Centrifugal separation method
10420876 ยท 2019-09-24
Assignee
Inventors
Cpc classification
A61M1/3693
HUMAN NECESSITIES
B04B9/14
PERFORMING OPERATIONS; TRANSPORTING
B04B11/04
PERFORMING OPERATIONS; TRANSPORTING
B04B5/0407
PERFORMING OPERATIONS; TRANSPORTING
A61J1/05
HUMAN NECESSITIES
B04B1/02
PERFORMING OPERATIONS; TRANSPORTING
International classification
A61M1/36
HUMAN NECESSITIES
A61J1/05
HUMAN NECESSITIES
B04B1/02
PERFORMING OPERATIONS; TRANSPORTING
B01D17/02
PERFORMING OPERATIONS; TRANSPORTING
B04B5/04
PERFORMING OPERATIONS; TRANSPORTING
B04B9/14
PERFORMING OPERATIONS; TRANSPORTING
Abstract
A storage portion forming a storage space 10, includes an inclined inner wall portion 20 that is connected to a base portion so that the diameter of the inclined inner wall portion gradually decreases; a concave portion 22 is formed at a part of the inclined inner wall portion; and the concave portion 22 includes a concave portion side surface 22b that is connected to a concave portion bottom surface 22a. The concave portion 22 is formed at a position, where the concave portion crosses an interface S between the specimen centrifuged during rotation and air, in a radial direction with respect to the central axis; and the maximum width of the concave portion 22 in a circumferential direction around the central axis is included in a range of 2 mm to a length of 20% of the whole circumference.
Claims
1. A centrifugal separation method, using a centrifugal separation container, which centrifugally separating a first specific gravity component and a second specific gravity component, of which a specific gravity of the second gravity component is higher than a specific gravity of the first specific gravity component, contained in a specimen injected into a storage space by rotating a container about a central axis of the container as a rotation axis, the centrifugal separation method comprising: injecting a specimen into a storage portion that forms the storage space in the centrifugal separation container, wherein the storage portion includes an inclined inner wall portion of which a diameter gradually decreases from an upper end toward a lower end thereof and the lower end is connected to a base portion, and a concave portion formed at a part of the inclined inner wall portion, the concave portion includes a concave portion side surface that is connected to a concave portion bottom surface with a width of the concave portion gradually decreases toward the concave portion bottom surface from the inclined inner wall portion; rotating the centrifugal separation container about the central axis of the container as a rotation axis, wherein an interface between the specimen centrifuged during rotation and air, in a radial direction with respect to the central axis crosses where the concave portion is formed.
2. The centrifugal separation method according to claim 1, wherein a connecting portion between the inclined inner wall portion and the concave portion side surface has a curvature.
3. The centrifugal separation method according to claim 1, wherein a connecting portion between the concave portion side surface and the concave portion bottom surface has a curvature.
4. The centrifugal separation method according to claim 2, wherein a connecting portion between the concave portion side surface and the concave portion bottom surface has a curvature.
5. The centrifugal separation method according to claim 1, wherein a mean depth of the concave portion is 0.5 mm or more.
6. The centrifugal separation method according to claim 2, wherein a mean depth of the concave portion is 0.5 mm or more.
7. The centrifugal separation method according to claim 1, wherein the maximum depth of the concave portion is in the range of 0.5 mm to 2 mm.
8. The centrifugal separation method according to claim 1, wherein the maximum width of the concave portion is 10 mm or less.
9. The centrifugal separation method according to claim 1, wherein the concave portion has the shape of a fan.
10. The centrifugal separation method according to claim 1, wherein one concave portion is formed.
11. The centrifugal separation method according to claim 10, further comprising: a balancing portion that offsets a deviation of the center of gravity from the central axis caused by the formation of the concave portion to balance the container.
12. The centrifugal separation method according to claim 11, wherein the balancing portion is provided on the inclined inner wall portion at a position that is symmetrical to the position of the concave portion with respect to the central axis.
13. The centrifugal separation method according to claim 1, wherein two to four concave portions are formed.
14. The centrifugal separation method according to claim 13, wherein the concave portions are evenly disposed in the circumferential direction.
15. The centrifugal separation method according to claim 1, wherein the maximum length of the concave portion is in the range of 5 mm to 15 mm.
16. The centrifugal separation method according to claim 1, wherein the concave portion is formed from the uppermost portion of the inclined inner wall portion.
17. The centrifugal separation method according to claim 1, wherein the storage portion includes a trap portion that is connected to an upper end of the inclined inner wall portion and forms a trap space housing the second specific gravity component when the specimen is centrifugally separated and a layer structure body is formed on an outer peripheral side of the storage space.
18. The centrifugal separation method according to claim 1, wherein a thixotropic separating agent, which has a specific gravity between the specific gravity of the first specific gravity component and the specific gravity of the second specific gravity component, is provided in the storage space.
19. A centrifugal separation method comprising: the centrifugal separation method according to claim 1; and a step of holding the container by a container holder that rotates about a central axis of the container as a rotation axis.
Description
BRIEF DESCRIPTION OF THE DRAWINGS
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DESCRIPTION OF THE PREFERRED EMBODIMENTS
(12) An embodiment of the invention will be described below with reference to the drawings, but the invention is not limited to the embodiment. Meanwhile, for easy visual recognition, the scale and the like of each of components shown in the drawings are made to be appropriately different from the actual scale and the like.
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(14) The container 1 according to this embodiment includes the body member 2 and the lid member 3 of the container as shown in
(15) The container 1 has an internal structure substantially axisymmetric with respect to an axis (a central axis C of the container), which passes through the center of the base portion 21 and is perpendicular to the base portion 21, (in other words, a structure like a sort of rotating body having a center on the central axis C), and has a substantially columnar appearance as a whole. When centrifugal separation is performed, the lid member 3 is, for example, fixed to the fitting portion 24 while being fitted to the fitting portion 24 of the body member 2 and the container 1 is rotated about the central axis C as a rotation axis.
(16) When the body member 2 and the lid member 3 are fitted to each other, a storage space 10 into which a specimen is injected is formed as shown in
(17) The inclined inner wall portion 20 includes an inclined surface which is formed in a substantially conical shape and of which a diameter is reduced toward a lower end of the inclined surface connected to the base portion 21 from an upper end thereof connected to the trap bottom surface portion 23. A lower portion of the storage space 10 is formed by the inclined surface. Further, a part of the inclined inner wall portion 20 forms the concave portion 22. The concave portion 22 includes a concave portion side surface 22b that allows the width of the concave portion 22 to gradually decreases toward a concave portion bottom surface 22a from the inclined inner wall portion 20 and is connected to the concave portion bottom surface 22a. It is preferable that a connecting portion between the inclined inner wall portion 20 and the concave portion side surface 22b has a curvature to prevent the hemolysis of the specimen and a connecting portion between the concave portion bottom surface 22a and the concave portion side surface 22b has a curvature. Further details of the concave portion 22 will be described below. Further, the inclined inner wall portion 20 includes a protrusion 27 at a position symmetrical to the concave portion 22 with respect to the central axis C.
(18) The protrusion 27 is to adjust the position of the center of gravity of the container 1 that is changed due to the formation of the concave portion 22 of the inclined inner wall portion 20, and corresponds to a balancing portion of the invention. Only one concave portion 22 has been formed at an inclined inner wall portion 20 in this embodiment. However, since only one concave portion 22 is formed, the position of the center of gravity of the container 1 according to this embodiment may deviate from the central axis of the container in design. Since the rotation of the container 1 becomes unstable in this case, it is not preferable that only one concave portion 22 is formed. Accordingly, since the protrusion 27 is provided in this embodiment to offset a difference in the moment of inertia that is caused since a part (concave portion) of the inclined inner wall portion 20 becomes distant from the central axis C, the mass of a portion symmetrical to the position of the concave portion 22 with respect to the central axis C is increased. Further, the balancing portion only has to balance the container and is not limited to the protrusion-shaped structure. For example, a structure in which a high-density material is embedded in the inclined inner wall portion 20 at a position symmetrical to the position of the concave portion 22 with respect to the central axis C may be employed as the balancing portion. Furthermore, the balancing portion may not be provided on the inclined inner wall portion 20 and may be provided on the outer support wall portion 25. Meanwhile, the balancing portion is not essential in the invention, and does not need to be provided, for example, when the above-mentioned deviation of the center of gravity does not occur originally or when the deviation of the center of gravity is negligibly small in regard to centrifugal separation. Examples of a case in which the deviation of the center of gravity does not occur may include a case in which a plurality of concave portions are disposed so as to be balanced, and a case in which the thickness of a portion of the inclined inner wall portion 20 corresponding to a concave portion is adjusted to be small even if one concave portion is formed.
(19) The base portion 21 connected to the lower end of the inclined inner wall portion 20 includes a substantially horizontal flat surface that is connected to the lower end of the inclined surface of the inclined inner wall portion 20 so as to have a curvature. The flat surface forms the bottom surface of the storage space 10 in this embodiment, but the base portion 21 of the invention does not need to be flat and may be a rising curved surface. Since the container 1 is rotated about the central axis C, a specimen present near the central axis C tends to be hardly centrifugally separated. However, when the base portion 21 is formed of a rising curved surface, the centrifugal separation performance of the container can be further improved. The reason for this is as follows: when the base portion 21 is formed of a rising curved surface, a force in a direction away from the central axis C (which is a gravity component along the curved surface) is applied to a specimen present near the base portion 21 at the time of the injection of a specimen. As a result, since the specimen present near the base portion 21 is likely to be separated from the central axis C without staying near the central axis C during the rotation of the container 1, a centrifugal force is more efficiently applied to the specimen.
(20) The trap bottom surface portion 23 connected to the upper end of the inclined inner wall portion 20 includes a substantially horizontal flat surface that is connected to the upper end of the inclined surface of the inclined inner wall portion 20 so as to have a curvature. The flat surface forms the bottom surface of the trap space 10a. The trap side surface portion 26 includes a perpendicular surface that is connected to the flat surface of the trap bottom surface portion 23 so as to be perpendicular to the flat surface of the trap bottom surface portion 23. The perpendicular surface forms the side surface of the trap space 10a.
(21) The trap space 10a has an annular shape having a center on the central axis C, and the volume of the trap space 10a is designed according to the amount of a specimen to be injected. Further, a separating agent (or separation gel) is disposed, for example, in the trap space 10a in advance. The separating agent is appropriately selected from materials, which have an intermediate specific gravity between the specific gravities of the components, according to a low specific gravity component (first specific gravity component) and a high specific gravity component (second specific gravity component) that are contained in the specimen and are to be separated. Specifically, when blood plasma (low specific gravity component) and blood cells (high specific gravity component) contained in blood are to be separated, a material having an intermediate specific gravity between the specific gravity of the blood plasma and the specific gravity of the blood cell may be selected as the separating agent.
(22) The outer support wall portion 25 extends downward from the trap side surface portion 26 while surrounding the entire inclined inner wall portion 20, and a lower end of the outer support wall portion 25 extends downward further than the base portion 21. Accordingly, the body member 2 is stably supported.
(23) The opening portion 30 of the lid member 3 has, for example, a truncated conical shape and includes an inclined surface of which a diameter is reduced toward the opening 31. An upper portion of the storage space 10 is formed by the inclined surface. In this embodiment, the container 1 is rotated while the opening 31 is open. However, the opening may be adapted to be capable of being opened and closed as necessary. The trap upper surface portion 33 connected to the lower end of the opening portion 30 includes a substantially horizontal flat surface that is connected to the lower end of the inclined surface of the opening portion 30 so as to have a curvature. The flat surface forms the upper surface of the trap space 10a.
(24) Further, centrifugal separation is performed by, for example, a centrifugal separation device 50 shown in
(25) The concave portion 22 of the inclined inner wall portion 20 will be described.
(26) It is preferable that the shape of the concave portion 22 is a fan shape having a center on the central axis C (including a truncated fan shape of which a portion including a central portion is cut out) so that an obstacle occurring when the specimen is centrifuged or when the low specific gravity component moves downward along the inclined surface of the inclined inner wall portion 20 is reduced. A fan shape (circular sector) of the invention means a shape of which at least a part includes a side parallel to a radial direction in a top view, and each corner of the fan shape may be rounded. Further, it is preferable that a side, which mainly extends in the circumferential direction around the central axis C, has an arc shape in the circumferential direction. Meanwhile, the concave portion 22 includes the flat bottom surface 22a along the inclined surface of the inclined inner wall portion 20 in this embodiment, but the bottom of the concave portion 22 is not limited to a flat surface.
(27) The maximum width W1 (
(28) Further, the maximum length W2 (
(29) Processes of a centrifugal separation method using the centrifugal separation container 1 and the centrifugal separation device 50, which have been described above, will be described below.
(30) First, the container 1 in which the separating agent 4 is disposed in the trap space 10a in advance is prepared, and a specimen 5 is injected into the storage space 10 from the opening 31 of the container 1 (
(31) As described above, according to the container of this embodiment, a portion of the low specific gravity component, which is present above the concave portion, is more easily released from the layer structure body than portions of the low specific gravity component that are present in other regions. Accordingly, a portion of the low specific gravity component, which is present above the concave portion, is released first from the layer structure body, and portions of the low specific gravity component, which are present in other regions, are released from the layer structure body so as to be pulled to the portion of the low specific gravity component, which is present above the concave portion, due to the release of the portion of the low specific gravity component present above the concave portion. As a result, centrifugally separated components of the specimen can be more efficiently recovered. Further, the concave portion can be easily formed without other members simultaneously with the molding of the container. For this reason, it is possible to reduce manufacturing cost by simplifying processes for manufacturing the container.
(32) Furthermore, since centrifugal separation is performed using the centrifugal separation container of the invention in the centrifugal separation device and the centrifugal separation method of the invention, centrifugally separated components of the specimen can be more efficiently recovered.
OTHER EMBODIMENTS
(33) The body member or the concave portion of the container 1 of the invention is not limited to the structure of the body member or the concave portion of the above-mentioned embodiment.
(34) For example,
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(36) Furthermore,
(37) Further, the fan-shaped concave portion having a center on the central axis C has been described in the above-mentioned embodiment and
EXAMPLES
(38) Examples of the centrifugal separation method using the centrifugal separation container of the invention will be described below.
Example 1
(39) A centrifugal separation container shown in
(40) Diameter 1 of a trap space=22.5 mm
(41) Diameter 2 of the circumference including a balancing protrusion=14 mm
(42) Diameter 3 of the entire container=26 mm
(43) Height L1 of a body member=18.1 mm
(44) Depth L2 of a space formed by an inclined inner wall portion=9 mm
(45) Height L3 of the trap space=4.8 mm
(46) Depth D of a concave portion=0.8 mm
(47) Angle 1 between an inclined surface of the inclined inner wall portion and a central axis=48
(48) A circumferential angular range 2 occupied by the concave portion=47
(49) A distance R between a base portion and the concave portion along the inclined surface of the inclined inner wall portion=4.1 mm
(50) Further,
(51) 600 L of SCOLLECT (manufactured by Sekisui Medical Co., Ltd.) was dispensed to the centrifugal separation container as a serum separating agent, and 500 L of whole blood was dispensed. After that, a specimen was centrifuged for 90 seconds (for 110 seconds when 10 seconds at the time of acceleration and 10 seconds at the time of deceleration are included) at a rotational speed of 24000 rpm. Further, the centrifugal separation method was performed twice.
Example 2 and Comparative Example
(52) Centrifugal separation was performed using a container, which included two concave portions, as in Example 2. The concave portions were provided at positions that face each other with a central axis interposed therebetween. Other conditions were the same as those of Example 1. Furthermore, centrifugal separation was performed using a container, which did not include a concave portion and of which an inclined surface of an inclined inner wall portion was flat, as Comparative example. Other conditions were the same as those of Example 1.
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EXPLANATION OF REFERENCES
(55) 1: centrifugal separation container
(56) 2: body member
(57) 3: lid member
(58) 4: separating agent
(59) 5: specimen
(60) 5a: low specific gravity component (first specific gravity component)
(61) 5b: high specific gravity component (second specific gravity component)
(62) 10: storage space
(63) 10a: trap space
(64) 20: inner wall portion
(65) 21: base portion
(66) 22: concave portion
(67) 22a: concave portion bottom surface
(68) 22b: concave portion side surface
(69) 23: trap bottom surface portion
(70) 24: fitting portion
(71) 25: outer support wall portion
(72) 26: trap side surface portion
(73) 27: protrusion
(74) 30: opening portion
(75) 31: opening
(76) 33: surface portion
(77) 50: centrifugal separation device
(78) R: rotation axis
(79) S: interface
(80) W1: maximum width
(81) W2: maximum length