JPS627830B2 - - Google Patents

Abstract

PURPOSE:To crush cells, etc. efficientyl, by setting vertically a cylindrical hood at the central part of a liquid bath for a cell crusher, floating a closed small container, in which cells are put, on the liquid level of the cylindrical hood, radiating ultrasonic wave from an ultrasonic oscillator to it so that the cells are crushed. CONSTITUTION:The bath 4 set at the top of a cell crusher is filled with the liquid W, and the cylindrical hood 6 having the open top 7 and bottom 8 is vertically fixed at the central part of the bath 4. The closed small container 13 putting cells to be crushed in it is fed to the liquid level L surrounded by the cylindrical hood 6 and floated. The oscillator 1 is driven, the magnetostrictive vibrators 2a, 2b, 2c, and 2d are vibrated through the excitation of the excited coil 13, and ultrasonic wave is radiated from the radiating face 17. Consequently, the closed small container 13 is rolled and moved on the liquid level L by the ultrasonic vibration, and the cells put in the small container 13 are crushed by the sound pressure of ultrasonic vibration while the cells are being subjected to stirring action.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to an apparatus for crushing cells, etc. using ultrasonic waves, and more specifically, an apparatus for crushing cells, etc. using ultrasonic waves suitable for crushing cell membranes of cells and separating intracellular organelles. Regarding.

Recently, in fields such as medical clinical testing and genetic engineering, methods of disrupting cell membranes and separating intracellular organelles have become popular in order to investigate the chemical composition and biological properties of each organ within the cell. It is being done. For example, it is widely used to advance biochemical analysis of the internal fluids of red blood cells and white blood cells in clinical tests.
For this purpose, an apparatus for disrupting cells using ultrasonic waves is used, and conventionally, an apparatus as illustrated in FIG. 5 has been frequently used. That is, oscillator 1' and vibrator 2'
For example, the horn or tip 3' is immersed in a beaker or test tube 4' containing a liquid containing red blood cells obtained by centrifugation from blood, and in that state, the oscillator 1 is activated. This device excites the vibrator 2' by the output of the oscillator 2' and applies ultrasonic vibration to the red blood cells in the container 4' through the horn 3', thereby causing cavitation phenomenon and the like to crush the cell membranes of the red blood cells. This conventional device fills a container 4' with a large amount of objects to be crushed, such as cells, and applies uniform ultrasonic vibration to the objects to be crushed from a horn 3', making it possible to crush a large amount at one time. On the other hand, in the case of the conventional type, the horn 3' remains inserted into the container 4', so it is necessary to keep the container 4' in an open state, and the non-sealed structure prevents toxic objects from being crushed. When processing, the objects to be crushed may be aerosolized by ultrasonic treatment and scattered outside, which is not desirable for inspectors and researchers.

Further, as mentioned above, since the structure is non-sealed and the water contained in the object to be crushed, such as cells, easily leaks out, the pH tends to change, and the desired crushing result cannot be obtained in many cases. For example, in the biochemical analysis of the internal fluids of red blood cells and white blood cells in clinical tests, leakage of water and changes in pH must be avoided as much as possible, but conventional methods have not adequately met these demands. In addition, the temperature of the liquid filled in the container has a great relationship with the propagation of ultrasonic vibrations and changes in the objects to be disrupted, such as cells, so it is desirable to appropriately control the liquid temperature during processing operations. However, in the case of the conventional type, the temperature of the liquid must be controlled by cooling the liquid from outside the container, which is difficult to control and difficult to manage. Therefore, for example, the lower the liquid temperature, the better the propagation of ultrasonic vibrations, and since most objects to be crushed are stored at low temperatures, if they are exposed to much higher temperatures during the crushing process, they will be exposed to heat. In these cases, the liquid temperature must be constantly cooled, but the conventional type cools the liquid from outside the container, so it could not be cooled as intended.

Furthermore, while the conventional apparatus is suitable for simultaneously crushing a large number of objects of the same type as described above, it is not suitable for efficiently crushing a variety of objects.

The present invention has been made in view of these points. It consists of a cylindrical hood suspended in the center with its lower part immersed in the liquid in the tank, a small container main body, and an airtight lid. This is an apparatus for disrupting cells, etc. using ultrasonic waves, which is equipped with a small container that can be suspended on the liquid surface, and its purpose is to provide a device that can crush objects to be disrupted in a closed manner, and has excellent safety. To provide a device that can suppress changes in objects to be crushed as much as possible, and to provide a device that can directly and extremely easily control the temperature of the liquid in a tank in which ultrasonic vibrations propagate.
Another object of the present invention is to provide a device suitable for efficiently crushing various types of objects to be crushed, and another object of the present invention is to provide a device that can efficiently utilize ultrasonic vibration energy.

The gist of the second invention of the present application is that, in the first invention, each of the magnetostrictive oscillators is arranged so that the ultrasonic waves radiated from the radiation surface at the bottom of the tank are aligned approximately on the center line of the cylindrical hood. An apparatus for disrupting cells, etc. using ultrasonic waves, characterized in that it is attached to a radiation surface while maintaining a predetermined angle so as to emit focused radiation to a small floating container; In addition to the above, the purpose is to further reduce the transmission loss and diffusion loss of ultrasonic energy in the liquid, so that objects to be crushed such as cells can be crushed more efficiently.

Next, an embodiment of the first invention of the present application will be described in detail with reference to the accompanying drawings FIGS. 1 to 4.

1 is a high frequency oscillator, 2a to 2d are vibrators such as ferrite magnetostrictive vibrators equipped with an excitation coil 3 driven by the high frequency oscillator 1, and 4 is a tank filled with liquid W. This is a conventionally known device. The present invention is characterized in the following points. That is, 6 is a cylindrical hood made of plastic or the like with upper and lower ends 7 and 8 open, and the cylindrical hood 6 is suspended in the center of the tank 4 with its lower part 9 immersed in the liquid W. The settings are maintained. In the case of the illustrated example, a cylindrical hood 6 is inserted into the central hooking hole 11 of the retaining plate 10 that is attached to the top of the tank 4.
An example is shown in which the hood 6 is held vertically by being inserted into a rack, and an example in which the lid 12 is placed on the top of the hood 6 is shown.

The present invention has a small sealed container 13 for floating on the liquid surface L surrounded by the hood 6. That is, a closed small container 13
consists of a small container main body 14 and a sealing stopper 15, and the small container main body 1
4, it is sealed using a sealing stopper 15, and then the object to be crushed A is placed inside the inner surface 16 of the small container main body 14.
The liquid is spread thinly and widely like a film, and then floated on the liquid level L in the cylindrical hood 6. In this case, the small container 13 is made of a material with light specific gravity such as plastic, and most of the interior is an air layer, so it floats on the liquid level L, and the length S of the small container 13 is equal to or smaller than the diameter of the hood 6. In comparison,
The small container 13 is designed to be small enough to move around freely within the hood 6.

Based on such a configuration, the crushing operation of this embodiment will be explained next.

First, the tank 4 is filled with liquid W. In this case, the amount of the liquid W is determined so that the liquid level L of the liquid W is located around the antinode of the ultrasonic vibration radiated from the ultrasonic radiation surface 17 of the tank 4. Next, a small amount of the object A to be crushed, such as cells, to be crushed is placed in the small container main body 14 constituting the small sealed container 13, and the container is sealed using the sealing stopper 15. Then, the closed small container 13 is lightly shaken to spread the crushed objects A inside it thinly and widely on the inner surface 16. Subsequently, the closed small container 13 is placed into the cylindrical hood 6. When poured, the small container 6 floats on the liquid surface L surrounded by the cylindrical hood 6. After this, the lid 12 is put on. Then drive the oscillator 1,
The magnetostrictive vibration 2 is caused to vibrate through the excitation of the excitation coil 3, and ultrasonic waves are emitted from the radiation surface 17. In this case, the thinner the plate thickness of the radiation surface 17 is, the larger the amplitude will be, and the larger the swing will be, so the radiation efficiency will be better, and it is preferable to set it to about 1.0 m/m to 0.8 m/m, for example.

Now, the ultrasonic vibrations radiated from the radiation surface 17 propagate through the liquid W and are radiated toward the closed small container 13 located inside the cylindrical hood 6 and floating above the liquid surface L. The object to be crushed A that is hermetically housed therein is crushed.

The crushing mechanism is as follows.
First, one of the crushing effects is that the ultrasonic vibration propagated in the liquid W is transmitted to the object A to be crushed in the closed small container 13, and the object A to be crushed is crushed by the sound pressure. In this case, the ultrasonic waves propagated in the liquid W are reflected at the air layer of the small sealed container 13, then reflected at the radiation surface 17 of the tank 4, etc., and are emitted again toward the object A to be crushed. By repeating this reflection, the object A to be crushed is successively crushed by the sound pressure. In the above, closed small container 1
When 3 receives the sound pressure of ultrasonic vibration, it rolls or moves on the liquid surface L and tries to escape to the surroundings where the ultrasonic vibration energy is weaker, that is, to the wall of the tank 4. However, since it is surrounded by the hood 6 in the central position, the closed small container 13
is always held in the central region where the ultrasonic vibration energy is strong, and the above-mentioned crushing action is effectively performed.

Another crushing effect is that instantaneous reduced pressure and compressive force alternately act on the liquid of the crushed object A due to the cavitation phenomenon that occurs in the liquid of the crushed object A due to ultrasonic vibration. The point is that each object A to be crushed is thereby crushed.

However, the present invention has the following features. That is, as described above, when the small sealed container 13 receives ultrasonic vibration, it rolls or moves on the liquid surface L surrounded by the hood 6. For example, as shown by the dashed line 18 in FIG. 2, or as shown by the arrow 19 in FIG.
The movements are small, rapid, and continuous. For this reason, the objects A to be crushed, which are thinly and widely adhered to the inner surface 16 of the small container main body 14 like a film, are subjected to constant stirring action. Therefore, the objects A to be crushed are sufficiently diffused within the inner surface 16, and each object A to be crushed in the liquid of the objects A to be crushed comes to be subjected to equally strong sound pressure of ultrasonic vibration.
That is, each object A to be crushed is inside the inner surface 16,
Since they move freely as the closed container rolls and moves, all of the individual objects A to be crushed are equally located at positions where they receive the sound pressure of strong ultrasonic vibrations, and some There is no bias where some things receive sound pressure while others receive weak sound pressure. Moreover, the diffusion due to the stirring action described above is accomplished within a short period of time. These crush the particles very efficiently.

Next, the second invention of the present application will be explained in detail.

That is, the magnetostrictive vibrators 2a, 2b, 2c, 2d
Each of the ultrasonic waves emitted from the ultrasonic emitting surface 17 at the bottom of the tank 4 is formed into a radiation surface so that the ultrasonic waves can be focused and radiated to the small sealed container 13 floating approximately on the center line C of the cylindrical hood 6. 17 at a predetermined angle a,
b.

In this case, the ultrasonic vibrations radiated from the radiation surface 17 due to the vibrations of the magnetostrictive oscillators 2a to 2d rise above the liquid level L in the hood 6 as indicated by numbers 20, 21, 22, and 23. It focuses toward the closed container 13 floating in the air. Therefore, diffusion loss and transmission loss occurring during the propagation process of ultrasonic vibrations are suppressed as much as possible, and effective vibration energy is applied to the object A to be crushed within the closed small container 13. On the other hand, the small container 13 that receives the focused vibrational energy rolls and moves more actively and tries to escape to the surroundings where the energy is weaker, but as described above, it is held in the central position by the central cylindrical hood 6. Therefore, the effect of the above-mentioned focused radiation can be utilized as is. Therefore, according to this second invention,
More efficient crushing can be achieved without loss of ultrasonic vibration energy.

In both the first invention and the second invention, the object A to be crushed, such as cells, is placed in the closed small container 13.
Since the object A is crushed in a sealed state,
There is no risk of it turning into aerosol and scattering outside. Therefore, it is extremely safe for researchers and inspectors, and the desired crushing results can be obtained since there is no leakage of water or any accompanying change in pH of the object A to be crushed. . Furthermore, since the object A to be crushed is sealed in the small closed container 13, the liquid W in the tank 4 can be directly cooled. That is, if you want to lower the temperature of the liquid, you can directly adjust the temperature of the liquid by directly injecting lower-temperature cooling water into the tank. Since the liquid temperature can be easily adjusted in this way, it is possible to improve the propagation of ultrasonic vibrations and to prevent thermal changes in the object A itself to be crushed as much as possible. Also, if you seal a number of different types of objects to be crushed in a number of containers and put them into the hood one after another, or if you put as many sealed small containers as possible into the hood, you can crush a number of objects in a single hour. It is capable of efficiently crushing objects to be crushed, such as seeds, and is effective in crushing a wide variety of small quantities.

As shown in FIG. 4, a rod 24 such as a metal rod such as aluminum or an acrylic rod is housed in the small airtight container 13, and the liquid L is
It may also be crushed by floating on top. In this case, since vibration of the rod 24 is induced, vibration energy is also applied from the rod 24 to the object to be crushed, and the crushing action becomes more multifaceted.

As detailed above, the first invention of the present application is a device comprising a magnetostrictive vibrator that generates ultrasonic waves and a tank filled with liquid, in which the lower part is immersed in the liquid in the tank in the center of the tank. It consists of a cylindrical hood suspended in a suspended state, a small container main body, and an airtight lid. Since this is a device for crushing cells, etc., it is possible to crush objects to be crushed in a closed manner, providing a device with excellent safety, as well as suppressing changes in the object to be crushed itself, and being able to obtain the desired crushing results. and
In addition, the liquid temperature in the tank can be directly adjusted and easily managed, and various problems caused by rising liquid temperature can be corrected.Furthermore, various types of objects to be crushed can be efficiently crushed. The object to be crushed can be crushed by efficiently utilizing the sonic vibration energy. Further, in a second invention of the present application, in the first invention, each of the magnetostrictive vibrators is a small container in which ultrasonic waves emitted from a radiation surface on the bottom of the tank float approximately on the center line of the cylindrical hood. Since this is an ultrasonic cell disruption device that is mounted at a predetermined angle with respect to the radiation surface so as to emit radiation in a focused manner, in addition to the above advantages,
This reduces the transmission loss and diffusion loss of ultrasonic energy in the liquid, making it possible to more efficiently crush objects such as cells.

[Brief explanation of the drawing]

The accompanying drawings show an embodiment of the present invention, and FIG. 1 is a longitudinal side view, FIG. 2 is a sectional view taken along the line X--X in FIG. 1, and FIG. 3 is a sectional view showing the relationship between the hood and the closed container. is a vertical cross-sectional view showing the relationship between the hood and the closed container;
FIG. 4 is a diagram showing an embodiment in which an aluminum rod is housed in a closed small container, and FIG. 5 is a diagram of a conventional example. 2a to 2d... magnetostrictive vibrator, 4... tank, 6...
Cylindrical hood, 9... Lower portion of cylindrical hood, 13...
Sealed small container, 14... Sealed small container body, 15...
Sealing stopper, 16...inner surface, 17...ultrasonic radiation surface,
W...liquid, L...liquid level in the hood 6.

Claims (1)

[Claims] 1. In a device consisting of a magnetostrictive vibrator that generates ultrasonic waves and a tank filled with liquid; A device for disrupting cells, etc. using ultrasonic waves, comprising: a cylindrical hood; a small hermetically sealed container consisting of a small container main body and a sealed lid, in which an object to be disrupted is hermetically housed and floated on the liquid surface within the cylindrical hood; . 2 In the invention described in claim 1,
Each of the magnetostrictive oscillators is arranged at a predetermined angle with respect to the radiation surface so that the ultrasonic waves emitted from the radiation surface on the bottom of the tank are focused and radiated to the small container floating approximately on the center line of the cylindrical hood. An apparatus for disrupting cells, etc. using ultrasonic waves, which is characterized in that it is installed while maintaining the installation angle.