid freezing of the specimen grid.

The grid accommodating part 150 includes a cooling groove 320 for inserting a grid holder (not shown) for storing a plurality of the specimen grids into a lower portion of one side of the coolant accommodating part 310 to maintain the liquid holder in a locked state .

Further, a holder seating groove 330 is formed to take out the grid holder from the cooling groove 320 and to seat it on one side of the grid receiving groove 340.

For reference, the grid holder may be formed in a circular shape, and the cooling groove 320 and the holder seating groove 330 may be formed in a circular shape so as to be inserted through the grid holder.

At this time, the cooling grooves 320 and the holder seating grooves 330 may be formed at one side of the grid holder so as to form protrusions 321 and 331, respectively, each having a concavo-convex structure to prevent the grid holder from moving. In particular, the holder seating groove 330 can reliably retrieve the specimen grid from the grid holder.

In addition, a temperature sensor 343 for measuring the temperature can be formed inside the grid accommodation hole, and the temperature can be adjusted by the user by sensing the internal temperature cooled by the coolant.

In detail, the temperature measured by the temperature sensor 343 serves as a basis for controlling an external control device connected through the external terminal unit 122, and the external control device controls the flow of liquid nitrogen to control the temperature of the sample grid .

FIG. 5 is a perspective view of a work station and a linkage measuring apparatus including the work station according to a second embodiment of the present invention, and FIG. 6 is a side view of the linkage measuring apparatus including the work station shown in FIG.

5 and 6, the workstation according to the second embodiment is the same as the workstation according to the first embodiment in basic configuration and operation, but the workstation according to the second embodiment is more portable.

The work station 10 is configured such that the inlet pipe 124 and the inlet hole 324 from which liquid nitrogen is supplied from the outside are removed so as to be easily moved by an electron microscope but the outlet 125 and the outlet hole 325 are connected to each other through the liquid nitrogen overflow and to prevent overflow.

The housing cover 152 is formed with a first inflow hole 156 and a second inflow hole 157. The first inflow hole 156 is inserted into the first inflow pipe 360, (157) is inserted into the second inflow pipe (370). Details of the first inflow pipe 360 and the second inflow pipe 370 will be described later.

Configurations other than the above-described configuration are the same as those of the workstation according to the first embodiment.

Fig. 7 is an enlarged perspective view of the grid receiving portion of the work station shown in Fig. 5, and Fig. 8 is a plan view and a side view of the grid receiving portion shown in Fig.

Referring to FIGS. 7 and 8 together, the grid receiving portion 150 according to the second embodiment has the inlet hole 324 removed, and only the outlet hole 325 is retained.

The grid accommodating part 150 receives the liquid nitrogen contained in the portable cooling part 350 through the first inflow pipe 360 and the second inflow pipe 370. The portable cooling portion 350 may be formed in a funnel or a container shape. The first inlet pipe 360 introduces liquid nitrogen into the grid receiving groove 340 and the second inlet pipe 370 receives liquid nitrogen into the coolant receiving portion 310.

The first inflow pipe 360 may be inserted into the first inflow hole 156 to allow liquid nitrogen to flow into the grid accommodating groove 340 and the second inflow pipe 370 may be connected to the second inflow hole 157, So that the liquid nitrogen can be introduced into the coolant accommodating portion 310.

Or the first inlet pipe 360 and the second inlet pipe 370 may flow liquid nitrogen through the opening portion 155 of the receiving portion cover 152 shown in the first embodiment.

At this time, the first inflow pipe 360 connected to the portable cooling unit 350 is basically formed at a higher position than the second inflow pipe 370. Accordingly, the first inflow pipe 360 can stop the inflow of liquid nitrogen more quickly than the second inflow pipe 370. This allows the liquid nitrogen to flow into the coolant accommodating portion 310, 340 can stop the flow of liquid nitrogen and facilitate image measurement.

Further, the first inlet pipe 360 and the second inlet pipe 370 may further include valves to control the inflow amount of liquid nitrogen.

The configurations other than the above-described configuration are the same as the grid receiving portion 150 according to the first embodiment.

9 is an enlarged perspective view of a measurement lens holder according to the first and second embodiments of the present invention.

Referring to FIG. 9, the measurement lens holder 400 is provided to prevent freezing of the measurement lens. The measurement lens holder 400 includes a holder body 410, a gas injection unit 420, and a buffer 430.

The holder main body 410 is inserted with a measuring lens and the gas injecting part 420 is formed on one side of the holder main body 410 in order to inject an inert gas such as nitrogen gas to prevent the measurement lens from freezing . The buffer 430 is installed to prevent shock absorption and vibration of the measurement lens inserted in the holder body 410. That is, the buffer 430 protects the measurement lens from external impact.

Therefore, the measurement lens holder 400 secures the problem that when the sample cooled by the liquid nitrogen and the measurement lens come close to each other, the measurement lens is frozen due to the temperature difference, and the glaze is generated and the accurate image can not be obtained.

Fig. 10 shows a side view of the specimen mounting holder shown in the first and second embodiments of the present invention.

5, the specimen mounting holder 14 includes a cooler 170 capable of storing liquid nitrogen therein, an extension tube 180 extending to one side from the cooler 170 to supply liquid nitrogen, And a seating head 380 provided with a grid seating groove 382 for seating the specimen grid.

The cooler 170 is desirably formed in a small size so that it can be easily carried and moved by hand.

The liquid nitrogen stored in the cooler 170 is supplied to the seating head 380 through the extension tube 180 to supply liquid nitrogen to the specimen grid seated in the seating head 380.

The specimen mounting holder 14 is constructed so that the positioning head 380 formed at the tip of the extension tube 180 can be inserted into a conventional measurement unit of the electron microscope to position the specimen grid.

At this time, the extension pipe 180 of the specimen mounting holder 14 is formed with a plurality of steps at which the diameter is reduced at the tip end side so that the extension pipe 180 can be accurately stopped at the inserted position, May be formed to accurately position the specimen grid within the station 10.

Here, the extension pipe 180 is formed to extend to one side of the box part 120 in which the coolant is stored, the inner side is formed of a heat insulating material such as styrofoam, and the outer side is formed of the hardest acetal material among resin types.

The extension pipe 180 may be provided with a toothed wheel so as to adjust its thickness and length according to the specimen mounting holder 14 having various thicknesses and lengths or may be provided with an extension pipe 180 depending on the size of the specimen mounting holder 14 And can be formed in a detachable structure according to the size.

Hereinafter, the operation of the work station and the linkage measuring apparatus including the work station according to the present invention will be described in detail.

According to the present invention, in linking the optical microscope with the measurement of the electron microscope, the grid holder 150 of the work station 10 for measurement of the optical microscope and the specimen mounting holder 150 detachably attachable to the measuring portion of the electron microscope, respectively, Temperature specimen grid is mounted on the grid mounting groove 382, which is the end portion of the grid 14, so that measurement can be performed.

According to the present invention, before placing the specimen grid on the optical microscope, the positioning head 380 of the specimen holder 14 is placed in the grid receiving groove 340 of the grid receiver 150 in the work station 10 . After the grid holder 150 is filled with liquid nitrogen and the grid holder with the cryogenic specimen stored therein is first placed in the receiving part 320 and the temperature of the grid receiving groove 340 is stabilized below -185 degrees, And is placed in the holder receiving groove 330 of the grid receiving part 150 and the specimen grid is placed on the seating head 380 of the specimen mounting holder 14 positioned adjacent thereto using a mechanism such as a tweezer ), And the measurement is carried out with a lens (not shown) located under the optical microscope

When the signal is measured through the optical microscope, the specimen holder 14 is detached from the work station 10 to separate the seating head 380, on which the specimen grid is seated, from the grid receiver 150 and then to the inside of the electron microscope And the imaging measurement based on the signal of the optical microscope is carried out.

At this time, from the time of separating the specimen grid measured through the optical microscope from the work station 10, the liquid nitrogen by the cooler 170 of the specimen holder 14 continuously flows into the specimen holder 14 through the electron microscope until the measurement is completed So that the state of the specimen grid according to the signal measured through the optical microscope can be accurately maintained at a cryogenic temperature.

Therefore, according to the present invention, the pre-processing for cooling the specimen grid and the measurement through the optical microscope are performed in a single linked measuring apparatus, and it is necessary to transfer the specimen grid after measurement through the optical microscope It can be put into the measurement part of the electron microscope without any modification.

Therefore, it is possible to eliminate the inconvenience of manually moving the specimen grid for the electron microscope measurement and to maintain the freezing of the specimen grid during the transfer to the electron microscope, thereby improving the accuracy of the measurement through the electron microscope .

In addition, it is possible to prevent damage and breakage in the process of moving the specimen grid by hand.

11 is a perspective view of a work station and a linkage measuring apparatus including the work station according to the first embodiment of the present invention.

Referring to FIG. 11, a height adjusting device 510 for supporting the lower surface of the base plate 140 of the linkage measuring apparatus according to the previous embodiment and adjusting the height of the linkage measuring apparatus is further included.

The height adjusting device 510 includes a ground plate 520 supporting the floor of the ground, a folding member 540 forming up and down driving strokes by a plurality of links between the ground plate 520 and the base plate 140, And a driving lever 550 for driving the folding member 540 up and down by inserting an operating shaft 530 formed with threads into the folding member 540.

According to such a configuration, the specimen holder 14 can be moved integrally up and down in a state in which the specimen holder 14 is inserted into the work station 10 positioned in the optical microscope, and the accuracy of measurement through the optical microscope can be increased. So that it is possible to perform the measurement more efficiently and easily.

Hereinafter, a measurement method in which an optical microscope and an electron microscope are combined according to another aspect of the present invention will be described.

For reference, the measurement method according to the present embodiment can be performed using the above-described work station and the linked measurement apparatus including the same.

Hereinafter, when referring to the configuration of the linkage measuring apparatus, constituent elements having the same functions as those of the above-mentioned reference numerals are denoted by the same reference numerals, and descriptions overlapping with those of the above- To avoid this, a brief explanation will be given.

According to this embodiment, the specimen mounting holder 14 is inserted into one side of the grid accommodating portion 150 for positioning the specimen grid in the optical microscope, and the specimen assembly holder 14 is inserted into the grid accommodating groove 340 of the grid accommodating portion 150, A first step of positioning the seating head 380 of the specimen mounting holder 14 on which the grid is mounted and a second step of placing the grid holder in which the cryogenic specimen is stored after filling the grid housing part 150 with liquid nitrogen into the accommodating part 320 The grill holder is lifted from the cooling water accommodating portion 320 and placed in the holder seating groove 330 of the grid accommodating portion 150 A second step of mounting a cryogenic grid held in a grid holder to a grid seating groove 382 serving as a sample inserting portion of the seating head 380 of the grid receiving portion 150 to measure a signal through the optical microscope Step 3 and above By separating the mounting holder 14 from the grid receiving portion it may be made of a fourth step of measuring the input and the mounting head 380 by an electron microscope.

In the third step, the specimen grid cooled by the grid holder is picked up by the tweezers and transferred to the seating head 380.

At this time, the grid holder is raised from the cooling water receiving portion 310 and can be easily moved to the grid receiving groove 340 positioned adjacent to the holder receiving groove 330 of the grid receiving portion 150.

Even after the grid holder is lifted from the cooling water accommodating portion 310, the liquid is continuously influenced by the nitrogen in the grid accommodating portion 150, so that the frozen state can be continuously maintained.

In the fourth step, the sample holder 14 is detached from the grid receiving part 150, and then the liquid by the cooler 170 provided in the sample holder 14 until the measurement by the electron microscope is completed Nitrogen is supplied to the specimen grid to maintain cooling.

As described above, according to the measurement method in which the optical microscope and the electron microscope are combined in accordance with the present invention, it is possible to rapidly inject the specimen through the optical microscope after cooling in the process of pretreatment of the specimen grid through the optical microscope, It is possible to keep the freeze state of the specimen grid constant.

In addition, since it is not necessary to transfer the specimen grid to an electronic microscope after measurement through an optical microscope, it is possible to easily and easily transfer the specimen grid by one device, Possible breakage can be prevented and continuous supply of liquid nitrogen by the cooler is possible.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments. It is to be understood that various changes and modifications may be made without departing from the scope of the appended claims.

10: Workstation 14: Specimen holder
110: upper plate 120: box part
121: first handle 122: external terminal portion
123: vertical support 124: inlet pipe
125: outlet pipe 140: base plate
141: second handle 142: horizontal support
150: Grid receiving part 152: Receiving part cover
154: Cover handle 155:
156: first inlet hole 157: second inlet hole
158: Through hole 170: Cooler
180: extension pipe 190: sliding guide
310: coolant receiving portion 312: inlet partition wall
320: cooling groove 323: accommodation partition wall
324: Inflow hole 325: Discharge hole
330: holder seating groove 340: grid receiving groove
342: lens groove 343: temperature sensor
350: portable cooling unit 360: first inlet pipe
370: second inlet pipe 380: seat head
382: grid seating groove 400: measuring lens holder
410: holder body 420: gas injection part
430: Buffer 510: height adjuster
520: ground plate 530: operating shaft
540: folding member 550: drive lever

Claims (29)

A workstation in which a preprocessing step of cooling a specimen grid and inserting a specimen grid into a specimen holder is performed continuously without image carrying of the specimen grid by optical microscopy and electron microscopy,
An inlet through which liquid nitrogen is supplied from the outside of the work station and an outlet through which the supplied liquid nitrogen is discharged;
A coolant receiving portion in which the liquid nitrogen is received;
A grid receiving groove formed separately from the coolant receiving portion and on which the specimen grid can be placed;
A temperature sensor formed in the grid receiving groove and measuring a temperature of the specimen grid;
A grid receiving portion including a through hole formed at one side to penetrate to the outside;
A box portion in which the grid accommodating portion is formed;
An upper plate covering an upper portion of the box portion and having an opening exposing at least a part of the grid accommodating portion;
A support for vertically supporting the upper plate; And
And a base plate for supporting a lower end of the support portion. The work station for cooperative measurement of an optical microscope and an electron microscope.
The method according to claim 1,
And a receptacle cover for opening or closing an upper portion of the grid accommodating portion. The work station for cooperative measurement of an optical microscope and an electron microscope.
3. The method of claim 2,
The receiving portion cover
And an opening portion in the form of an elongated hole for inserting a working tool into the grid accommodating portion in a state of covering an upper portion of the grid accommodating portion.
The method according to claim 1,
Wherein:
And an inflow hole formed on a side of the discharge hole, the inflow hole being formed at a lower height than the discharge hole and allowing the liquid nitrogen to flow in from the outside, and a work station for measuring the connection between the optical microscope and the electron microscope.
5. The method of claim 4,
Shaped inlet barrier rib is formed on the inside of the grid receiving part to prevent the liquid nitrogen flowing from the inlet hole from being splashed. The work station for the linkage measurement of the optical microscope and the electron microscope.
3. The method of claim 2,
The receiving portion cover is formed with two inflow holes,
Wherein:
The two inflow holes;
A first inlet pipe inserted into one of the two inlet holes to introduce liquid nitrogen into the grid receiving groove; And
And a second inlet pipe inserted into the other one of the first inlet pipe and the second inlet pipe to introduce liquid nitrogen into the coolant receiving portion.
The method of claim 3,
Wherein:
A first inlet pipe for introducing liquid nitrogen into the grid receiving groove; And
And a second inflow pipe for introducing liquid nitrogen into the coolant accommodating portion,
The first inlet pipe and the second inlet pipe are connected to each other,
And passing through the opening. The work station for the cooperative measurement of an optical microscope and an electron microscope.
8. The method of claim 7,
The first inlet pipe and the second inlet pipe are connected to each other,
A liquid cooling unit connected to the portable cooling unit to supply the liquid nitrogen while carrying the liquid nitrogen,
And the first inlet pipe is formed at a position higher than the second inlet pipe. The work station for the connection measurement of the optical microscope and the electron microscope.
The method according to claim 1,
The grid-
A cooling groove for inserting a grid holder for storing a plurality of the specimen grids to maintain the liquid holder in a state of being immersed in liquid nitrogen; And
A holder seating groove for withdrawing the grid holder from the cooling groove and seating the grid holder on one side of the grid receiving groove; And a workstation for cooperative measurement of an optical microscope and an electron microscope.
10. The method of claim 9,
Wherein the cooling groove and the holder seating groove are formed in a substantially rectangular shape,
And a protrusion of the concavo-convex structure is formed to take out the specimen grid from the grid holder, and a work station for the cooperative measurement of the optical microscope and the electron microscope.
The method according to claim 1,
And a lens groove into which a measurement lens capable of measuring a specimen grid accommodated therein is formed in a lower portion of the grid receiving groove.
The method according to claim 1,
The grid-
Further comprising a measurement lens holder for preventing the measurement lens from freezing or being frozen when the sample cooled by the liquid nitrogen is in close proximity to the measurement lens.
13. The method of claim 12,
The measurement lens holder includes:
A holder body into which the measurement lens is inserted;
A gas injection unit for injecting an inert gas into one side of the holder body; And
Wherein the measuring lens inserted in the holder body comprises a buffering agent for protecting against an external impact, and a workstation for cooperative measurement of an optical microscope and an electron microscope.
delete The method according to claim 1,
In the box portion,
The inner side is made of a thermally conductive material, the outer side is made of an acetal material,
And a heat insulating layer formed of a heat insulating material is formed between the inner side and the outer side in a multilayered structure. The work station for the cooperative measurement of an optical microscope and an electron microscope.
The method according to claim 1,
And a temperature sensor for measuring a temperature is formed inside the grid accommodation hole. The work station for coordinated measurement of an optical microscope and an electron microscope.
A workstation according to any one of claims 1 to 13 and 15 to 16 for performing a preprocessing process for cooling a specimen grid; And
A cooler in which liquid nitrogen is stored and an extension tube extending to one side of the cooler and connected to the work station during a preprocessing process and separated from the workstation after the pre- To be inserted into the measurement part of the specimen holder; And an electron microscope coupled to the optical microscope.
18. The method of claim 17,
Preferably,
The inner side is formed of a heat insulating material, the outer side is formed of an acetal material,
It is possible to adjust the thickness and length by providing a gear wheel inside,
And a detachable structure is formed in accordance with the size of the specimen mounting holder according to the size of the specimen mounting holder.
18. The method of claim 17,
In the specimen holder,
And a mounting head formed at the tip of the extension tube and having a grid mounting groove for mounting the specimen grid. The apparatus of claim 1,
18. The method of claim 17,
At the tip of the extension pipe,
And at least one step having a gradually smaller diameter is formed on the surface of the optical microscope.
18. The method of claim 17,
The workstation,
And a base plate for supporting the specimen mounting holder,
And a sliding guide having a stopper for sliding the specimen mounting holder in a direction away from the grid accommodating portion and limiting a depth at which the specimen mounting holder is inserted into the grid accommodating portion. The optical microscope and the electron microscope Linked imaging detection device.
18. The method of claim 17,
The workstation,
And a base plate for supporting the specimen mounting holder,
And a height adjusting device for supporting the lower surface of the base plate to adjust a vertical height of the work station.
23. The method of claim 22,
The height adjusting device comprising: a ground plate supporting the floor of the ground;
A folding member which is vertically driven by a plurality of links between the ground plate and the base plate; And
And a drive lever for vertically driving the folding member by inserting an actuating shaft screwed horizontally into the folding member. The linkage-type imaging detection device for linkage measurement of an optical microscope and an electron microscope.
A workstation according to any one of claims 1 to 13 and 15 to 16 for performing a preprocessing process for cooling a specimen grid;
A cooler in which liquid nitrogen is stored and an extension tube extending to one side of the cooler and connected to the work station during a preprocessing process and separated from the workstation after the pre- To be inserted into the measurement part of the specimen holder;
An optical microscope for measuring a signal for identifying an analyte on a specimen grid pretreated by the workstation; And
An electron microscope for analyzing the signal measured by the optical microscope; An interferometric imaging system for cooperative measurement of an optical microscope and an electron microscope.
25. The method of claim 24,
Characterized in that the cooler continuously supplies liquid nitrogen while the specimen mounting holder is detached from the work station and inserted into the electron microscope. ≪ Desc / Clms Page number 13 >
Placing a work station in a spaced apart position below the optical microscope in which a grid of specimens can be placed and having a grid receiving portion including a through hole formed therethrough at one side;
Inserting the specimen mounting holder through the through hole;
Placing a specimen grid in the grid housing and cooling the specimen grid;
Placing the cooled specimen grid in the specimen mounting holder;
Forming a signal on the cooled specimen grid using an optical microscope; And
And separating the specimen mounting holder from the work station after forming the signal, wherein the optical microscope and the electron microscope are combined.
27. The method of claim 26,
Further comprising the step of separating the specimen holder from the workstation and measuring the specimen holder with an electron microscope after the step of separating the specimen holder from the workstation.
27. The method of claim 26,
Wherein the step of placing the cooled specimen grid in the specimen mounting holder comprises the step of picking up the specimen grid cooled in the grid holder with a tweezers and transferring the specimen grid to the seating head.
27. The method of claim 26,
The sample holding holder is separated from the grid receiving part and then liquid nitrogen by the cooler provided in the sample mounting holder is supplied to the specimen grid until the measurement by the electron microscope is completed to maintain the cooling And an electron microscope.

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