JPH06328002A - Precipitation classifying method and apparatus therefor - Google Patents

Abstract

(57) [Summary] [Purpose] It is an object of the present invention to provide a sedimentation-type classification method and apparatus capable of classifying a large number of classified substances having various groups such as cell clumps at each growth stage into a large number. . [Structure] Cylindrical classification tank 4 arranged axially horizontally
A plurality of partition portions 6 are arranged in the lower axial direction of. The first inclined conical pipe line 2 whose upper portion is kept at a constant height and whose lower portion rises obliquely outward and whose front end portion is open is connected to one opening end of the classification tank 4. At the other opening end of the classification tank 4, a second inclined conical pipe line 10 whose lower part is kept at a constant height and whose upper part is inclined downward toward the outside and whose front end is open is connected. . Tip 2 of the first oblique conical conduit 2
The mixed liquid of the object to be classified flows in from a, the liquid flows out from the tip portion 10a of the second oblique conical conduit 10, and the object to be classified is classified into the partition part 6 due to the difference in the terminal speed of the falling of the object to be classified. I decided to do it.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a settling type classification method and apparatus, and more particularly, to classify many kinds of classified materials in large quantities by utilizing the difference in terminal speed of classified objects mixed in a liquid. The present invention relates to a sedimentation-type classification method and device, particularly to a sedimentation-type classification method and device suitable for classification of cell aggregates and the like.

[0002]

2. Description of the Related Art Conventionally, as a method for selecting minute objects in the form of powder or granules, a method of sieving using a sieve, a method of using a centrifuge, or a method utilizing a difference in sedimentation velocity in a fluid is used. There has been considered a method using a classifying device for classifying the A. into groups having different sizes or densities.

For example, the inventions described in JP-B-52-47715 and JP-A-2-247541 enable precise classification using a centrifugal separator. However, since this method is a batch method, continuous selection cannot be performed, and in the case of classifying cell aggregates, there is a risk that contaminants and impurities will be contaminated when the cells are put into and taken out from the centrifuge.

The following classifying device has also been proposed. That is, the device described in Japanese Utility Model Laid-Open No. 63-99259 is a device for free fall of objects in a classification tank, and a take-up belt installed at the bottom of the tank to take out the objects in order from the objects falling on the belt and perform classification. is there. However, continuous classification is not possible with this apparatus, and it takes time to take out the classified objects from the belt, so that it is not possible to classify a large amount. Further, there is a possibility that impurities such as oil and metal powder generated from the winding portion of the belt may be mixed in, which is not suitable for classifying cell aggregates.

The inventions described in JP-B-3-99, JP-A-59-196760 and JP-A-3-178352 are those which do not cause such a problem. Special Fairness 3-
In the invention described in No. 99, a dispersion liquid in which an object group including an object to be classified is dispersed in a fluid is put into a container equipped with a piston whose bottom part can slide up and down, and the bottom part of the container is pushed up at a constant speed. As a result, an upward flow that prevents turbulent flow is created, and particles having a sedimentation velocity lower than the pushing-up velocity are caused to overflow from the upper part of the container, so that highly accurate classification can be performed.

Further, in the inventions disclosed in JP-A-59-196760 and JP-A-3-178352, the density of the fluid is adjusted so that an object having a density higher than the adjusted density is settled, and a small density is set. Levitate an object with
It is classified into two groups. Furthermore, Japanese Patent Publication No. 57-5
No. 582 is a method in which a suspension containing microscopic objects is allowed to stand in a tank, is allowed to settle, is extracted by a suspension withdrawal tube, and is classified. In all of the above, the material to be classified is divided into two groups.

The invention described in Japanese Examined Patent Publication No. Sho 53-16933 is a combination of upright cylinders and expanding cylinders which are alternately combined, the structure of which prevents turbulence and enables classification into multiple groups. Is.

Furthermore, the device described in Japanese Utility Model Application Laid-Open No. 58-166882 is a classifier for powders and granules, in which the powders and granules to be classified are supplied from above to a flowing gas, By using the descent speed difference of, the classification is performed according to the difference of the falling position.

[0009]

However, most of these methods only divide the object into two groups and cannot classify the cell mass into multiple groups according to the growth stage, and turbulent flow in the tube. Therefore, classification accuracy suitable for objects such as cell clusters cannot be expected. In addition, Japanese Patent Publication Sho 53-1
The invention described in No. 6933 has a narrow entrance of an object recovery port and cannot recover an object having a sedimentation velocity equal to the in-pipe flow velocity in the expanding cylindrical body part, so that the recovery efficiency of the object as a whole is poor and a large amount of There is a problem that you cannot classify. Further, in the device described in Japanese Utility Model Publication No. 58-166882, since the wind force, gravity, and frictional resistance applied to the object to be classified do not balance during classification, the object falls in a curved line, and the falling trajectory becomes unstable. The falling position cannot be accurately grasped and the classification accuracy is poor. Also, in the case of culturing / growing in a culture medium like a cell mass, it is necessary to perform classification / selection in a liquid,
Classification in a gas like this device is not suitable.

It is an object of the present invention to provide a sedimentation type classifying apparatus capable of classifying a large number of classified substances having various groups such as cell clumps at each growth stage into a large number.

[0011]

A settling type classification method according to the present invention is a dropping position based on a terminal velocity of a classified substance in which a liquid mixed with the classified substance is caused to flow at a constant velocity and settles in the liquid. The feature is that the objects to be classified are classified according to the difference of.

Further, in the sedimentation type classification method according to the present invention, the object to be classified is dropped in the liquid flowing at a constant speed, and the classified object is classified by the difference in the falling position based on the terminal velocity of the object to be settled in the liquid. It is characterized by classifying things.

Further, the sedimentation type classifying apparatus according to the present invention has a cylindrical classifying tank whose axial direction is horizontally arranged and a lower axial direction of the classifying tank, which is arranged by opening a valve to obtain another type. A plurality of partition parts communicating with the device and one end of the opening of the classification tank are connected, the upper part is kept at a constant height in a side view, the lower part rises obliquely outward, and the tip part is It is connected to the opened first oblique conical pipe line and the other opening end of the classification tank, the lower part is kept at a constant height in a side view, and the upper part is inclined downward toward the outside. A second oblique conical conduit having an open tip, and the mixed liquid of the material to be classified is caused to flow into the second oblique conical conduit from the distal end of the first oblique conical conduit. The feature is that the liquid is made to flow out from the tip.

Further, the settling type classifying apparatus according to the present invention has a cylindrical classifying tank whose axial direction is arranged horizontally and a lower axial direction of the classifying tank, and is opened by opening a valve. A plurality of partition parts communicating with the device, a first conical conduit connected to one opening end of the classification tank, narrowing outward, and having an open tip, and the other of the classification tanks A second conical pipe line connected to the end of the opening of the container, narrowing outward, and having an open tip, and a rectifying grid disposed near the end of the one opening of the classification tank. , Letting the liquid flow in from the tip of the first conical conduit and letting the liquid flow out from the tip of the second conical conduit,
It is characterized in that an object to be classified is introduced through an introduction port provided in an upper portion of the classification tank near the end of the one opening.

Furthermore, the object to be classified by the sedimentation type classification method and apparatus according to the present invention is characterized in that it is a cell mass.

[0016]

As described above, according to the present invention, the liquid mixed with the substance to be classified is caused to flow at a constant velocity, or the substance to be classified is dropped into the liquid flowing at a constant velocity. ，
The buoyancy and the frictional resistance with the liquid balance each other, and the liquid settles at a constant speed. This speed is called the terminal speed, and it depends on the density, mass, volume, shape, etc. of the material to be classified. The object to be classified falls along a linear trajectory obliquely downward due to the terminal velocity and the flow velocity of the liquid. The objects to be classified are classified according to the difference in density, mass, etc. due to the difference in the falling position.

Further, according to the present invention, when the mixed liquid of the substance to be classified is caused to flow in from the tip of the first oblique conical conduit and the liquid is caused to flow out from the tip of the second oblique conical conduit, The substances to be classified mixed in the liquid sequentially enter the partition section arranged in the lower axial direction of the classification tank according to the difference in the terminal speed. By opening the valves connected to these partitions,
The objects to be classified that have been classified into the partition section are carried to another device.

Furthermore, the liquid is allowed to flow in from the tip of the first conical conduit and the liquid is allowed to flow from the tip of the second conical conduit, and the one opening of the classification tank is also used. When the material to be classified is introduced from the inlet provided at the upper part near the end, the material to be classified is settled at a constant speed in the liquid with less turbulence by the rectifying grid, and the partition part arranged in the lower axial direction of the classification tank. Go into order. By opening the valves connected to these partition parts, the objects to be classified that have been classified into the partition parts are carried to another device.

[0019]

Embodiments of the present invention will be described below with reference to the drawings. Before that, the principle of the sedimentation classifier using gravity will be described. When an object freely falls in a stationary liquid,
Gravity, buoyancy, and frictional resistance with fluid balance the object, and eventually the object will fall at a constant velocity. This velocity is called the terminal velocity, and it depends on the properties such as the mass, volume, density, and shape of the object. This operation is constant in the horizontal direction,
When performed in a pipe extending in the horizontal direction where the flow velocity is the same at any place in the flow path, two objects with different terminal velocities are
As shown in, there is a difference in the horizontal movement distance from the drop start position to the drop end position. Classifying an object by the difference in the horizontal movement distance generated at this time is a sedimentation type classification method using gravity.

FIG. 1 shows a first embodiment of the sedimentation type classifying apparatus according to the present invention. In the figure, reference numeral 4 is a cylindrical classification tank having a smooth inner side, which is arranged horizontally in the axial direction, and one end of the opening of the classification tank 4 is kept at a constant height in a side view from the top. , The first oblique conical conduit 2 whose lower portion rises obliquely outward and whose tip 2a is open
Are attached with screws using the flange 5. The first inclined conical conduit 2 gradually expands toward the classification tank 4 to the extent that the liquid does not become a turbulent flow when viewed from the tip 2a.

The tip portion 2a of the first oblique conical conduit portion 2
1, a pipe 1 is detachably attached using a union 3, and the pipe 1 is connected with a pump or the like for feeding a suspension containing minute objects of cell mass to be classified. At the bottom of the classification tank 4, cylindrical partition parts 6 (6a, 6b, ...) that narrow downward are arranged in the axial direction of the classification tank 4, and at the bottom of these partition parts 6 there are half unions. 7 are connected, and tubes 8 made of silicon or the like can be easily attached and detached. In addition,
The tube 8 made of silicon or the like is opened and closed by the pinch cock 9.

At the other end of the opening of the classification tank 4, the inside is smooth to the extent that turbulent flow does not occur, the upper part rises obliquely outward, and the lower part is kept at a constant height in side view. The second oblique conical conduit 10 with the tip end 10a opened
However, it is screwed 11 using the flange 11.
A pipe 13 is detachably attached to a tip portion 10a of the second oblique conical conduit 10 by a union 12.

Next, the operation of this device will be described.
When the minute object to be classified is a cell mass and the liquid is a liquid having a density of 1 g / cm ³ such as a culture solution, the cell mass flowing from the culture tank in the pipe 1 at a constant flow rate with the liquid is
First, the first inclined conical conduit 2 is entered. Liquid has a density of 1g /
Since it is cm ³ , and the density of the cell mass is slightly higher than 1 g / cm ³ , in the liquid of this example, gravity, buoyancy, and frictional resistance applied to the cell mass are immediately balanced, and if we focus only on the vertical movement, , Immediately after the fall starts, it falls at a constant speed.
The velocity of the cell mass at this time is called the terminal velocity.

In the first oblique conical conduit 2, the cell mass falls in the vertical direction at a constant velocity, but since the conduit gradually expands, the liquid flow velocity gradually decreases in the horizontal direction. The mass falls in a parabola as shown. When the cell mass enters the classification tank 4 through the first oblique conical conduit 2, the classification tank 4 flows horizontally at a constant and equal flow rate at any point in the tank.
The cell clumps fall in a diagonal straight line as shown in the figure,
The particles enter the partition section 6 at the bottom of the classification tank 4, pass through the silicon tube 8, and are transported to the next step. At this time,
The larger the cell mass with the faster terminal velocity and the larger shape, the closer to the partition 6a on the left side in the figure, and the smaller the cell velocity with the lower terminal speed and the smaller the shape, the closer to the partition 6d on the right side. It can be classified according to the growth stage. The cell mass having a slow terminal velocity that does not enter the partition 6d is returned together with the liquid to the culture tank through the second oblique conical conduit 10 and the pipe 13, and is cultured again.

From the above, by using the sedimentation type classification method and apparatus of the embodiment of the present invention, it is possible to continuously classify a large amount of minute objects into a large number of groups. In addition, since classification is performed in a pipeline isolated from the outside, it is not contaminated with various bacteria and impurities, and since the device can be disassembled, the device can be easily washed and sterilized. It is also effective for objects. Also, because liquid is used as the fluid, the wind force, gravity, and frictional resistance applied to the object to be classified like gas are not balanced during classification, the object falls in a curved line, and the fall trajectory becomes unstable. There is no drawback that the falling position cannot be accurately grasped and the classification accuracy is poor. Further, in the case of culturing / growing in a culture solution such as a cell mass, classification / selection also needs to be performed in the liquid, and thus the apparatus according to this example is suitable for classification.

Next, a second embodiment of the present invention will be described. The same or corresponding parts as those described in the first embodiment are designated by the same reference numerals. FIG. 2 shows a second embodiment of the sedimentation type powder flow material classifying apparatus according to the present invention. In the figure, 4 is a cylindrical classification tank in which the axial direction is horizontally arranged and the inside is smooth. At one end of the opening of the classification tank 4, the classification tank 4 is tapered outward and has a tip 15a. An open first conical conduit 15 is attached with screws using the flange 5. This first oblique conical conduit 15
Is gradually expanded toward the classification tank 4 so that the liquid does not become a turbulent flow when viewed from the tip portion 15a.

At the other end of the opening of the classification tank 4,
The second taper toward the outside, with the tip 17a opened
The conical conduit 17 is attached with screws using the flange 5. This second oblique conical conduit 17 also has a tip portion 1.
When viewed from 7a, the liquid gradually expands toward the classification tank 4 to the extent that the liquid does not become a turbulent flow.

The tip portion 15 of the first oblique conical conduit 15
A pipe 1 is detachably attached to a by using a union 3, and a pipe 13 is connected to a tip portion 17a of the second oblique conical conduit 17 by a union 12. Although the circuit 13 is connected to the pipe 1 after making one round, a drain 18, a pump 19, a flow rate adjusting valve 20, and a flow meter 21 are sequentially connected in the line of the pipe 13 → the pipe 1.

A rectifying grid 16 is arranged near the end of the one opening of the classifying tank 4, and the rectifying grid 1 is provided.
6 is made of an extremely thin material in a lattice shape, and is attached by a detachable fitting type. The rectifying grid 16 rectifies the liquid that has flowed in from the first conical conduit 15 so that it does not become a turbulent flow. Also,
A charging hole 4a for charging a material to be classified is provided in the upper part of the classification tank 4 in the vicinity of the one end of the opening, and a pipe 27 extending vertically is provided in the charging hole 4a.
8 are used for connection. A ball valve (ON / OFF valve) 29 is arranged in the middle of the pipe 27, and a container (not shown) containing a suspension containing minute objects such as cell clusters for classification is connected to the upper end. Has been done. FIG. 4 shows a specific shape of the present device, (a) is a plan view and (b) is a side view, and the reference numerals shown in FIG.

Further, in the lower part of the classification tank 4, as in the case of the first embodiment, a plurality of cylindrical partition parts 6 (6, which are narrowed downward, are formed.
a), 6b, ...) are provided, and half unions 7 are connected to the lowermost portions of the partition portions 6, respectively, so that tubes 8 made of silicon or the like can be easily attached and detached. The tube 8 made of silicon or the like is opened and closed by the pinch cock 9. The other end of the tube 12 made of silicon or the like is connected to each subsequent step.

Next, the operation of this device will be described.
The inside of the device is filled with a liquid, and the liquid is circulated in the device by a pump 19. The flow direction is from left to right in the figure in the classification tank 4. Also, the flow velocity in the classification tank 4 is
It is controlled by the flow rate adjusting valve 20 downstream of 9, and its speed can be known by measuring the value of the flow meter 21 downstream of the flow rate adjusting valve 20. Further, in order to prevent the liquid in the classification tank 4 from becoming a turbulent flow due to the sudden expansion and contraction of the conduit, the first conical conduit 15 and the second conical conduit 15 are provided on the left and right of the classification tank 4.
The conical pipe line 17 is connected, and the flow in the classification tank 4 is maintained at a constant and equal flow rate in the horizontal direction at any point by the rectifying grid 16. The parts are removable so that the device can be easily cleaned and sterilized.

As an example, a minute object for classification will be described as a cell mass and a liquid as a culture solution. First, the cell mass is pipe 27
Is dropped and supplied to the classification tank 4. At this time, the relationship between the density of the liquid culture medium and the cell mass is that the density of the cell mass is slightly higher than that of the culture liquid, so that the cell mass falling in the liquid has gravity and buoyancy applied to the cell mass immediately after falling. The frictional resistance balances out and it immediately drops at a constant speed. The speed at this time is called the terminal speed.

The cell clumps fall through the pipe 27 and the classification tank 4
When it falls inside, it reaches the terminal speed, and thereafter it drops at a constant speed. Since the liquid flows horizontally in the classifying tank 4 at a constant flow velocity, the cell clumps entering the classifying tank 4 fall in a slanting downward straight line as shown in the figure, and the partitioning section at the lower part of the classifying tank 4 falls. 6 (6a, 6b, ...) Enter and be classified. At this time, a larger cell mass having a shape with a higher terminal speed enters the left partition 6a, and a smaller cell mass having a shape with a lower terminal speed enters the right partition 6b. In this way, the cell mass can be classified and sorted according to the size of the shape, that is, the growth stage. The cell mass that has entered each partition 6 is transferred to the next step through a tube 8 made of silicon or the like.

As shown in FIG. 4, the inside diameter of the classification tank 4 is R, the flow velocity of the liquid is V, the vertical downward velocity U of the minute object to be classified, the horizontal moving distance of the minute object is X, and the minute object is Let t be the time it takes for the object to fall (time to reach the horizontal movement distance X), since a minute object moves at a constant velocity in both horizontal and vertical directions, t = R / U ..・ T = X / V ...

From the above, the horizontal movement distance X is X = RV / U .... Therefore, the horizontal moving distance of the minute object can be easily obtained by calculation if the diameter and flow velocity inside the classification tank 4 and the terminal velocity of the object are known. Therefore, if the terminal speed of the object to be classified is known, the diameter of the classification tank 4, the length in the horizontal direction, the flow velocity, and the position of the partition plate for providing the partition 6 can be set by calculation. it can.

Next, an example of an experiment conducted using this apparatus will be described with reference to FIG. (Experimental example) A carrot somatic embryo having a major axis length of 1 to 4 mm (a cell mass having a part to be a future sprout and a root produced by cutting out a part of a carrot living body and culturing) and having a major axis length of 1 to 2 m
It is assumed that the carrot somatic embryos of 3 groups of m, 2 to 3 mm, and 3 to 4 mm are classified. To carry out this classification, the diameter inside the classification tank 4 is 300 mm, the flow rate is 3 mm / sec, and the liquid is 0.1
% Aqueous solution of sodium alginate. First, major axis length 1
Terminal velocities of carrot somatic embryos of ~ 2 mm, 2-3 mm, 3-4 mm average 1.772, 2.443, 3.503 mm /, respectively.
Since it is sec, the horizontal movement distances are 507.9, 368.4, and 256.9 mm from the above equation, respectively. From this,
The position of the partition plate for providing the partition portion 6 is set to the center of 256.9 to 368.4 mm from the center of the pipe 27 and 368.4 to 507.9.
It was provided at the center of mm. As a result, 3
A group of somatic embryos could be obtained.

[0037]

As described above, according to the present invention,
It is possible to classify an object to be classified such as a cell clump that has various groups at each growth stage into a large number of groups and prevent contamination with impurities such as various bacteria.

[Brief description of drawings]

FIG. 1 is a side view of a first embodiment of the present invention.

FIG. 2 is a side view of the second embodiment of the present invention.

FIG. 3 is a diagram illustrating a classification principle by sedimentation.

4A and 4B are views showing a specific external appearance of a sedimentation type classification device according to a second embodiment, FIG. 4A being a plan view and FIG. 4B being a side view.

5 is a diagram illustrating the principle of the second embodiment shown in FIG.

FIG. 6 is a diagram for explaining an experimental example.

[Explanation of symbols]

 1 Pipe 2 1st Inclined Conical Pipe 3 Union 4 Classification Tank 4a Input Hole 5 Flange 6 Partition 7 Half Union 8 Silicon Tube 9 Pinch Cock 10 Second Oblique Conical Pipe 11 Flange 12 Union 13 Pipe 15 1st Conical conduit 16 Straightening grid 17 Second conical conduit

Claims (5)

[Claims] 1. A method in which a liquid mixed with an object to be classified is caused to flow at a constant velocity and the object to be classified is classified according to a difference in a drop position based on a terminal velocity of the object to be classified that is settling in the liquid. A characteristic sedimentation type classification method. 2. The object to be classified is dropped in a liquid flowing at a constant velocity, and the object to be classified is classified according to the difference in the drop position based on the terminal velocity of the object to be settled in the liquid. Settling type classification method. 3. A cylindrical classification tank whose axial direction is arranged horizontally, and a plurality of partition portions which are arranged in the lower axial direction of the classification tank and which communicate with other devices by opening a valve, A first oblique conical conduit which is connected to one opening end of the classification tank, has an upper part kept at a constant height in a side view, and has a lower part rising outward obliquely and having an open end. And a second oblique cone connected to the other opening end of the classification tank, the lower part of which is kept at a constant height in a side view, the upper part of which falls obliquely outward, and the end part of which is open. And a liquid pipe, and a liquid mixed with the substance to be classified is caused to flow in from a tip portion of the first oblique conical pipe passage, and a liquid is caused to flow out from a tip portion of the second oblique conical pipe passage. A sedimentation classifier characterized by the following. 4. A cylindrical classification tank whose axial direction is arranged horizontally, and a plurality of partition sections which are arranged in the lower axial direction of the classification tank and which communicate with other devices by opening a valve, A first conical pipe line connected to one opening end of the classification tank, narrowing outward, and having an open tip, and connected to the other opening end of the classification tank A second conical pipe line that narrows toward the end and has an open tip, and a rectifying grid that is arranged near the one opening end of the classification tank, and the first conical pipe line of the first conical pipe line is provided. The liquid is made to flow in from the tip portion, and the liquid is made to flow out from the tip portion of the second conical conduit, and classification is performed from an input port provided in the upper portion of the classification tank near the one end of the opening. A sedimentation type classifier characterized in that it is adapted to throw in a substance. 5. The sedimentation type classification method and apparatus according to claim 1, 2, 3 or 4, wherein the material to be classified is a cell mass.