JP2002538480A - Method and apparatus for moving fluid by multiple centrifugations - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide at least one arrival room (6) from at least one departure room (4) via at least one movement conduit (9 and 10) whose movement is effected by centrifugal force. Method of moving fluid (2) to the air and an apparatus utilizing the same. A first centrifuge (C1), called primary, for moving a fluid (2) from its starting chamber (4) to an intermediate chamber (5) via a primary transfer conduit (9). -) That of the first centrifugation to move the fluid (2) from the intermediate chamber (5) to the arrival chamber (6) via the secondary transfer conduit (10); A method comprising performing at least one second centrifugation (C2), called secondary, wherein the centrifugation axis (8) differs from (7), and the intermediate chamber (5) has at least 2 Two arrival chambers (6), each arrival chamber (6) being connected to the intermediate chamber (5) by a secondary transfer conduit (10), and for the fluid present in said intermediate chamber (5) to be at least two. At least one time allowing to direct towards the arrivals room (6) An apparatus, comprising performing a secondary centrifugation.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD OF THE INVENTION

The present invention relates to a method and a device for moving a fluid, in particular a liquid, from a departure chamber to an arrival chamber via a movement conduit, the movement being effected by the action of centrifugal force.

[0002]

[Prior art]

The state-of-the-art is directed to the publication FR which deals with the movement of liquid in a device for collecting and returning a predetermined amount of liquid.
-A-2.678.379. The device is tubular and comprises a single-eye helical conduit having a centrifugal axis as the axis of symmetry near the periphery. When the device is positioned along the axis of rotation with the opening of the internal conduit in contact with the liquid, the liquid rises in the device, and in the opposite direction, the liquid emerges therefrom.

[0003]

[Problems to be solved by the invention]

This device can only collect and distribute the collected liquid. There is no possibility to direct the liquid inside. The versatility of this system is very limited, and further limited to maintaining centrifugation as the harvest remains in the device.

[0004] According to another embodiment, centrifugation can be more versatile, and thus can be incorporated into devices that allow one to carry out multiple biological reactions. Thus, there are a large number of documents that are fairly close to each other. For example, U.S. Pat.
SA-3,744,975, US-A-4,123,173 and US-A-
4,225,558 correspond to it. This axis therefore makes it possible to move the liquid contained in the device from the center of the device to its periphery under the action of centrifugal force.

However, it is not conceivable to carry out the movement in the opposite direction, ie according to the centripetal force. Therefore, the liquid can be oriented only in the centrifugal direction. Publications US-A-4,812,294, EP-A-0.297.394 and US
-A-4, 788, 154 describes an apparatus and method that allows movement of a liquid along a plurality of directions in response to centrifugal forces exerted by a plurality of centrifugal axes.

[0006] Nevertheless, none of these documents has an axis in the direction in which the centrifugal force is applied, on the one hand, first, the departure or intermediate chamber, then, the intermediate or arrival chamber, and, on the other hand, these chambers. The configuration of the conduits between them. Furthermore, the invention according to the application distributes the liquid in a ratio set according to the shape and the presence of at least two intermediate chambers for one departure chamber and / or at least two arrival chambers for one intermediate chamber. enable.

[0007]

[Means for Solving the Problems]

According to the invention, its use is even more versatile, allowing the movement in all chambers of the device and the distribution of liquid in a set ratio without the effects of a single centrifugation. The invention also relates to a method of using such a device. The invention therefore provides a method of transferring fluid from at least one departure chamber to at least one arrival chamber via at least one transfer conduit, wherein the transfer is effected by the action of centrifugal force: Performing a first centrifugation, referred to as primary, to transfer the fluid from its departure chamber to the intermediate chamber; and-transferring the fluid from the intermediate chamber to the destination chamber via a secondary transfer conduit. To move
Performing at least one second centrifugation, called secondary, which is different in axis from that of the first centrifuge, wherein the intermediate chamber is combined with at least two arrival chambers, each of which has a secondary movement Connected to the intermediate chamber by a conduit, wherein at least two fluids are present in said intermediate chamber.
Performing at least one secondary centrifugation that allows it to be directed toward one of the arrival chambers.

[0008] In the present invention or a fluid transfer device comprising a body with at least one departure chamber, at least one arrival chamber and at least one transfer conduit, from the departure chamber to the intermediate chamber via the primary transfer conduit. At least two of at least one secondary axis, different from the primary axis, for moving fluid from the intermediate chamber to the arrival chamber via a secondary transfer conduit, The invention also relates to a device characterized in that it also comprises two centrifuge shafts. In this apparatus, each centrifugal axis is a chamber in which a fluid is present, and a chamber in which liquid is sent by centrifugal separation along an associated centrifugal axis, wherein the chamber in which a fluid is present; It is characterized in that the virtual axis through the chamber located between the edges of the device is cut substantially perpendicularly.

[0009] According to a variant embodiment, the respective transfer conduit between the two chambers is located approximately along an imaginary axis through the two chambers on both sides of the conduit. According to another variant embodiment, each transfer conduit between the two chambers is associated with one centrifuge shaft.

According to yet another embodiment, each transfer conduit is straight and passes through the center of gravity of two chambers located on both sides of said transfer conduit.

According to a similar variant, each transfer conduit is provided with a blocking means, such as a ball valve, for blocking the passage of the displaced or displaced fluid.
According to another variant embodiment, the intermediate room is connected to at least two adjacent arrival rooms, each arrival room being connected to the intermediate room by a secondary transfer conduit, these arrival rooms being located between the adjacent arrival rooms. Is combined with a single centrifuge shaft that allows for the distribution of

According to a different variant embodiment, the intermediate chamber is directed by centrifugation in the area of the intersection area with the secondary transfer conduit, which corresponds to the adjacent arrival chamber, of the starting fluid in the area of said conduit. And a shape that allows equal distribution between said adjacent arrival rooms.

[0013] Such a device follows all of the simple or complex steps of an analysis using one or more different reagents depending on the chemical, physical or biological properties of the one or more analytes studied. It can be used to analyze one or more different liquid specimens that seek to identify one or more analytes. The technical principles defined below are not limited to a particular analyte, the only requirement being that the analyte be distributed in the sample to be analyzed in suspension or solution. In particular, the analytical methods performed can be performed in a homogeneous, heterogeneous or mixed form.

[0014] A non-limiting, specific aspect of such a device relates to the biological analysis of one or more ligands, the detection and / or quantification of which requires the use of one or more antiligands. It is. Ligand shall mean any biological species such as, for example, antigens, antigen fragments, peptides, antibodies, antibody fragments, haptens, nucleic acids, nucleic acid fragments, hormones, vitamins and the like. One application of analytical techniques involves immunoassays, in any form, by direct analysis or by competition. Another application is
It relates to the detection and / or quantification of nucleic acids, including the entire work required for detection and / or quantification from any collection containing target nucleic acids. Among these different tasks are the steps of digestion, mobilization, concentration, enzymatic amplification of nucleic acids, for example detection steps including hybridization steps using DNA chips or labeled probes. Publication WO-A-97 / 02357 discloses the different steps required in the case of nucleic acid analysis.

[0015]

BEST MODE FOR CARRYING OUT THE INVENTION

The accompanying figures are given by way of example and are not meant to be limiting in any way. They allow a better understanding of the invention.

The present invention relates to a novel method for moving the liquid 2 in the moving device 1 constituted by the main body 3. As is evident from these figures, the device 1 has a substantially parallelepiped shape, even if only the upper surface of the device or card 1 is visible. For implementation, the card is compatible with the processing solution, eg, GOODDFELL
Obtained by processing technical plastics, such as OW number R540E impact polystyrene. In an industrial embodiment, the card can be obtained by precision molding, but all other manufacturing methods, in particular the patent application WO-A-97 / 02357.
Those used in semiconductor technology, such as those described in, can be used in the manufacture of such cards.

1 to 3 show a first embodiment. The purpose is to enable the movement of the specimen 2 contained in the departure room 4 to the four arrival rooms indicated by reference numeral 6. The apparatus 1 therefore comprises a first departure room 4 located on the left of these figures. It is possible to pass through an intermediate chamber 5 having a shape of substantially "green beans",
This intermediate room 5 is centrally located from the viewpoint of movement between the departure room 4 and the four arrival rooms 6 in these figures. However, chamber 6 is the position occupied by intermediate chamber 5, which is not located to the right of the whole figure. The physical location of the room will be detailed later in connection with the different steps of the movement method. Of course, these conduits are present to allow movement from chamber to chamber. First the primary transfer conduit connecting the departure room 4 to the intermediate room, and finally the intermediate room 5 to the arrival room 6
There is a secondary transfer conduit 10 that connects to

In the embodiment of FIGS. 1-3, each arrival room 6 has one secondary transfer conduit 1
0, and the entirety of the conduit 10 and the chamber 6 is independent for each arrival chamber 6. The overall arrangement of the rooms is such that the departure room 4 is located to the left, the intermediate room 5 is arranged to the right and the arrival room 6 is located between these two rooms 4 and 5. Therefore, movement is not possible without achieving multiple centrifugation. This is what these figures 1
To 3 are shown.

For example, in FIG. 1, the liquid specimen 2 exists only in the region of the departure room 4. In this position, the liquid specimen 2 has just been placed in this chamber 4 or is actually in its initial position, since an independent travel channel carried it to this chamber 4. According to FIG. 2, the primary centrifugation is carried out along C1 around the primary centrifuge axis 7. In this case, the liquid 2 is carried into the intermediate chamber 5 via the primary movement chamber 9 along the arrow F1. As clearly shown in this figure, the liquid 2 is located as far as possible from the shaft 7 in the region of this intermediate chamber 5.

According to FIG. 3, a second centrifugation is carried out along C 2 in the region of the secondary centrifugal axis 8. When the entire device 1 is rotated along C2, as can be easily understood,
At this time, the liquid 2 is carried into the arrival room 6 through the secondary moving conduit 10. This movement is performed along the arrow F2 in FIG. Here too, the liquid specimen 2 distributed in the arrival chamber 6 is located as far as possible from the centrifuge shaft 8.

The shape of the intermediate chamber is quite special in the area of the installation point of the secondary transfer conduit 10 so that the liquid specimen 2 in the area of the intermediate chamber 5 does not return towards the starting chamber 4. That is, as indicated above, the intermediate chamber 5 has a substantially "green bean" shape such that there are two-lobed leaves on one of the sides of the chamber 5. These two lobes surround the intersection of the intermediate chamber 5 with the conduit 9. Each application is combined with two conduits 10, where the shape of the chamber 5 prevents, or at least minimizes, the passage of liquid into the conduits 9 and the second along C2 to the arrival chamber 6 Enables the orientation and movement of liquid during centrifugation.
Of course, a valve, not shown, can be located above conduit 9 to prevent liquid reflux if necessary. In order to minimize this liquid reflux in conduit 9, the volume of liquid contained in the lower lobe of chamber 5 is moved into two chambers 6 located below conduit 9 in FIGS. Must exceed the amount of liquid you want. The volume of the lobes of the chamber 5 is, on the one hand, perpendicular to the conduit and is clearly defined by the ray positioned at the end of this conduit 9 and, on the other hand, by the intersection between the same chambers 5 of the conduit 10 bed. The same rules apply to the upper leaves. Preferably, in order to avoid fluid communication between the two chambers 6, the volume of the lobes should be less than the total volume of the two chambers 6 and the two conduits 10 associated with said chambers. In one embodiment, the two lobes have the same volume. In another embodiment, the two lobes have different volumes. In one embodiment, the volumes of the arrival rooms are the same. The shape and dimensions of the conduit 10 are selected by those skilled in the art to achieve a homogeneous distribution of the liquid in the different arrival chambers. In another embodiment, the volumes of the arrival rooms are different.

The total volume of liquid displaceable by this device can vary from 0.5 to 5000 microliters, preferably from 5 to 1000 microliters.
The volume of the starting chamber can vary in the same ratio, or can be much larger than the total amount moved.

By way of example, in the embodiment of FIGS. 1 to 3, the chamber 4 has a volume of 0.1 microliters for moving 100 microliters of liquid towards an intermediate chamber 5 approximately equal to that of the chamber 4.
And a volume comprised between 0.5 ml. During the second centrifugation along axis C2, 25 microliters are distributed to each arrival chamber 6 with an accuracy of less than 5%.

Of course, these shapes and the number of conduits and chambers are not limiting at all,
Other shapes of the card 1 as well as different numbers of chambers and conduits are possible. The thing is,
FIG. 4 also shows that the card 11 is substantially square when viewed from below. At this time, there is a departure room 14 at the center of the main body 13, which is connected to four intermediate rooms 15 via four primary transfer conduits 19. In order to facilitate the operation of the centrifuge and the distribution of the liquid according to the user's choice, the four intermediate chambers are arranged symmetrically with respect to each other. Next to the centrifugal shaft 18,
There are also two other centrifuge shafts 12. All of these axes have specific positions. Thus, by taking the center of the departure chamber 14 and drawing a half-line from this center toward the center of gravity of each of the intermediate chambers 15, each centrifugal shaft 17, 18 or 12 will It is arranged on this half line at a position located between the edges.

Similarly, there is also an arrival chamber 16 connected to the intermediate chamber 15 by a secondary transfer conduit 20. As can be seen, these secondary transfer conduits 20 have an intersection with the intermediate chamber 15, through which a straight line passing through the center of gravity of the intermediate chamber 15 can be passed. This straight line is, in fact, substantially perpendicular to the half-line from the center of gravity of the starting chamber 14 to the center of gravity of the intermediate chamber 15. As can be easily understood here, it is possible to select from the liquid specimen located in the center, ie in the region of the starting chamber 14, an intermediate chamber, not shown in FIG. it can. For example, when the apparatus 11 is rotated along the centrifugal axis 17, the liquid 2 is moved toward the intermediate chamber 15 located on the left of FIG. When the liquid 2 pixels are reached, the liquid 2 can be centrifuged along the centrifugal separation axis 18 in order to move the liquid 2 from the intermediate chamber 15 to the arrival chamber 16 arranged at the upper position. . Conversely, if it is desired to move the liquid towards the lower arrival chamber 16, it will be necessary to create a centrifugation on the centrifuge shaft 12 arranged in the upper position. Similarly, it is also conceivable that the liquid 2 needs to be moved again towards the departure chamber 14 or even further towards the intermediate chamber 15 located to the right. In this case, it is necessary to centrifuge along the axis 12 located on the left side of the figure. Thus, it is readily appreciated that the versatility of such a system and the number of possibilities for movement within the same card is very large.

In FIG. 5, a third, last embodiment of the device 21 is shown. This is almost identical to that of FIG. 4, but it can be seen that once the liquid arrives in one area of the arrival chamber 26, there is another possibility. Thus, each arrival chamber 26 is associated with two end transfer conduits 32 connecting it to the two end chambers 21.
In order to obtain the liquid 2 present in the end chamber 31 shown in FIG.
The specimen is placed in a starting chamber 24 located at the center of 3 and then centrifuged along 27 to move the specimen 2 towards an intermediate chamber 25 located on the left of the figure, 28 to move the specimen from room 25 to arrival room 26
It is necessary to continue the centrifugation along 28 in order to allow movement of the liquid 2 in this end chamber 31.

Of course, it is conceivable to use liquid stopping means in the area of each conduit to better control the movement of the liquid 2 that may be introduced. For example, the capillary action of these conduits can be used, but valves can be provided in those areas to open and close the conduits. Such a valve is described in detail in a patent application filed by the applicant on Sep. 8, 1998 under application number FR 98/11383, entitled "Devices for enabling reactions, transfer systems between devices and methods of using such systems". Has been described.

Similarly, although not shown in FIGS. 1 to 4, in order to ensure good operation, the device according to the invention is provided with a fluid inlet and / or outlet. For centrifugation to be effective, it need not be very fast. 2 and 10g (
A centrifugation comprised between the sign of gravity acceleration) or preferably between 3 and 5 g is quite sufficient for non-viscous biological liquids. For viscous liquids, liquids comprised between 10 and 200 g can be applied.

In order to make the centrifugation as efficient as possible, it is possible to have as many centrifugation axes as there are transfer conduits between two adjacent chambers between which one wants to transfer liquid. For example, FIGS. 4 and 5 show only four axes 27, 28, and 22. It is also possible to envisage it in the corner area of the respective device 11 or 21 in order to facilitate movement towards the arrival chamber 16 or 26 and further towards the end chamber 31. Furthermore, the conduits are preferably straight, also to increase the effectiveness of these centrifugations.

[Brief description of the drawings]

FIG. 1 shows an elevational view of a first embodiment of the present invention before an initial centrifugation.

FIG. 2 shows an elevation view of the first embodiment of the invention after the first centrifugation and before the second centrifugation.

FIG. 3 shows an elevation of the first embodiment of the present invention after a second centrifugation.

FIG. 4 shows an elevation view of a second embodiment of the present invention.

FIG. 5 shows an elevation view of a third embodiment of the present invention.

[Explanation of symbols]

 1. 1. Mobile device according to first embodiment Fluid or liquid Main body 4. Departure room 5. Intermediate room 6. Arrival room 7. Primary centrifuge shaft 8. Secondary centrifuge shaft 9. Primary moving conduit 10. Secondary moving conduit 11. 11. Mobile device according to second embodiment 12. Other centrifugation shafts Body 14. Departure room 15. Intermediate room 16. Arrival room 17. Primary centrifuge shaft 18. Secondary centrifuge shaft 19. Primary moving conduit 20. Secondary moving conduit 21. Moving device 22. Other centrifuge shafts 23. Main body 24. Departure room 25. Intermediate room 26. Arrival room 27. Primary centrifuge shaft 28. Secondary centrifuge shaft 29. Primary transfer conduit 30. Secondary moving conduit 31. Terminal chamber 32. Terminal transfer conduit C1. Rotational motion of device 1 for driving the first centrifuge C2. Rotational motion of the device 1 for driving the second centrifuge F1. Movement of liquid 2 by action of first centrifugation F2. Movement of liquid 2 by action of second centrifugation

[Procedural Amendment] Submission of translation of Article 34 Amendment

[Submission Date] March 13, 2001 (2001.3.13)

[Procedure amendment 1]

[Document name to be amended] Statement

[Correction target item name] Claims

[Correction method] Change

[Contents of correction]

[Claims]

──────────────────────────────────────────────────続 き Continuation of front page (81) Designated country EP (AT, BE, CH, CY, DE, DK, ES, FI, FR, GB, GR, IE, IT, LU, MC, NL, PT, SE ), OA (BF, BJ, CF, CG, CI, CM, GA, GN, GW, ML, MR, NE, SN, TD, TG), AP (GH, GM, KE, LS, MW, SD, SL, SZ, TZ, UG, ZW), EA (AM, AZ, BY, KG, KZ, MD, RU, TJ, TM), AE, AL, AM, AT, AU, AZ, BA, BB, BG, BR, BY, CA, CH, CN, CU, CZ, DE, DK, EE, ES, FI, GB, GD, GE, GH, GM, HR, HU, ID, IL, IN , IS, JP, KE, KG, KP, KR, KZ, LC, LK, LR, LS, LT, LU, LV, MD, MG, MK, MN, MW, MX, NO, NZ, PL, PT, RO, RU, SD, SE, SG, SI, SK, SL, TJ, TM, TR, TT, UA, UG, US, UZ, VN, YU, ZA, ZW

Claims (8)

[Claims] At least one moving conduit (1), whose movement is effected by centrifugal force.
Transfer of fluid (2) from at least one departure room (4, 14 or 24) to at least one arrival room (6, 16 or 26) via 9 and 10, 19 and 20 or 29 and 30) In the method, for moving the fluid (2) from its departure chamber (4, 14 or 24) to the intermediate chamber (5, 15 or 25) via a primary transfer conduit (9, 19 or 29) Performing a first centrifugation (C1), called the primary; from the intermediate chamber (5, 15 or 25) via the secondary transfer conduit (10, 20 or 30) to the arrival chamber (6, 16). Or 26) to move the fluid (2) to at least one second centrifuge (C2), which is different from the first centrifuge (7) and the centrifugal axis (8) is different from the second centrifuge (C2). ) Including the steps of: , 15 or 25) are combined into at least two arrival chambers (6, 16 or 26), each arrival chamber (6, 16 or 26) being connected by an intermediate transfer conduit (10, 20 or 30) to an intermediate chamber (5 or 25). , 15 or 25) at least once to allow the fluid present in said intermediate chamber (5, 15 or 25) to be directed towards at least two arrival chambers (6, 16 or 26) Performing a secondary centrifugation (C2) of the method. 2. At least one departure room (4, 14, or 24), at least one arrival room (6, 16 or 26) and at least one transfer conduit (9, 1).
9 or 29 or 10, 20, 20 or 30) (3, 13 or 2)
In the transfer device (1, 11 or 21) for the fluid (2) containing 3), from the starting chamber (4, 14, or 24) via the primary transfer conduit (9, 19 or 29) to the intermediate chamber (5). , 15 or 25) for moving the fluid (2) toward the primary axis (
7, 17 or 27) and a fluid (6, 16 or 26) from the intermediate chamber (5, 15 or 25) to the arrival chamber (6, 16 or 26) via a secondary transfer conduit (10, 20 or 30).
2) for moving at least two centrifugal axes (7 and 8, 17) of at least one secondary axis (8, 18 or 28) different from the primary axis (7, 17 or 27);
And 18 or 27 and 28), and the respective centrifugal axis (7 and 8, 17 and 18, or 27 and 28) provides the centrifugal action along the chamber in which the fluid is present and the associated centrifuge axis. A chamber into which liquid is to be received and into which the fluid is present, and the device (1, 11 or 2)
An apparatus characterized by cutting a virtual axis through a chamber positioned between the edges of 1) substantially perpendicularly. 3. The apparatus according to claim 2, wherein each of the moving conduits (9 or 10, 19 or 20 or 29 or 30) between the two chambers is provided with a corresponding one of the conduits (9 or 10, 19 or 20). , Or 29 or 30), positioned substantially along an imaginary axis passing through the two chambers. 4. The device according to claim 2, wherein the two chambers (4, 14 or 24 or 6, 16 or 26, and 5, 15).
Or 25) each transfer conduit (9 or 10, 19 or 20 or 29 or 30) is combined with one centrifugal shaft (7 and 8, 17 and 18 or 27 and 28) An apparatus characterized by the above. 5. The device according to claim 2, wherein each of the moving conduits (9 or 10, 19 or 20, or 29 or 30) is straight and the moving conduit (9 or 30) is straight. 10, 19 or 20, or 29 or 30), two chambers (4, 14 or 24 or 6,
16 or 26, and 5, 15 or 25). 6. An apparatus according to claim 2, wherein each of the moving conduits comprises a blocking means, such as a ball valve, for blocking the passage of the displaced or displaced fluid. An apparatus characterized in that: 7. The device according to claim 2, wherein the intermediate chambers (5, 15 or 25) are adjacent to at least two arrival chambers (6.1).
6 or 26) and their respective arrival chambers (6, 16 or 26) are connected by secondary transfer conduits (10 20 or 30) to the intermediate chambers (5, 15 or 25) and these arrival chambers (6 , 16 or 26) is combined with a single centrifugal shaft (8, 18 or 28) allowing a distribution between adjacent arrival chambers (6, 16 or 26). 8. The apparatus according to claim 7, wherein the intermediate chamber (5, 15 or 25) is adjacent to the arrival chamber (6, 16 or 26).
The direction of the starting fluid (2) in the area of said conduit (10 20 or 30) by centrifugation (C1 or C2) in the area of the intersection area with the secondary transfer conduit (10 20 or 30), corresponding to And the adjacent arrival room (6, 16 or 26)
The device characterized in that it has a shape that allows an equal distribution between the two.