KR101665057B1 - Spring-type cryo-plunger and Cooling method of sample grid using the same - Google Patents
Spring-type cryo-plunger and Cooling method of sample grid using the same Download PDFInfo
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- KR101665057B1 KR101665057B1 KR1020160060129A KR20160060129A KR101665057B1 KR 101665057 B1 KR101665057 B1 KR 101665057B1 KR 1020160060129 A KR1020160060129 A KR 1020160060129A KR 20160060129 A KR20160060129 A KR 20160060129A KR 101665057 B1 KR101665057 B1 KR 101665057B1
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- Prior art keywords
- sample
- cooling
- tweezers
- sample grid
- grid
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- 238000001816 cooling Methods 0.000 title claims abstract description 129
- 238000000034 method Methods 0.000 claims abstract description 54
- 238000003780 insertion Methods 0.000 claims abstract description 12
- 230000037431 insertion Effects 0.000 claims abstract description 12
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- 238000010168 coupling process Methods 0.000 claims description 27
- 238000005859 coupling reaction Methods 0.000 claims description 27
- 238000003860 storage Methods 0.000 claims description 20
- 239000000463 material Substances 0.000 claims description 18
- 238000007664 blowing Methods 0.000 claims description 2
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- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 10
- 229910052757 nitrogen Inorganic materials 0.000 description 5
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- OTMSDBZUPAUEDD-UHFFFAOYSA-N Ethane Chemical compound CC OTMSDBZUPAUEDD-UHFFFAOYSA-N 0.000 description 1
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- 239000002826 coolant Substances 0.000 description 1
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Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/02—Devices for withdrawing samples
- G01N1/10—Devices for withdrawing samples in the liquid or fluent state
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N25/00—Investigating or analyzing materials by the use of thermal means
- G01N25/14—Investigating or analyzing materials by the use of thermal means by using distillation, extraction, sublimation, condensation, freezing, or crystallisation
- G01N25/145—Accessories, e.g. cooling devices
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/416—Systems
- G01N27/447—Systems using electrophoresis
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/02—Devices for withdrawing samples
- G01N1/10—Devices for withdrawing samples in the liquid or fluent state
- G01N2001/1087—Categories of sampling
- G01N2001/1093—Composite sampling; Cumulative sampling
Abstract
The present invention relates to a spring-type ultra-low temperature sample plunger device and a method of cooling a sample grid using the same. The spring type ultra-low temperature sample plunger device according to an embodiment of the present invention includes a support portion and a support portion disposed on the support portion, A plunger portion, a rod portion, and a sample attaching portion, wherein the support portion includes a height adjusting portion for adjusting a height of the plunge portion, and the engaging portion of the sample attaching portion is a portion Wherein the cooling unit includes a groove, and a protrusion of the receiving unit is inserted into the cooling container, wherein the cooling unit includes a groove, a pin set insertion groove, a tweezers fixing unit for fixing the tweezers, and a tweezers fixing groove groove into which the tweezers fixing bar is inserted, Lt; / RTI > In addition, there may be provided a rotating table which is coupled to at least one surface of the receiving unit and rotates about a portion coupled to the receiving unit, a filter paper support member connected to the rotating table and capable of supporting a filter paper, And a blotting unit. Also, a humidity holding cover installed to enclose the main body from the outside and a humidity holding device connected to the humidity holding cover to supply steam into the humidity holding cover, a blotting time, and a sample grid, And a thermometer disposed on one side of the main body and sensing a temperature change during the sample grid cooling process. In this way, various tweezers and sample grids can be applied. Therefore, it is not necessary to replace the entire sample mounting part according to the types of the tweezers and the sample grid, so that the sample grid cooling can be easily performed and the sample grid blotting can be repeatedly performed at a precise position in a short time And the humidity of the sample grid cooling process can be kept constant, so that the success rate of the sample grid cooling process can be increased, and the temperature and time can be monitored and controlled in real time, so that the success rate of the sample grid cooling process .
Description
The present invention relates to a spring-type cryogenic sample plunger device and a cooling method of a sample grid using the same.
Generally, a transmission electron microscope (TEM) is a kind of electron microscope, which uses a light source and an electron lens instead of a light source lens, and has a very superior resolution compared to an optical microscope.
TEM has been used for research on the grain boundaries, interfaces, lattice defects, phase transitions, etc. of materials in materials science fields such as semiconductors and metals, In the field of biology, it is used for the study of living tissues such as microorganisms and cells, and for the analysis of the three-dimensional structure of protein complexes.
In particular, TEM has been effectively used for the study of biopolymers with small amounts of samples, and thus TEM is an indispensable device for the development of life sciences, new materials, and new functional devices.
Furthermore, the method of observing the sample using such a TEM is chemical fixation analysis, cryo analysis, heating analysis, and the like. First, the chemical fixation method is a method of observing a sample using a fixation solution in a fine cell structure And then fixed with a thin film.
However, this method has a disadvantage that it is difficult to observe the cell structure as it is originally, because the fixation fluid is slowly injected to the cell and the cell is deformed by the chemical fixative.
In addition, biological samples are generally difficult to visualize into a natural state due to damage by electron beams in the unstained state, and liquid samples can not be prevented from being dried in a vacuum microscope tube, Even if observation is possible, it is not possible to obtain an image suitable for the study by the movement of the sample.
On the other hand, water free biodegradation samples are very similar to natural samples because they are not influenced by chemical fixation or additional dyeing processes. Therefore, cryogenic analysis plays a very important role in life sciences research.
Here, in order to observe a biological sample by a cryogenic method, first, the sample is made into a liquid (water, buffer, etc.), and the sample is put on a predetermined pretreatment supporting membrane (grid) with a pipette, Sample grid is immersed in ethane solution cooled to liquid nitrogen temperature at high speed and cooled rapidly to prevent ice crystals from forming.
Subsequently, the rapidly cooled sample grid is transferred to a cryo transfer holder, and the holder is mounted on a TEM to obtain an image of the sample.
Here, in order to cool the sample grid to the liquid nitrogen temperature as described above, the sample grid must be immersed in liquid nitrogen using a separate apparatus. As the apparatus used at this time, for example, Vitrobot There is a cryo-sample plunger, such as Leica's EM GP.
More specifically, the conventional ultra-low temperature sample plunger devices as described above are generally configured such that a sample grid mounted on a holder is lowered using hydraulic pressure to immerse the sample grid in a container containing liquid nitrogen.
However, in the conventional ultra low temperature sample plunger devices as described above, in addition to the configuration for applying the hydraulic pressure, the configuration for controlling the hydraulic pressure must be separately provided, and the configuration is relatively complicated, The device itself was very expensive.
Therefore, in order to solve the problems of the prior art cryogenic sample plunger devices as described above, it is possible to provide a structure in which a function of immersing the sample grid in liquid nitrogen can be performed in a simpler configuration, for example, It is desirable to provide a cryogenic sample plunger device having a novel structure, but a device or a method that satisfies all of such requirements has not yet been provided.
The present invention can apply various tweezers and sample grids, can easily perform sample grid cooling, can perform sample grid blotting repeatedly at a precise position in a short time, It can increase the success rate of the sample grid cooling process by keeping the humidity constant during sample grid cooling process and it is possible to monitor and control the temperature and time in real time so as to increase the success rate of the sample grid cooling process. And an object of the present invention is to provide a cryogenic sample grid plunger device and a method of cooling a sample grid using the same.
A spring-type cryogenic sample plunger device according to an embodiment of the present invention includes: a body including a support portion and a support portion disposed on the support portion; A cooling container disposed on the support portion; A plunger portion connected to the support portion and disposed above the cooling vessel disposed on the support portion; A rod portion arranged to be movable up and down through the inside of the plunge portion; And a sample mounting part connected to the rod part and including a coupling part for coupling with the rod part and a tweezers for coupling to the coupling part, wherein the plug part includes a spring therein, The load section moves up and down and the support section includes a height adjusting section for adjusting the height of the plunge section, and the engaging section of the sample attaching section includes a tweezer insertion slot into which the tweezers are inserted, a tweezers fixed to the tweezers, Wherein the support portion includes a protrusion, and the cooling container includes a groove, and a protrusion of the support portion is coupled to the groove of the cooling container.
In addition, there may be provided a rotating table which is coupled to at least one surface of the receiving unit and rotates about a portion coupled to the receiving unit, a filter paper support member connected to the rotating table and capable of supporting a filter paper, The blotting portion may be disposed such that a filter paper disposed on the filter paper support is located at an end of the tweezers when the swivel is rotated to the upper portion of the receiving portion.
The apparatus may further include a moisture holding cover installed to surround the main body from outside and a humidity holding device connected to the humidity holding cover to supply humidity into the humidity holding cover.
The apparatus may further include a time measuring device disposed on one surface of the body to measure at least one of a blotting time and a time during which the sample grid is supported on the cooling material in the cooling container.
The apparatus may further include a thermometer disposed on one side of the main body and configured to detect a temperature change when the sample grid cooling process is performed.
A sample grid cooling method according to an embodiment of the present invention includes a main body including a support portion including a protrusion on an upper portion and a support portion disposed on the support portion, a cooling container including a groove on a lower surface, Preparing a spring-type ultra-low temperature sample plunger device including a rod portion arranged to be movable up and down through the inside of the plunge portion, and a sample mounting portion connected to the rod portion and coupled with the rod portion, ; Disposing the cooling container such that a projection of the receiving portion engages with a groove of the cooling container; Mounting a sample grid on the tweezers; Fixing the tweezers to the coupling part of the sample mounting part; Adjusting a height of the plunge portion by considering at least one of a length of the tweezers, a size of a sample grid, and a level of a cooling material in the cooling container by using a height adjusting portion of the supporting portion; And releasing the fixed state of the spring for fixing the spring installed on the plunge portion and lowering the rod portion.
The spring type ultra-low temperature sample plunger device may include a rotating table coupled to at least one surface of the receiving unit and rotating around a portion coupled to the receiving unit, a filter unit connected to the rotating table, The method comprising the steps of: mounting a filter paper on the filter paper support after adjusting a height of the plunge part; rotating the sweeper to an upper part of the receiving part Moving the filter paper on one side of the sample grid mounted on the tweezers, and blotting the sample grid using the filter paper.
The method may further include the step of maintaining a constant humidity in the sample grid cooling step using a humidity holding device.
The spring type ultra-low temperature sample plunger device according to the embodiment of the present invention and the cooling method of the sample grid using the same can be applied to various tweezers and sample grids. Therefore, it is not necessary to replace the entire sample mounting part according to the types of the tweezers and the sample grid, so that the sample grid cooling can be easily performed.
In addition, the sample grid blotting can be repeatedly performed at an accurate position in a short time, and can be easily performed.
In addition, it is possible to maintain the humidity constant during the sample grid cooling process, thereby increasing the success rate of the sample grid cooling process.
In addition, the temperature and time can be monitored and controlled in real time, which improves the success rate of the sample grid cooling process.
Figure 1 illustrates a spring-type cryogenic sample plunger device in accordance with an embodiment of the present invention.
Figure 2 shows a cooling vessel in a spring type cryogenic sample plunger device according to an embodiment of the present invention.
Figure 3 shows the cooling vessel.
Figures 4 and 5 illustrate a spring-type cryogenic sample plunger device in accordance with another embodiment of the present invention.
6 shows a sample mounting portion.
Fig. 7 (a) is a plan view of the sample mounting portion, and Fig. 7 (b) is a bottom view of the sample mounting portion.
Figs. 8 and 9 show the cooling container and its constitution.
10 shows a sample storage container.
FIG. 11 is a view illustrating a cooling type container in a spring-type cryogenic sample plunger device according to another embodiment of the present invention.
Hereinafter, preferred embodiments of the present invention will be described with reference to the accompanying drawings. However, the embodiments of the present invention can be modified into various other forms, and the scope of the present invention is not limited to the embodiments described below. Further, the embodiments of the present invention are provided to more fully explain the present invention to those skilled in the art. Accordingly, the shapes and sizes of the elements in the drawings may be exaggerated for clarity of description, and the elements denoted by the same reference numerals in the drawings are the same elements. In the drawings, like reference numerals are used throughout the drawings. In addition, "including" an element throughout the specification does not exclude other elements unless specifically stated to the contrary.
Spring type cryogenic sample plunger Device
Hereinafter, a spring type ultra-low temperature
Figure 1 illustrates a spring-type cryogenic
1 to 3, a spring-type cryogenic
The body may include a receiving portion (18) and a supporting portion (11). The
The supporting
Alternatively, the supporting
The
The spring type ultra low temperature
Referring to Figs. 2 and 3, the receiving
The
The apparatus may further include an
The apparatus may further include a
Fig. 6 shows a
6 and 7, the
The engaging
The
The pincette fixing groove may be a
Figs. 8 and 9 show the cooling
8 and 9, the
The cooling
In the spring type cryogenic
After the cooling of the sample grid in the cooling
And a
The
In addition, by forming the vibration ditch at a certain depth in the bottom surface portion where the cooling
Through the configuration of the spring type ultra-low temperature
After cooling of the sample grid in the cooling
In addition, by implementing the spring type ultra-low temperature
In addition, according to the present invention, the spring type ultra-low temperature
Figures 4 and 5 illustrate a spring type cryogenic
4 and 5, a spring type ultra-low temperature
Blotting is a process that is carried out before the sample grid is immersed in the cooling material. In this process, the liquid on the sample grid is partially absorbed through the filter paper in order to leave only a certain amount of the liquid contained in the sample grid. to be. Therefore, it is important to stably approach the filter paper to at least one side of the sample grid mounted on the
Since the spring type ultra low temperature
The
FIG. 11 shows a
11, the spring type ultra-low temperature
The moisture-retaining
11, a spring type ultra-low temperature
The blotting time of the sample grid and the time the sample grid is immersed in the cooling material should be precisely controlled. If the blotting is short or long, the sample grid may not be broken or cooled due to a large amount of liquid remaining on the sample grid or remaining on the sample grid. In addition, if the sample is not immersed in the cooling material for a certain period of time, the sample grid may not be cooled.
11, a spring type ultra-low temperature
Sample Grid Cooling Method
Hereinafter, a sample grid cooling method according to an embodiment of the present invention will be described.
A method of cooling a sample grid according to an embodiment of the present invention includes a main body including a support portion 18 including a projection 18a on an upper portion and a support portion 11 disposed on the support portion 18, A plunger portion 12 connected to the support portion 11, a rod portion 13 arranged to be movable up and down through the inside of the plunger portion 12, Preparing a spring-type cryogenic sample plunger device (10) including a sample mounting portion (14) connected to the rod portion (13) and including a coupling portion (31) that engages with the rod portion (13); Disposing the cooling vessel (15) so that the projection (18a) of the receiving portion (18) engages with the groove (15a) of the cooling vessel (15); Mounting a sample grid on the tweezers (32); Fixing the tweezers (32) to the coupling part (31) of the sample mounting part (14); Considering the length of the tweezers 32, the size of the sample grid, and the level of the cooling material in the cooling vessel 15, using the height adjusting unit 11c of the supporting unit 11, (12); And releasing the fixed state of the spring for fixing the spring provided on the plunge part (12) and lowering the rod part (13).
In the step of preparing the spring type cryogenic
In the step of disposing the cooling
In the step of mounting the sample grid on the
The
The height of the plunge part (32) may be adjusted by considering the length of the tweezers (32), the size of the sample, and the level of the cooling material in the cooling vessel (15) 12, the height of the
The sample grid can be immersed in the cooling material in the cooling
In the sample grid cooling method according to the embodiment of the present invention, the spring type ultra-low temperature
The blotting step may include a back-blotting step in which the
The sample grid cooling method according to the embodiment of the present invention may further include the step of keeping the humidity constant during the sample grid cooling process using the
The present invention is not limited to the above-described embodiment and the accompanying drawings, but is intended to be limited by the appended claims. It will be apparent to those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. something to do.
10: Spring type cryogenic sample plunger device
11: Support
11a:
11b: Inner support
11c:
12: Plunge part
13:
14: sample mounting part
15: cooling vessel
15a: groove of the cooling vessel
16:
17: chamber
18:
18a: projection of the pedestal portion
19: Blowing part
19a:
19b: Filter paddle support
21: Humidity maintenance cover
22: Humidity holding device
23: Time measuring device
24: Thermometer
31:
32: Tweezers
41: Rod insertion groove
42:
43: Tweezers insertion groove
44: Bolt
51: inner container
52: outer container
53: sample holder
54: Cradle
61: Heat dissipation means
71: Sample storage container
72: Sample storage groove
73: cover
Claims (22)
A cooling container disposed on the support portion;
A plunger portion connected to the support portion and disposed above the cooling vessel disposed on the support portion;
A rod portion arranged to be movable up and down through the inside of the plunge portion; And
A sample mounting part connected to the rod part and including a coupling part for coupling with the rod part and a tweezers for coupling with the coupling part; And
And a blotting unit coupled to at least one side of the receiving unit and supporting a filter paper,
Wherein the blotting unit includes a rotating unit that rotates about a portion coupled to the receiving unit, a filter paper support that is connected to the rotating unit and is capable of supporting a filter paper,
Wherein the blowing portion is disposed such that the filter paper disposed on the filter paper support is located at an end of the tweezers when the swivel is rotated to the upper portion of the receiving portion.
And a moisture holding device connected to the humidity holding cover to supply steam into the humidity holding cover. The apparatus of claim 1, further comprising:
Further comprising a time measuring device disposed on one side of the main body for measuring at least one of a blotting time and a time during which the sample grid is supported on the cooling material in the cooling vessel, .
Further comprising a thermometer disposed on one side of the main body to detect a temperature change when performing a sample grid cooling process.
Wherein the cooling container includes an outer container, an inner container disposed inside the outer container, and a cradle for receiving the outer container at an upper portion thereof.
Wherein the cradle includes a sample mounting part for mounting the sample grid.
And a sample storage container disposed on the sample holder.
Further comprising a radiating means provided in a bridge shape in contact with an upper portion of the outer container and an outer surface of the cradle.
Further comprising: a housing disposed to surround the inner container, the holder, and the heat dissipating means; and a cover disposed at an upper portion of the housing.
Further comprising an operation control part serving as a switch for fixing the spring provided on the plunge part and releasing the fixed state of the spring for lowering the rod part.
Wherein the sample mounting portion includes a coupling portion for coupling to the rod portion and a tweezers inserted and fixed at a lower end of the coupling portion to fix the sample grid.
Further comprising a chamber extending downwardly from a lower end of the plunger to prevent humidity control of the sample grid during cooling and splashing of the cooling solution to the user.
Wherein the coupling portion includes a rod insertion groove formed on an upper surface for coupling to the rod portion and a screw hole formed in the center of the rod insertion groove.
Wherein the fixing groove of the tweezers is a screw hole formed in a side surface of the coupling portion, and the fixing base of the tweezers is a bolt inserted into the screw hole.
Wherein the sample storage container comprises a plurality of sample storage grooves formed on the upper surface of the cylindrical body and a cover configured to be capable of opening and closing the sample storage grooves by a rotary type so that the cooled sample grids are transferred to the respective sample storage grooves And then the sample grid is moved and conveyed after closing the cover.
The dust container may further include a dustproof groove formed at a bottom portion of the bottom of the cooling container. The cooling container may be installed in the dustproof groove to prevent vibration due to the falling of the rod Features a spring-type cryogenic sample plunger device.
Wherein after the cooling of the sample grid is completed in the cooling vessel, the cooled sample grids are placed in a sample holder portion provided in the cooling vessel, thereby cooling the plurality of sample grids while maintaining the temperature of the cooled sample grids Of the spring type cryogenic sample plunger device.
Disposing the cooling container such that a projection of the receiving portion engages with a groove of the cooling container;
Mounting a sample grid on the tweezers;
Fixing the tweezers to the coupling part of the sample mounting part;
Adjusting a height of the plunge portion by considering at least one of a length of the tweezers, a size of a sample grid, and a level of a cooling material in the cooling container by using a height adjusting portion of the supporting portion;
The method comprising the steps of: attaching a filter paper to the filter paper support base; rotating the rotation base to an upper portion of the receiving unit to move the filter paper onto one surface of a sample grid mounted on the tweezers; Blotting the surface of the substrate; And
And fixing the spring installed on the plunge portion and releasing the fixed state of the spring for lowering the rod portion.
Wherein the blotting step blotting is performed by contacting a rear surface of a portion of the sample grid where the sample is disposed.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020160060129A KR101665057B1 (en) | 2016-05-17 | 2016-05-17 | Spring-type cryo-plunger and Cooling method of sample grid using the same |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020160060129A KR101665057B1 (en) | 2016-05-17 | 2016-05-17 | Spring-type cryo-plunger and Cooling method of sample grid using the same |
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| KR101665057B1 true KR101665057B1 (en) | 2016-10-24 |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2020013499A1 (en) * | 2018-07-12 | 2020-01-16 | 서울대학교산학협력단 | Apparatus for manufacturing electron microscope grid sample |
| KR20200007641A (en) * | 2018-07-12 | 2020-01-22 | 서울대학교산학협력단 | Manufacturing apparatus of electron microscopy grids sample |
| WO2023219401A1 (en) * | 2022-05-10 | 2023-11-16 | 주식회사 파이로솔루션 | Filtering device for microplastic analysis using py-gc/ms |
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| KR101388509B1 (en) | 2013-12-06 | 2014-04-28 | 한국기초과학지원연구원 | Spring-type cryo-plunger |
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- 2016-05-17 KR KR1020160060129A patent/KR101665057B1/en active IP Right Grant
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| KR200442697Y1 (en) | 2007-04-06 | 2008-12-05 | 서성교 | A cryo- rack |
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| KR101388509B1 (en) | 2013-12-06 | 2014-04-28 | 한국기초과학지원연구원 | Spring-type cryo-plunger |
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