JP3356636B2 - Quick freezing device - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a method for preparing a sample for observing a biological sample using a transmission electron microscope, for example, by briefly pressing a biological sample against a metal block cooled to liquid helium temperature. The present invention relates to a quick freezing apparatus for rapidly freezing and fixing.

[0002]

2. Description of the Related Art The basic structure of a quick freezing apparatus of this type is introduced in, for example, "Experimental Medicine", Vol. 8, No. 5, pp. 152 to 158, and "Electron Microscope", Vol. 19, No. 2, 135. From page 138 to page 138, a configuration in which contamination between samples during the freezing of samples is prevented is introduced. Fig. 10 shows the quick-freezing device introduced in the above-mentioned "Experimental Medicine"
This will be described with reference to FIG. In FIG. 10, 1 is a liquid nitrogen container, 2 is a liquid helium container, 3 is liquid nitrogen, 4 is liquid helium, 5 is a base cooling block, 6 is a metal block, 7 is a liquid helium transfer line, 8 is a decompression duct, 9 Denotes a shutter, 10 denotes a sample insertion rod, 11 denotes a sample, 12 denotes a sample holder, 13 denotes an insertion guide cylinder, and 14 denotes a heater. Reference numeral 100 denotes an upper surface base of the liquid nitrogen container and the liquid helium container. The liquid helium transfer line 7, the pressure reducing ducts 8 and 9 use the upper surface base 100 to form the shutter 9, the insertion guide cylinder 13, the heater 14, and the like.
Etc. are supported. The base cooling block 5 is held at the bottom of the liquid helium container 2, and the metal block 6 is supported on the base cooling block 5 so as to be detachable. Reference numeral 200 denotes a housing having a heat insulating layer on the inner surface for housing the liquid nitrogen container and the liquid helium container.

A liquid helium container 2 is provided inside a liquid nitrogen container 1, and each container is filled with liquid nitrogen 3 and liquid helium 4, respectively. A detachable metal block 6 is mounted on the base cooling block 5 immersed in the liquid helium in the liquid helium container 2 and is in thermal contact to maintain the liquid helium temperature. The liquid helium is supplied from the liquid helium transfer line 7 and recovers the liquid helium evaporating gas through the decompression duct 8 or performs vacuum evacuation when filling the liquid helium. The shutter 9 prevents the outside air from entering the liquid helium container 2.
Is electrically driven by the insertion of the, and is opened.

[0004] In a freezing operation of a biological sample, a sample 11 is attached to a sample holder 12 through a filter paper or a buffer material, and a sample insertion rod 10 is attached.
Attach to the tip of. The insertion of the sample insertion rod 10 is manually performed by using the insertion guide tube 13 as a guide. A heater 14 is attached to the insertion guide tube 13, and the inside of the insertion guide tube 13 is controlled to a temperature slightly higher than room temperature.
After being brought into contact with the metal block 6 for an arbitrary time, the frozen sample is promptly taken out and immersed in liquid nitrogen or a chemical fixing solution to preserve the frozen sample.

[0005] In the above-mentioned "electron microscope", the sample contact surface of the metal block 6 is further shifted slightly, so that the metal block 6 is replaced every time the sample is frozen while the new contact surface remains. It introduces an example devised so that it is not necessary.

[0006]

In the above-mentioned conventional freezing operation of a biological sample, an insertion crimping operation is performed manually.
The descending speed and crimping time of the sample insertion rod 10 vary depending on the experience of the operator or the dexterity of the operator, and it is difficult to obtain a frozen sample with good reproducibility. Further, the operation of replacing the metal block 6 after the crimping surface of the metal block 6 for crimping the biological sample is contaminated by the body fluid of the biological tissue by the crimping has not been sufficiently considered. For this reason, evaporation of liquid helium accompanying the exchange operation has been accelerated.

A good frozen biological sample in which ice crystals generated in cells and tissues are kept as small as possible and structural destruction due to the growth of ice crystals is minimized is, for example, 1000.
A freezing speed of 0 ° C./sec or more is required, and a good freezing surface of 15 to 20 μm or more can be obtained from the freezing surface, but this is stably realized by the operation of the operator. It was difficult. Furthermore, it is difficult to irradiate the sample with light immediately before the freezing operation of the sample to cause a dynamic change in the place where various functions in the biological tissue cells exist due to the optical information signal, and to freeze and fix it. there were.

[0008]

SUMMARY OF THE INVENTION According to the present invention, a sample freezing operation is sequenced and automated, and a metal block cooled with liquid helium can be exchanged through an exchange port having a limited opening area. Accordingly, it is an object of the present invention to realize a quick freezing apparatus with good reproducibility of the freezing operation regardless of the experience of the operator. It is another object of the present invention to realize a quick freezing apparatus in which loss of liquid helium is small even when replacing a metal block. Further, by incorporating a process of irradiating light having different wavelengths and light amounts in a specific region immediately before the biological sample is rapidly frozen in the sequence, the location where various functions in the biological tissue cell by the optical information signal are present is provided. It is an object of the present invention to provide a quick freezing device which is capable of freezing and fixing a dynamic change at a time.

[0009]

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS An embodiment of the present invention will be described below with reference to the drawings.

FIG. 1 is a diagram showing a schematic configuration of an embodiment of a quick freezing apparatus according to the present invention. 15 is a liquid nitrogen container, 16 is a liquid nitrogen, 17 is a liquid helium container, 18 is liquid helium, 19 is a liquid helium transfer tube, 20
Is a liquid nitrogen filling port, 21 is a base cooling block,
22 is a metal block, 23 is a base cooling block rotary gear, 50 is a rotary drive mechanism, 24 is a rotary drive rod, 25 is a rotary drive gear, 26 is a shutter, 27 is a heat retaining cylinder, 28 is a heater, 29 is a sample insertion rod, Reference numeral 30 denotes a sample, 31 denotes a sample holder, 32 denotes a moving mechanism, 33 denotes a sample collecting container, 47 denotes a rotating mechanism of the sample collecting container, 34 denotes a push rod, 36 denotes a moving mechanism support, and 35 denotes a control device. Numeral 39 denotes a holding mechanism for the moving mechanism supporting portion 36, which can hold the moving mechanism supporting portion 36 in a state where the moving mechanism supporting portion 36 is inclined by an arbitrary angle from the vertical state shown in the drawing. 62 is an upper surface base, 200 is a housing | casing which accommodates a liquid nitrogen container and a liquid helium container.

A liquid helium container 17 is provided inside a liquid nitrogen container 15, and each container is filled with liquid nitrogen 16 and liquid helium 18. The liquid helium 18 is supplied from the liquid helium transfer tube 19 to the liquid nitrogen 1.
6 are filled from liquid nitrogen filling ports 20, respectively. A detachable metal block 22 is mounted on the base cooling block 21 immersed in the liquid helium 18 in the liquid helium container 17, and the liquid helium temperature is maintained by thermal contact. Further, a metal block rotating gear 23 is attached to the base cooling block 21 to rotate the rotation driving mechanism 2.
4 transmits a rotational force via a rotary drive gear 25. This means that every time the biological sample is frozen, the metal block 2
This is because the crimping surface of No. 2 is contaminated with the body fluid of the biological tissue by the crimping, while the crimping surface is rotated to a new crimping surface. Shutter 26 provided directly above metal block 22
A heater 28 is provided on the inner surface of the attached heat retaining cylinder 27, and the inside of the heat retaining cylinder 27 is controlled to a temperature slightly higher than room temperature. The shutter 26 prevents outside air from entering the liquid helium container 17 and keeps the sample at room temperature until just before the sample is frozen.

The sample holder 31 is attached to the tip of the sample insertion rod 29.
The sample insertion rod 29 is lowered into the liquid helium container 17 by the moving mechanism 32, and the sample 30 is pressed against the metal block 22. After a short time, the sample insertion rod 29 is Obtaining the frozen sample by pulling it out of the container 17 is essentially the same as the device described in the above-mentioned document, but in the present invention, these operations are sequenced by the controller 35. Although not shown in FIG. 1 in order to avoid complicating the drawing, it is devised to reduce the loss of liquid helium when the metal block 22 is replaced.

FIG. 2 is a view for explaining the operation state of the freezing mechanism of the embodiment of the quick freezing apparatus according to the present invention, and (a).
3B shows a state in which the sample insertion rod 29 descends to press the sample against the metal block 22 and freezes, and FIG. 4B shows a state in which the sample insertion rod 29 rises and collects the sample.

The holding mechanism 39 allows the moving mechanism support 3
6 is held at an arbitrary angle, a sample holder 31 is attached to the tip of the sample insertion rod 29, and the sample holder 30 is attached thereto.
Is set. At this time, the sample 30 is attached to the sample holder 31 via a filter paper or a cushioning material. A thermally insulated chuck is provided at the tip of the sample insertion rod 29, and the sample holder 31 is attached to the chuck. However, the illustration of the chuck is omitted for simplification of the drawing. Thereafter, the moving mechanism support section 36 is returned to the vertical state, and a start button (not shown) of the control device 35 is pressed. As a result, as shown in FIG. 2A, the sample insertion rod 29 descends by pneumatic drive to press the sample 30 against the metal block 22 as described later, and the sample is frozen. During the freezing operation of the sample, prior to the lowering of the sample insertion rod 29, the sample collection container 33 is moved to a position where the sample collection container 33 does not collide (a retreat position) by the rotation mechanism 47 of the sample collection container.

As is apparent from FIG.
The shutter 26 is mechanically opened by the insertion of the sample, but the sample mounted on the tip of the sample insertion rod 29 passes through the heat retaining cylinder 27 heated by the heater 28 until immediately before being pressed against the metal block 22. The metal block 22
The temperature change of the sample in contact with is extremely large. As described above, by maintaining the temperature immediately above the metal block 22 at a temperature slightly higher than room temperature so that a large temperature gradient can be obtained during the freezing operation of the sample, a good freezing state can be achieved without causing the sample to crystallize. The resulting freezing rate, eg, 10,000 ° C./sec or more, can be easily achieved. Furthermore, by opening the shutter 26 immediately before pressing, the leakage of warm air into the liquid helium container can be reduced, and the loss of liquid helium can be reduced.

When a predetermined time has elapsed, FIG.
As shown in the figure, the sample insertion rod 29 is automatically returned to the raised position by pneumatic driving, contrary to the freezing operation.
In response to this return, the sample collection container 33 is moved to a position immediately below the sample holder 31 as shown in the figure, and then the sample 30 is pushed out together with the sample holder 31 into the sample collection container 33 by the push rod 34, and The frozen sample is stored by immersion in nitrogen or a chemical fixative. here,
When the sequence from the position control of the sample collection container 33 to the collection by pushing the sample holder 31 and the sample 30 into the sample collection container 33 by the push rod 34 is sequenced, the sample 3
Good storage of the frozen sample is performed immediately after the 0 is crimp-frozen.

It is to be understood that the sample insertion rod 29
The shutter 26 is mechanically interlocked and closed in response to the return to the raised position. Then, as described later, the metal block 22 is rotated to prepare for the next sample freezing.

FIG. 3 is a view for explaining the operation state of the rotating mechanism of the metal block of the embodiment of the quick freezing apparatus of the present invention.
(A) is a plan view of the metal block rotation mechanism, and (b) is a cross-sectional view of the metal block rotation mechanism.

The metal block 22 is driven by a rotary drive gear 25 via a rotary drive rod 24 by a rotary drive mechanism 50. Numeral 30 indicates a sample pressed on the metal block 22, and another circle shown by a broken line indicates a region used for another sample. In this example, one metal block 22 can be used for eight samples.

FIGS. 4 and 5 are a sectional view and a plan view, respectively, showing the relationship between a port for attaching and detaching the cooling metal block 22 and a port for inserting a frozen sample in the embodiment of the quick freezing apparatus.

As shown in FIGS. 4 and 5, although omitted in FIG. 1 for simplification of the drawing, a port switching mechanism 51 is provided immediately below the insertion rod 29 in the present invention. The port switching mechanism 51 of the present embodiment includes a metal block exchange port 52 and a sample insertion port 53 and is integrally formed, and is devised so that switching can be performed by simple rotation. An outside air blocking cover 54 is always fitted to the metal block exchange port 52, and the opening of the sample insertion port 53 is always closed by the shutter 26 as described with reference to FIGS. Therefore, in the preparation stage for freezing the sample, the leakage of warm air into the liquid helium container can be prevented, and the loss of liquid helium can be reduced. During the sample freezing operation, the sample insertion port 53 is located immediately below the insertion rod 29.
The sample is frozen as described with reference to FIG. FIG. 4 simply shows the state at this time. As described above, the port switching mechanism 5 of the present embodiment
In FIG. 5, a metal block exchange port 52 and a sample insertion port 53 are integrally formed and devised so that they can be switched by simple rotation. Therefore, as shown in FIG. 5 (the lid 54 is omitted here) as shown in FIG. With the insertion rod 29 pulled out, the port switching mechanism 51 is moved to the
When rotated by 80 degrees, the metal block exchange port 52 comes directly below the insertion rod 29 as indicated by a broken line 52 '. In other words, the metal block exchange port 52 comes directly above the metal block 22. In this state, if the metal block 22 is exchanged with an exchanging tool as described later, it is possible to reduce the inflow of outside air into the helium container from the port at the time of exchanging the metal block. In FIG. 5, reference numeral 50 indicated by a broken line denotes a rotary drive mechanism;
Indicates a rotation mechanism of the sample collection container, and an arrow indicates a state of rotation of the sample collection container 33.

That is, in the present embodiment, the entry of outside air through the respective ports can be suppressed to a minimum during the sample freezing operation and during the replacement of the metal block. The metal block exchange port 52 and the sample insertion port 53 do not need to be integrally configured as shown in the figure. For example, the sample insertion port 53 is arranged on the upper surface of the metal block exchange port 52. , Usually metal block exchange port 52
May be closed by the sample insertion port 53, and the sample insertion port 53 may be shifted so that the opening of the metal block exchange port 52 appears when replacing the metal block.

FIG. 6 is a diagram for explaining an example of drive sequence control of the embodiment of the quick freezing apparatus according to the present invention. FIG. 6 is slightly different from FIG. 1 in order to make it easier to see the entire movement comprehensively, but the relative configuration of each component is the same.

Numeral 36 denotes a moving mechanism supporting portion. In this embodiment, a cylinder portion having a piston-cylinder structure is used as the moving mechanism supporting portion as it is.
2 is mounted so that it can move up and down. An arrow 37 indicates the flow of the compressed air, which is supplied from an air source (not shown). 38a to 38f are solenoid valves,
Each is opened and closed by the control device 35 according to a predetermined sequence. Here, the solenoid valves 38a and 38d are not simple open / close, but can change the output port of the compressed air in response to the operation signal. Reference numerals 40 to 43 denote position sensors which detect the position of the moving mechanism 32, respectively. Reference numeral 40 denotes an ascending position, 41 denotes a descending position, and 42 and 43 denote intermediate position detecting sensors. The detection signal of each position sensor is sent to the control device 35. As shown in the figure, the state where the moving mechanism 32 is close to the raised position is the initial state of the apparatus,
In this state, the sample holder 31 is attached to the tip of the sample insertion rod 29, and the sample 30 is set on the holder. When the start instruction is given to the controller 35 after the sample 30 is set,
The electromagnetic valve 38a is opened and the compressed air is moved by the moving mechanism support 3
6. The moving mechanism 32 is supplied to the upper part of the piston 6 and pushes the moving mechanism 32 downward. This naturally pushes down the sample insertion rod 29 at the same speed. When the piston reaches the lowered position,
A detection signal is sent from the sensor 42 to the control device 35, the electromagnetic valve 38a is closed, and the moving mechanism 32 is held at that position.
That is, the sample 30 is kept pressed against the metal block 22. When a predetermined time set in the control device 35 elapses, a signal is sent from the control device 35 to the solenoid valve 38a, and compressed air is supplied to the lower portion of the piston of the moving mechanism support portion 36 to move the moving mechanism 32 upward. Push up. When the piston reaches the raised position, a detection signal is sent from the sensor 40 to the control device 35, the electromagnetic valve 38a is closed, and the moving mechanism 32 is held at that position and returned to the initial state. The speed at which the sample insertion rod 29 is depressed can be determined in advance by using the intermediate position sensors 42 and 43 to determine the relationship between the solenoid valve opening and the speed. If you set it, you can insert at the desired speed. When the frozen sample is manually collected, the sample is collected together with the sample holder 31 by pushing the push rod 34 as described above.

Next, not only the sequence of the freezing operation, but also the sequence including the sample recovery after freezing and the device for reducing the intrusion of outside air into the helium container 17 during the freezing operation will be described.

First, before lowering the sample insertion rod 29, it is necessary to place the sample collection container 33 in the position shown by the broken line in FIG. For example, when the power of the apparatus is turned on, the sample insertion rod 29 (moving mechanism 32) is brought to the raised position. At this time, the electromagnetic valve 38d is opened and the rotary cylinder 47 is driven to drive the sample collection container. Control is performed so that the position of 33 is a position indicated by a broken line in FIG. The rotary cylinder 47 is provided with position sensors 48 and 49 for detecting a position immediately below and a retracted position, respectively. After that, when the start instruction is given, the sample insertion rod 29 is lowered, and after the sample is frozen, the sample insertion rod 29 is pulled up, and when the sample insertion rod 29 is at the raised position, the solenoid valve 38d is opened. To drive the rotary cylinder 47. At this time, the position of the sample collection container 33 is the position shown in FIG.
As shown in (b), the position immediately below the sample insertion rod 29 (4
(8 outputs). Thereafter, after a predetermined time has elapsed, a command to open the electromagnetic valve 38b is given from the control device 35. Therefore, compressed air is added to the upper surface of the piston portion of the push rod 34 to push down the push rod 34, and the sample holder 31 and the sample 30 are pushed out. At this time, since the sample collection container 33 is located immediately below the sample sample insertion rod 29, the frozen sample is pushed out to the sample collection container 33 and collected. After a relatively short time, the solenoid valve 38b is closed and the compressed air on the upper surface of the piston portion of the push rod 34 is exhausted, so that the pushed down push rod 34 is repelled by a contracting spring (not shown). Return to initial position. At about the same time, the control device 35 opens the solenoid valve 38d for a short time and drives the rotary cylinder 50 to drive the metal block 22 because the sample insertion rod 29 is in the raised position.
Is rotated by a predetermined rotation angle so that the next clean surface indicated by the broken line in FIG. In this state, the device is again the same as in the initial state.

In this way, after the sample is set, the operator simply instructs start, the entire process up to the collection is sequenced, and the frozen sample can be stored well immediately after the sample is crimp-frozen. Done.

Further, the sample insertion rod 29 descends and the heat retaining cylinder 2
When the shutter 26 is opened during the festival passing through the inside of the inside 7, it is effective to prevent the outside air in the heat retaining tube 27 from being discharged into the liquid helium container by the sample insertion rod 29. Therefore, in the present embodiment, when a predetermined time has elapsed after the sample insertion rod 29 has begun to descend, or when the position sensor 42 has output, the electromagnetic valve 38e is opened to drive the ejector 44b to drive the ejector 44b into the heat retaining cylinder. The outside air is sucked and exhausted together with the liquid helium vaporized gas 45. In addition, the heat insulation cylinder 27
Is provided with a temperature sensor 61, which sends an output signal to the control device 35 and controls the heater 28 using the output signal.

Further, a thermally insulated chuck 44 is provided at the tip of the sample insertion rod 29, and the sample holder 31 is attached. Chuck 4 for holding sample holder 31
Although not shown, a notch 4 is provided around the periphery to facilitate the attachment / detachment of the sample holder 31 and to assist the gas flow around the sample holder 31 and the sample 30. That is, when the insertion rod 29 is lowered and the sample 30 is pressed against the metal block 22, the helium evaporating gas 45 in the liquid helium container is forcibly flowed around the sample holder 31 and the sample 30, so that the sample Can improve the freezing effect. For this reason, similarly to the solenoid valve 38e, when a predetermined time elapses after the sample insertion rod 29 starts to descend,
Alternatively, when the position sensor 42 outputs, the electromagnetic valve 38
is opened, the ejector 44a is driven, and the sample insertion rod 2 is opened.
The outside air of a waste tube (not shown) communicating with the inside 9 is sucked and exhausted. When the sample 30 is pressed against the metal block 22, the helium vaporized gas 45 in the liquid helium container is immediately prepared to be forced to flow around the sample holder 31 and the sample 30. 2
The liquid helium evaporating gas 45 continues to flow around the sample 30 through the inside of the sample insertion rod 29 by continuously driving the ejector 44 a while being pressed against the sample 2.

FIG. 7 is a time chart showing the control of the embodiment of the quick freezing apparatus according to the present invention. As is clear from the drawings, in this embodiment, all operations are performed according to a predetermined sequence only by the operator instructing start after mounting the sample. In this embodiment, the moving speed of the moving mechanism is 1.5 m / sec at the maximum.

FIG. 8 is a schematic configuration diagram of an embodiment of a quick freezing apparatus which enables a sample to be irradiated with light immediately before freezing of the sample in the embodiment of the quick freezing apparatus. As compared with the embodiment of FIG. 1, a light irradiation device 55 capable of irradiating a sample mounted on the tip of the sample insertion rod 29 with light of a desired wavelength is provided. In this embodiment, the start instruction is given by the operator, and at the same time, the control device 35 gives the instruction of light signal irradiation. When the amount of light is insufficient, the descent of the sample insertion rod 29 may be started slightly after the start instruction.

The living body recognizes light of a specific range of wavelength and light amount as environmental information by using optical sensors having several different functions, and adjusts various functions in the living body. In order to elucidate the mechanism of an optical switch that controls gene expression in response to this optical information and to elucidate the information transmission in cells, light of a specific wavelength (red light: wavelength 5
50-690 nm, near infrared light: wavelength range 700-800 n
m, blue light: a wavelength range of 400 to 480 nm) is provided with a light irradiation device 51 for individually irradiating the light amount and time or by providing a plurality of lights in different wavelength ranges, and a “trigger type optical switch” depending on the light irradiation conditions. Fight chrome A or "on
The "off-type optical switch" is used to freeze-fix immediately after changing the specificity of biological cells / tissues and the dynamics of the signal transmission system using different optical switches such as phytochrome B and cryptochrome.

FIG. 9 shows a liquid nitrogen pre-cooling exchange mechanism of the metal block 22 for freezing the sample on the surface of the embodiment of the rapid freezing apparatus.

As shown in FIG. 3, the metal block 22 needs to be replaced after freezing eight samples even if the metal block 22 is used while shifting the clean surface. At this time, when the replacement metal block 22 is taken directly from the outside air into the helium container 17 and loaded on the base block 21 cooled with liquid helium, the helium in the helium container 17 is strongly evaporated. At the same time, there is a possibility that water vapor from outside air freezes on the upper surface of the metal block 22. FIG. 9 shows a configuration example for performing replacement after pre-cooling the metal block 22 to be newly replaced. The metal block 22 for pre-cooling is mounted on the base cooling block 60 cooled by the liquid nitrogen 59 in the liquid nitrogen container 58 and cooled. When replacing the metal block, the metal block 22 which has been prestressed on the base cooling block 60 is gripped by the metal block replacement rod 57 and is held in the metal block replacement port 5 described with reference to FIG.
2 and carried into the helium container 17 and loaded on the base block 21 cooled with liquid helium. At this time, dry helium gas 56 is applied to the metal block replacement rod 57 so that frost does not adhere to the upper surface of the metal block 22.
It is effective to wrap the metal block 22 with the dry helium gas 56 to prevent frost from adhering by introducing the gas through the tube and flowing it out from the tip of the replacement rod 57. The liquid nitrogen container 58 for cooling the base cooling block 60 in this embodiment may be provided independently, but as will be easily understood with reference to FIG.
Since there is a liquid nitrogen container 15 in 00, it is supposed that the upper surface of the container is opened with respect to the upper surface base 62 so as to be used as shown in FIG. There is an advantage that a simple device can be obtained.

In this embodiment, the operation of the moving mechanism is performed by air pressure. However, it is obvious that the moving mechanism may be electrically driven. For example, a linear motor or a plunger type driving device may be used. Available.

According to the present invention, the fine structure of cells and tissues can be fixed and preserved as it is, and the dynamic moment of life activity can be captured. Further, since no chemical treatment is required as a pretreatment for freezing, there is little outflow and deformation of intracellular substances, and artificially generated products such as structural deformation or destruction due to chemical substances can be avoided. In addition, good (glass-like freezing) essential for preservation of microstructure can be obtained.

[0037]

As is apparent from the above description, according to the present invention, free and reproducible freezing can be obtained without requiring any skill in the operation, and the efficiency of the freezing operation can be improved.

[Brief description of the drawings]

FIG. 1 is a diagram showing a schematic configuration of an embodiment of a quick freezing apparatus of the present invention.

FIG. 2 is a diagram illustrating an operation state of an embodiment of the quick freezing apparatus of the present invention.

FIG. 3 is a diagram illustrating an operation state of a rotation mechanism of the quick freezing apparatus according to the embodiment of the present invention.

FIG. 4 is a diagram illustrating a cross section of a mechanism for switching ports in the embodiment of the quick freezing apparatus of the present invention.

FIG. 5 is a diagram illustrating a plane of a mechanism for switching ports in the embodiment of the quick freezing apparatus of the present invention.

FIG. 6 is a view for explaining drive control of the embodiment of the quick freezing apparatus of the present invention.

FIG. 7 is a time chart for explaining control of the embodiment of the quick freezing apparatus of the present invention.

FIG. 8 is a diagram showing a schematic configuration of a quick freezing apparatus provided with a light irradiation device according to an embodiment of the quick freezing apparatus of the present invention.

FIG. 9 is a diagram showing a schematic configuration of a metal block liquid nitrogen precooling mechanism of an embodiment of the rapid freezing apparatus of the present invention.

FIG. 10 is a diagram showing a schematic configuration of a conventional quick freezing apparatus.

[Explanation of symbols]

1: liquid nitrogen container, 2: body helium container, 3: liquid nitrogen, 4: liquid helium, 5: base cooling block, 6:
Metal block, 7: liquid helium transfer line, 8: decompression duct, 9: shutter, 10: sample insertion rod, 11: sample, 12: sample holder, 13: insertion guide tube, 14: heater, 15: liquid nitrogen container , 16: liquid helium container, 17: liquid nitrogen, 18: liquid helium, 1
9: liquid helium transfer tube, 20: liquid nitrogen filling port, 21: base cooling block, 22: metal block, 23: metal block rotary gear, 24: rotary drive mechanism, 25: rotary drive gear, 26: shutter, 2
7: Insulated tube, 28: Heater, 29: Sample insertion rod, 30:
Sample, 31: sample holder, 32: moving mechanism, 33: sample collection container, 34: push rod, 35: control device liquid nitrogen container, 36: cylinder, 37: compressed air, 38: solenoid valve, 39: moving mechanism Supporting part, 40: ascent position sensor,
41: descent position sensor, 42: intermediate position sensor, 4
3: Intermediate position sensor, 44: Ejector, 45: Liquid helium evaporating gas, 46: Temperature detector, 47: Rotating cylinder, 48: Direct position sensor, 49: Retreat position sensor, 50: Rotating cylinder, 51: Port switching Mechanism, 52: metal block exchange port, 53: sample insertion port, 54: outside air blocking lid, 55: light irradiation device, 5
6: Helium gas, 57: Metal block exchange rod, 58:
Body fluid nitrogen container, 59: body fluid nitrogen, 60: base cooling block, 61: temperature sensor, 62, 100: upper surface base, 200: housing.

(72) Inventor Noboru Moriya 2520 Akanuma, Hatoyama-cho, Hiki-gun, Saitama Prefecture Inside the Hitachi, Ltd.Basic Research Laboratories Co., Ltd. (72) Inventor Hisashi Otsuka 882 Ma, Hitachinaka City, Ibaraki Pref.Hitachi, Ltd.Measuring Instruments Division (72) Inventor Masaki Furuya 2520 Akanuma, Hatoyama-cho, Hiki-gun, Hiki-gun, Saitama Pref. Literature Jpn. Showa 57-755554 (JP, U) Jpn. Showa 57-75553 (JP, U) J. Sho 57-755552 (JP, U) (58) Fields investigated (Int. Cl. ⁷ , DB name) G01N 1/28 F25D 3/10 JICST file (JOIS)

Claims (6)

(57) [Claims] 1. A container for containing liquid helium, and the container
A metal block cooled to liquid helium temperature within
A heat insulating layer for accommodating a container for accommodating the liquid helium;
Housing and a cryogenic thermal conductivity that can hold the sample at the tip
Holding member having a high heat capacity and a small heat capacity, and an upper surface portion of the housing
And a predetermined sequence.
The test specimen moved according to the
Transfer after pressing the material onto the top surface of the metal block
A moving mechanism and the moving mechanism provided on the upper surface of the housing.
Opening that can pass and opens corresponding to the passage of the moving mechanism
Thermal shutter mechanism consisting of a shutter part
Means for heating the inside of the heat shut-off shutter mechanism
And moving the position of the moving mechanism at least to the ascending position and the descending position.
Position detecting means for detecting the position of the moving mechanism;
Therefore, when the shutter is opened,
Means for sucking outside air inside the
A rotation mechanism for updating the contact surface of the block with the sample,
A predetermined sequence operation corresponding to the detection signal of the position detection means
And a control device for performing the operation.
Freezing equipment . 2. The method according to claim 1, wherein the sample attached to the moving mechanism is
When pressed against a metal block, helium around the sample
Means for sucking gas gas to flow along the sample holder
The quick freezing apparatus according to claim 1, further comprising: 3. The moving mechanism according to a predetermined sequence.
When returning to the raised position after being lowered, the mobile device
Equipped with a sample collection means that can be moved to a position directly below the structure,
The moving mechanism is such that the sample collection means is located immediately below the moving mechanism.
After being moved to the
2. The rapid of claim 1 wherein the sequence is controlled to
Freezing equipment . 4. The apparatus according to claim 1, wherein the moving mechanism has a predetermined position.
After descending according to the sequence, return to the ascending position
Sequence control so that the camera can be rotated by a specified angle when
Quick freezing apparatus is Ru claim 1. 5. A sample is pressed against said metal block.
A light irradiating device for giving predetermined light to the sample before,
The irradiation is lowered by the moving mechanism according to a predetermined sequence.
2. The quick freezing apparatus according to claim 1, wherein the quick freezing apparatus is performed in the step of performing the freezing . 6. An upper surface of the nitrogen container is provided on an upper surface of the housing.
Open and pre-cool the metal block to liquid nitrogen in the nitrogen container
2. A base cooling block for immersion is provided.
The rapid freezing device as described .