JP2014136213A - Rotor of centrifugal separator, and centrifugal separator - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a rotor of a centrifugal separator for a microplate capable of shortening the acceleration-deceleration time by size-weight reduction, and capable of eliminating the difference between a sample temperature and a centrifugal machine preset temperature.SOLUTION: A rotor of the centrifugal separator is provided for centrifugally separating a sample injected into a microtube aggregate. The rotor of the centrifugal separator comprises a bottom part fixed to a rotary shaft of the rotor and a rotor sidewall of extending upward from this bottom part, and in the rotor of the centrifugal separator, an inner peripheral part of the rotor sidewall is formed with a microplate holding part composed of an aggregate of a plurality of through-holes capable of inserting tubes of the microtube aggregate, and the microplate holding part is formed in a dug recess having a flat bottom part formed in an inner peripheral part of the rotor sidewall.

Description

  The present invention relates to a centrifuge rotor used in the fields of medicine, pharmacy, genetic engineering, etc., and in particular, using a microplate, applies centrifugal acceleration to a sample in a tube in which a solvent solution and fine particles are mixed. The present invention relates to a rotor of a centrifuge for separating a solvent liquid and fine particles in a sample and a centrifuge using the rotor.

As a sample box for processing a large number of samples at the time of centrifugation, a microplate comprising a flat plate and a large number of wells formed in the flat plate and receiving the sample is used.
Recently, a microplate-like tube assembly (PCR microplate) in which a thin flat plate and a large number of microtube assemblies for receiving a sample project from an opening formed in the flat plate are integrally provided. Well used.
The microtube has a closed bottom and the number of microtubes is 8,24,96,384,1536. Each tube has a flat bottom, a U-shaped bottom, and a V-shaped bottom. Depending on the purpose of use, the bottom of the tube can be selected from a shallow bottom to a deep bottom (deep well plate). The material is made of polystyrene, polypropylene, polyethylene or the like according to the application, and by using such a microplate, a large number of separation processes can be performed simultaneously.

  For example, the test accuracy is improved by amplifying DNA in a minute sample by a polymerase chain reaction used for DNA research or criminal investigation and amplifying it for the determination of subsequent research or criminal investigation.

  When centrifuging the sample injected into the microtube, centrifugal acceleration is applied to the microplate. For example, when a deep well plate is used, a box type sample holder in which a support is fitted to the lower side of the deep well plate is used, but this microplate is generally made of polystyrene or polypropylene. Therefore, it was mechanically fragile and could not withstand the centrifugal acceleration force during the centrifugal operation, possibly causing damage or deformation.

Therefore, in order to protect the deep well plate from this breakage and deformation, an adapter closely attached to the shape of the box-type sample holder is attached.
This box type sample holder is disclosed in Japanese Patent No. 4482658. As shown in FIG. 14, the box-shaped sample holder 100 and the adapter 110 are polymerized in a state of being fitted to each other, and are fitted into a cut-in insertion portion 400 formed on the inner wall surface side of the rotor. The adapter 110 is formed with a plurality of movement preventing pieces 110a erected from the bottom of the adapter, and the movement preventing pieces 110a are fitted between a plurality of sample storage tubes 140 arranged in a box-shaped sample holder. Thus, each sample storage tube in the box-shaped sample holder is fixed to the adapter 110 so as not to be deformed. The rotor has a hollow inverted frustoconical shape (a shape in which the opening diameter increases toward the upper side of the rotor), and the adapter 500 is formed by the centrifugal acceleration force at the time of centrifugation by the detent 500 formed in the recessed insertion portion 400. This prevents the box-shaped sample holder 100 fixed in step (5) from moving upward along the inner wall surface of the rotor.

  On the other hand, when using a microplate other than the box-type sample holder, each sample storage tube is similarly fixed by an adapter so as not to be damaged or deformed by the centrifugal acceleration force. Then, the microplate fixed by the adapter is fixed so as not to move upward along the inner wall surface of the rotor, like the box-type sample holder.

Japanese Patent No. 4482658

  In the above conventional centrifuge, especially when used at a rotational speed of 3,000 rpm or more, the microplate constituting the box-type sample holder is deformed unless a dedicated adapter suitable for the box-type sample holder is attached. -Centrifugal operation is not possible due to possible damage. Therefore, when adapters corresponding to various types of box-type data holders are mounted, the box-type data holders may interfere with each other, and therefore the outer diameter of the rotor becomes large, and a generally-used small centrifuge Has the disadvantage that it cannot be used.

  In addition, since the adapter has a suitable thickness to withstand centrifugal force, it is difficult to reduce the weight.Therefore, it takes time to accelerate and decelerate the rotor, and there is a limit to centrifuging samples quickly and in large quantities. there were. Furthermore, if the capacity of the microtube assembly is increased in order to centrifuge a large amount of the sample, a metal adapter is required, and there is a problem that it takes a long time to accelerate and decelerate the rotor due to an increase in weight.

For this reason, many adapters are molded of plastics for weight reduction.
However, when cooling the sample temperature heated to a predetermined temperature by centrifugal acceleration this time, the plastic adapter has a low thermal conductivity, so that there remains a problem that the cooling efficiency of the sample is poor.

  In the present invention, the microplate is directly fixed to the rotor without using an adapter, so that the centrifuge rotor can be reduced in size and weight, the cooling time of the sample temperature in the microtube assembly can be shortened, and the set temperature can be reduced. An object of the present invention is to provide a centrifuge rotor that can eliminate the difference.

  The present invention has been made to solve the above-described problems. A microplate in which a flat plate and a microtube assembly protruding from the flat plate and into which a sample is injected are integrally provided with a rotor of a centrifuge. In the centrifuge rotor for centrifuging the sample injected into the microtube assembly by holding and rotating the centrifuge, the centrifuge rotor has a bottom fixed to the rotating shaft of the rotor and an upper portion from the bottom. And a microplate holding part formed of an assembly of a plurality of through holes into which the tube of the microtube assembly can be inserted is formed on the inner peripheral part of the rotor side wall. And

  According to the present invention, the microplate holding portion formed of an assembly of a plurality of through holes into which the tube of the microtube assembly can be inserted is formed on the inner peripheral portion of the rotor side wall. It can be directly held on the rotor inner wall without attaching an adapter.

This eliminates the need for an adapter, thus reducing the size and weight of the centrifuge rotor and shortening the acceleration / deceleration time during centrifugal rotation. Furthermore, since the microplate can be directly held in the microplate holding part, the tube of the microtube assembly into which the sample is injected is exposed in the centrifuge chamber from the through hole formed in the holding part, The temperature in the temperature-controlled chamber can be directly transmitted without passing through the rotor side wall. Therefore, since the cooling time of the sample temperature in the microtube assembly can be shortened, the sample can be centrifuged in a large amount more quickly.
Moreover, the difference between the sample temperature in the microtube assembly and the set temperature can be eliminated.

In the above invention, it is desirable that the microplate holding portion is formed in a recessed portion having a flat bottom formed on the inner periphery of the rotor side wall. By configuring in this way, when the microtube assembly is inserted into the through-hole, the flat plate of the microplate is in close contact with the flat bottom of the dug recess, thus preventing deformation of the microplate during centrifugal acceleration. can do.
In particular, it is suitably applied to a rotor side wall having a curved surface.

In the above invention, it is desirable to provide a notch that communicates from the side wall of the digging recess to the bottom of the digging recess.
With this configuration, when taking out the microplate that is in close contact with the microplate holding part by centrifugal acceleration, a fingertip is inserted into the notch groove, and the finger is applied to the back of the flat plate of the microplate. Can be easily taken out without applying a load. In addition, the efficiency of the operation of detaching the microplate from the rotor can be improved.

  In the said invention, it is desirable to provide an outer peripheral groove over the perimeter part of the said digging recessed part. With this configuration, when a microplate having a skirt portion provided on the periphery of a flat plate is fixed to the microplate holding portion, the microplate can be more stably inserted by inserting the skirt portion into the outer peripheral groove. It can be fixed to the microplate holding part.

In the above invention, it is desirable that the microplate holding portions formed on the rotor side wall are arranged at equal intervals in plan view around the rotation axis of the rotor.
With this configuration, the balance of the center of gravity of the rotor can be maintained even when the rotor is centrifugally accelerated.

Another invention is a centrifuge having a motor, a rotor connected to the motor and rotated, a chamber for housing the rotor, and a centrifuge lid for sealing the chamber. The centrifuge is characterized in that a microplate holding part formed of an assembly of a plurality of through holes into which each tube of the microtube assembly of the microplate can be inserted.
By comprising in this way, the outstanding centrifuge which can eliminate the difference of sample temperature and preset temperature small and light weight can be provided.

  According to the present invention, the centrifuge rotor can be reduced in size and weight, and the acceleration / deceleration time during centrifugal rotation can be shortened. Furthermore, the accuracy of experimental research can be improved by shortening the cooling time of the sample temperature in the microtube assembly and eliminating the difference from the set temperature.

It is a top view of the rotor of the centrifuge of this invention, and is a top view showing the state which hold | maintained one microplate to the rotor. It is BB sectional drawing of FIG. It is a side view of the rotor of the centrifuge of FIG. It is a PCR microplate (96 well), a in FIG. 4 represents a plan view, and b represents a side view. It is the figure which represents the dug recessed part which concerns on the centrifuge of this invention, and expanded the AA arrow line view of FIG. It is sectional drawing of the centrifuge incorporating the rotor of the centrifuge which concerns on this invention. It is a top view showing the state which provided three microplate holding | maintenance parts in the inner peripheral part of the rotor side wall of the centrifuge which concerns on this invention. It is a top view of the box-type centrifuge which applied the rotor of the centrifuge which concerns on this invention. It is EE sectional drawing of FIG. It is an 8 tube of a microplate, a is the top view, b is the FF sectional view, and c shows the side view. The modification of the digging recessed part which concerns on this invention is shown, a represents the partial enlarged plan view, b represents the expanded sectional view. It is a PCR microplate (96 wells) in which a skirt portion is formed on the outer periphery of a flat plate, a is a plan view thereof, b is a HH cross-sectional view thereof, and c is a side view thereof. It is another Example of the dug recessed part which concerns on the centrifuge of this invention, a is mounted | worn with the microplate holding part formed in the dug recessed part with the PCR microplate by which the skirt part was formed in the outer periphery of a plane plate The top view showing the state which carried out, b represents the DD sectional drawing. It is a figure showing the use method concerning the rotor of the present invention, and showing dehydration operation by attaching a microplate in the direction opposite to usual. These are sectional drawings showing the state which mounted | wore with the box-type sample holder on the inner surface of the rotor of the conventional centrifuge.

Mode for carrying out the invention

Hereinafter, a first embodiment of the present invention will be described with reference to the drawings.
First, an outline of a centrifuge incorporating a centrifuge rotor will be described with reference to FIG.
The centrifuge C includes a motor 17, a rotor 1 that is rotated by the motor 17 via a rotating shaft 18, a chamber 12 that houses the rotor 1, and a centrifuge lid 13 that seals the chamber 12. Prepare. The rotor 1 is fixed to the rotating shaft 18 by the cap screw 7. Further, the screw 5 is fixed to the bottom portion 2 of the rotor by four cap screws 6, and when the rotor lid 3 is used, the screw 5 is threaded so as to match the knob 19 for fixing the rotor lid 3. Is given. When fixing the lid 3 of the rotor, the knob 19 is rotated clockwise. The rotor lid 3 is installed to prevent heat generation due to rotational friction of the rotor and to serve as a protective wall when the microplate is damaged.

  When the rotating shaft 18 is rotated by the motor 17, the bottom 2 of the rotor 1 fixed to the rotating shaft rotates, and the rotor side wall 4 extending upward from the bottom of the rotor has a predetermined rotational speed (3000) in the chamber 12. To 15000 rpm).

  On the other hand, the temperature in the chamber rises due to the rotational friction of the rotor, but a cooling pipe 20 is wound around the outer periphery of the chamber 12 in order to suppress the temperature rise due to the rotational friction. It is carried out. A temperature sensor 21 for controlling the temperature in the chamber 12 is installed in the chamber 12 to control the temperature in the chamber and the sample temperature in the microtube assembly held on the rotor side wall. At the same time, the cooling pipe 20 cools the sample in the microtube assembly.

Next, the rotor of the centrifuge according to the embodiment of the present invention will be described.
FIG. 1 is a plan view of a rotor (hereinafter simply referred to as “rotor”) of the centrifuge of the present invention, and is a plan view showing a state in which one microplate is held by the rotor. FIG. It is -B sectional drawing. In the present embodiment, the rotor has an inverted truncated cone shape. The rotor 1 includes a bottom portion 2 fixed to a rotating shaft 18 of the rotor by a cap screw 7, and a rotor side wall 4 extending upward from the bottom portion at an inclined angle that is widened with respect to the rotating shaft 18. The inclination angle (θ) of the rotor side wall with respect to the rotation axis is manufactured in the range of 20 to 80 degrees. In the present embodiment, the rotor has an inverted truncated cone shape having an inclination angle of 60 degrees. The thickness of the rotor side wall is usually 5 mm to 30 mm, but is not particularly limited.
For the rotor, a hard and lightweight material selected from a reinforced resin, preferably a carbon fiber reinforced resin or a metal such as an aluminum alloy and / or a titanium alloy is used in order to shorten acceleration / deceleration time during centrifugal rotation. preferable.

Next, the PCR microplate 8 applied to this embodiment will be described with reference to FIG. This PCR microplate 8 has 96 wells (12 horizontal x 8 vertical), and a thin flat plate 8a (thickness 0.5 mm to 2.5 mm) is formed with an opening 8c for sample injection. Yes. The tubes 8b of the microtube assembly for injecting the sample protrude from the opening, and these tubes have a closed bottom. The length h of the protruding portion is prepared in various dimensions depending on the purpose of use. In this embodiment, the length h is 20 mm. The material of the PCR microplate is made of polystyrene, polyethylene or glass.
The wells of the PCR microplate used in this embodiment are 384, 1,536 well plates other than 96 wells, but are not limited thereto.

  Next, the microplate holding part R formed on the rotor side wall will be described. The rotor side wall 4 has a microplate holding portion R in which 96 through holes 11 corresponding to the number of wells of the PCR microplate are opened. These through holes are preferably perpendicular to the inner wall surface of the rotor. Has been opened. This is because when the microtube 8b is inserted into the through hole, no stress is applied between the flat plate 8a of the microplate and the microtube 8b due to the centrifugal acceleration force.

The vertical and horizontal pitch P2 between the through holes is designed to be equal to the vertical and horizontal pitch P1 of the opening of the PCR microplate. On the other hand, the inner diameter of the through hole 11 is slightly larger than the outer diameter of the largest portion of the tube 8b. That is, it is sufficient that the largest part of the microtube 8b can be inserted into the through hole 11 without resistance. Preferably, the inner diameter of the through hole is set to the outer diameter of the largest part of the tube 8b. On the other hand, it is better to be larger by 0.1 mm to 0.5 mm. If it is less than 0.1 mm, it cannot respond to the tube outer diameter currently marketed. On the other hand, if it exceeds 0.5 mm, the microplate may be deformed or broken by the centrifugal acceleration force, which is not preferable.
As described above, each tube 8 b of the microtube assembly can be smoothly inserted into the through hole 11.

  In the present embodiment, the number of through-holes 11 formed in the microplate holding part R is 96. The number of through-holes 11 is the number of wells of the PCR microplate and the vertical and horizontal pitch between the through-holes. It is determined by P2 and the vertical and horizontal pitches P1 of the openings of the PCR microplate. For example, in the case of 96 wells, if the vertical and horizontal pitch P2 between the through holes matches the vertical and horizontal pitch P1 of the opening of the PCR microplate, a PCR microplate having a smaller number of wells can be used as the microplate holder R. Needless to say, it can be retained.

Further, as is apparent from FIG. 2, the length h of the protruding portion of each tube 8b of the microtube assembly is shorter than the thickness of the rotor side wall. However, the reverse may be possible. In short, each tube of the microtube assembly may be inserted into the through hole to such an extent that the microplate is not detached from the microplate holding portion R by the centrifugal acceleration force.
As described above, the number of through-holes formed in the microplate holding part R, the pitch between the through-holes, and the diameter of the through-holes are appropriately determined according to the PCR microplate used.

In the present embodiment, the microplate holding portion R is provided in the dug recess 10 having a flat bottom surface 22 formed on the inner periphery of the rotor side wall 4. Thus, if the recessed part 10 is provided, a through hole is formed in the flat bottom surface 22 of the recessed part, and each tube 8b of the microtube assembly is inserted into this through hole, the flat plate 8a of the PCR microplate is dug. Close contact with the flat bottom 22 of the recess.
In particular, the rotor outer wall is not a planar shape, and is suitable for a rotor having a curved surface like the rotor of the present embodiment. Thus, by providing the dug recess 10, deformation of the microplate during centrifugal acceleration can be prevented.

Next, the operation of the centrifuge will be described.
As shown in FIG. 1, the microplate holding portions R formed on the rotor side wall are arranged symmetrically in plan view with the rotation shaft 18 as the center. Therefore, the center of gravity balance of the rotor can be maintained when centrifugal acceleration is performed. A sample to be separated is injected into each tube of the tube assembly of the PCR microplate. Samples are injected into two PCR microplates. The two microplates are mounted on the microplate holding portion R that is arranged symmetrically. When mounting, if each tube 8b of the tube assembly is inserted into the through-hole 11 formed in the bottom of the dug recess 10, the flat plate 8a of the microplate becomes a flat bottom surface 22 of the dug recess 10. Be in close contact with.

  Next, when the centrifugal acceleration is accelerated at, for example, 9000 rpm, the temperature in the chamber 12 increases due to the centrifugal acceleration, and the sample temperature also increases. However, a cooling pipe 20 is installed on the outer periphery of the chamber 12, and the temperature in the chin bar heated by centrifugal acceleration is controlled to a predetermined temperature by the temperature sensor 21. Therefore, the sample temperature injected into each tube of the microtube assembly is also controlled.

  Note that the sample in each tube of the microtube assembly is directly exposed in the chamber 12 at the tip of the microtube 8b into which the sample has been injected, so the difference between the temperature in the chamber and the sample temperature in the tube is extremely high. Small and can accurately control the actual sample temperature.

  After centrifugation, the microplate is firmly fixed to the microplate holder R, and the microplate does not move upward due to centrifugal acceleration. Further, since the flat plate 8a of the microplate is in intimate contact with the bottom surface of the dug recess 10, the microplate is not deformed or damaged, and the sample does not leak from the tube.

  In the present embodiment, two microplates into which a sample has been injected are prepared. However, in the case of one microplate, the other microplate holding part or the like is taken into account in consideration of the balance of the center of gravity of the rotating shaft during centrifugal acceleration. A weight dummy may be fixed.

  Further, the microplate holding portions R may be provided at symmetrical positions with respect to the rotation axis of the rotor as in this embodiment, or may be provided at equal intervals in three directions as shown in FIG. May be provided at equal intervals in four directions. Thus, even if rotational acceleration is applied to the rotor while the PCR microplate is held in the microplate holding region R, the center of gravity balance of the rotor can be maintained.

Next, as shown in FIG. 11, it is preferable to provide a notch S1 that communicates from the side wall 10a of the dug recess 10 formed on the inner periphery of the rotor side wall to the bottom 10b of the dug recess.
When this notch S1 is provided, a fingertip is inserted into this notch when the microplate that is in close contact with the microplate holding part is taken out by centrifugal acceleration, and the finger is placed on the back of the flat plate of the microplate and easily taken out. be able to. In addition, since no load is applied when the microplate is taken out, the sample in each tube after separation is not adversely affected.

Further, the microplate 30 as shown in FIG. 12 is obtained by hanging a skirt 30d on the side edge of the flat plate 30a of the microplate. When this microplate is used, as shown in FIG. It is desirable to provide an outer peripheral groove S2 into which the skirt portion 30d is inserted at the peripheral portion of the outer peripheral portion. By comprising in this way, it can fix stably to a microplate holding | maintenance part.
Furthermore, it is preferable to provide a notch S1 that communicates from the side wall 10a of the digging recess 10 formed on the inner periphery of the rotor side wall to the bottom 10b of the digging recess.

Next, FIG. 10 shows an 8-tube of a microplate. This microplate consists of an aggregate of eight tubes, and each microtube is connected by a flat plate 40a formed in the length direction of the tube 40b. When fixing this tube to the microplate holding part, it may be directly inserted and fixed to the microplate holding part formed on the inner peripheral surface, or may be inserted and fixed to the bottom of the digging recess. . In this case, the lower end of the flat plate 40a is in close contact with the rotor side wall or the bottom of the dug recess.

8 to 9 show another embodiment according to the present invention.
FIG. 8 is a plan view showing a state in which the microplate 8 (96 well) is mounted on the rotor 50 of the box-type centrifuge, and FIG. 9 is a cross-sectional view taken along line EE of FIG. The rotor 50 of the box-type centrifuge is connected to the bottom 52 fixed to the rotor rotating shaft 18, the flat rotor side wall 51 extending upward from the bottom 52, and the rotor side walls 51, 51. It consists of side plates 53 and 53 so as to withstand acceleration. The rotor side walls 51 and 51 are arranged with the rotational axis 18 as a center, and the center of gravity balance of the rotor can be maintained even when rotational acceleration is applied to the rotor.

Next, the microplate holding part R formed on the flat rotor side wall 51 will be described.
The flat rotor side wall 51 is formed with a microplate holding portion R formed of an assembly of 96 through holes 11 into which the microtubes 8b of the microtube assembly can be inserted on the inner peripheral portion thereof.

This through hole 11 is preferably formed perpendicular to the inner wall surface of the rotor. This is because when the microtube 8b is inserted into the through-hole, the microplate also applies no stress between the flat plate 8a and the microtube 8b due to centrifugal acceleration.
In this embodiment, it is not always necessary to provide the dug recess 10 as in the above embodiment, but when taking out the microplate from the microplate holding portion R after centrifugal acceleration, as in the above embodiment, When the thickness of the rotor side wall 51 is thicker than the microtube 8b, a notch S1 may be provided to easily take out the microplate.

FIG. 14 shows a different use for the rotor 1.
As shown in FIG. 14, a dehydration operation in the microtube 8 b can be performed by mounting the flat plate 8 a of the microplate 8 in the opposite direction to the normal so as to fit the recessed recess 10 of the rotor 1. Water droplets adhering to the inside of the microtube 8b by the centrifugal operation pass through the through hole 11 and are discharged into the chamber 12. The dehydration operation is mainly performed for the purpose of removing water droplets after washing the microplate 8, and is used at a low rotation (1000 rpm or less).

C Centrifugal separator 1 Rotor 2 Bottom 4 Rotor side wall 8 Microplate 8a Planar plate 8b Microtube R Microplate holding portion 10 Recessed recess 10a Side wall of recessed recess 10b Bottom of recessed recess S1 Notch S2 Outer peripheral groove 11 Through hole 12 Chamber 18 Rotating shaft 20 Cooling pipe 22 Flat bottom surface 21 Temperature sensor 30 Microplate 30a Flat plate 30d Skirt 40a Flat plate 40b Microtube 50 Rotor of box centrifuge 51 Flat rotor side wall 52 Bottom 53 Side plate 100 Box type sample Cage 110 Adapter 110a Movement prevention piece 400 Recessed insertion part 500 Movement stop

Claims (5)

  A microplate in which a flat plate and a microtube assembly into which a sample is injected and which is projected from the flat plate is integrally provided is held in a rotor of a centrifuge and rotated to be injected into the microtube assembly. In a centrifuge rotor for centrifuging a sample, the centrifuge rotor includes a bottom portion fixed to a rotating shaft of the rotor, and a rotor side wall extending upward from the bottom portion, and an inner peripheral portion of the rotor side wall. And a microplate holding portion formed of an assembly of a plurality of through holes into which the tube of the microtube assembly can be inserted.   The centrifuge rotor according to claim 1, wherein the microplate holding portion is formed in a dug recess having a flat bottom formed on an inner peripheral portion of the rotor side wall.   The rotor of the centrifuge according to claim 2, wherein a cutout portion that communicates from a side wall of the recessed portion to the bottom portion of the recessed portion is provided in the recessed portion.   The rotor of the centrifuge according to claim 2 or 3, wherein an outer peripheral groove is provided over the entire periphery of the peripheral edge of the dug recess.   A centrifuge having a motor, a rotor connected to the motor and rotated, a chamber for housing the rotor, and a centrifuge lid for sealing the chamber. A centrifuge in which a microplate holding part composed of an assembly of a plurality of through holes into which each tube of the tube assembly can be inserted is formed.

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