EP1239962B1 - Dispositif de microanalyse - Google Patents
Dispositif de microanalyse Download PDFInfo
- Publication number
- EP1239962B1 EP1239962B1 EP00992452A EP00992452A EP1239962B1 EP 1239962 B1 EP1239962 B1 EP 1239962B1 EP 00992452 A EP00992452 A EP 00992452A EP 00992452 A EP00992452 A EP 00992452A EP 1239962 B1 EP1239962 B1 EP 1239962B1
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- European Patent Office
- Prior art keywords
- chamber
- fluid
- arm
- microstructure
- disc
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- the present invention relates to microanalysis devices and methods for moving fluids in such devices.
- micro-analysis systems that are based on microchannels formed in a rotatable, usually plastic, disc, often called a "centrifugal rotor" or "lab on a chip".
- Such discs can be used to perform analysis and separation on small quantities of fluids.
- the discs should be not restricted to use with just one type of reagent or fluid but should be able to work with a variety of fluids.
- the disc permits the user to dispense accurate volumes of any desired combination of fluids or samples without modifying the disc.
- any air bubbles present between two samples of fluids in the microchannels can act as separation barriers or can block the microchannel and thereby can prevent a fluid from entering a microchannel that it is supposed to enter.
- US patent no. 5 591 643 teaches the use of a centrifugal rotor which has microchannels that have cross sectional areas which are sufficiently large that unwanted air can be vented out of the microchannel at the same time as the fluid enters the microchannel.
- An object of the present invention is to provide a structure for a centrifugal rotor and a method for using such a centrifugal rotor, which structure and which method permits the reliable transport of fluids in the centrifugal rotor.
- a further object of the present invention is to provide a structure for a centrifugal rotor and a method for using such a centrifugal rotor, which structure and which method permits the accurate metering of fluids in the centrifugal rotor.
- the present invention achieves the objects of the invention by means of a structure having the features of claim 1.
- a method for using such a structure to achieve the objects of the invention has the features of claim 5.
- microfluidic disc is of a one- or two-piece moulded construction and is formed of an optionally transparent plastic or polymeric material by means of separate mouldings which are assembled together (e.g. by heating) to provide a closed structure with openings at defined positions to allow loading of the device with fluids and removal of fluid samples.
- Suitable plastic of polymeric materials may be selected to have hydrophobic properties.
- Preferred plastics or polymeric materials are selected from polystyrene and polycarbonate.
- the surface of the microchannels may be additionally selectively modified by chemical or physical means to alter the surface properties so as to produce localised regions of hydrophobicity or hydrophilicity within the microchannels to confer a desired property.
- Preferred plastics are selected from polymers with a charged surface, suitably chemically or ion-plasma treated polystyrene, polycarbonate or other rigid transparent polymers.
- the microchannels may be formed by micro-machining methods in which the micro-channels are micro-machined into the surface of the disc, and a cover plate, for example, a plastic film is adhered to the surface so as to enclose the channels.
- the microfluidic disc (D) has a thickness which is much less than its diameter and is intended to be rotated around a central hole so that centrifugal force causes fluid arranged in the microchannels in the disc to flow towards the outer periphery of the disc.
- the microchannels start from a common, annular inner application channel (1) and end in common, annular outer waste channel (2), substantially concentric with channel (1).
- Each inlet opening (3) of the microchannel structures (K7-K12) may be used as an application area for reagents and samples.
- Each microchannel structure (K7-K12) is provided with a waste chamber (4) that opens into the outer waste channel (2).
- Each microchannel (K7-K12) forms a U-shaped volume-defining structure (7) and a U-shaped chamber (10) between its inlet opening (3) and the waste chamber (4).
- the normal desired flow direction is from the inlet opening (3) to the waste chamber (4) via the U-shaped volume-defining structure (7) and the U-shaped chamber (10).
- Flow can be driven by capillary action, pressure and centrifugal force, i.e.
- fluid can flow from the inlet opening (3) via an entrance port (6) into a volume-defining structure (7) and from there into a first arm of a U-shaped chamber (10).
- the volume-defining structure (7) is connected to a waste outlet for removing excess fluid, for example, radially extending waste channel (8) which waste channel (8) is preferably connected to the annular outer waste channel (2).
- the waste channel (8) preferably has a vent (9) that opens into open air via the top surface of the disk. Vent (9) is situated at the part of the waste channel (8) that is closest to the centre of the disc and prevents fluid in the waste channel (8) from being sucked back into the volume-defining structure (7).
- the chamber (10) has a first, inlet arm (10a) connected at its lower end to a base (10c) which is also connected to the lower end of a second, outlet arm (10b).
- the chamber (10) may have sections I, II, III, IV which have different depths, for example each section could be shallower than the preceding section in the direction towards the outlet end, or alternatively sections I and III could be shallower than sections II and IV, or vice versa.
- a restricted waste outlet (11), i.e. a narrow waste channel, is provided between the chamber (10) and the waste chamber (4). This makes the resistance to fluid flow through the chamber (10) greater than the resistance to fluid flow through the path that goes through volume-defining structure (7) and waste channel (8).
- the top and bottom surfaces of the waste chamber (4) are preferably separated by one or more supports (12) to ensure that the top and bottom surfaces of the microfluidic device do not bend inwards towards the waste chamber (4) and thereby change its volume.
- the volume-defining structure (7) is U-shaped with the entrance port (6) opening into the upper end (i.e. the end nearest to the centre of the disc) of one of the arms (7a) of the U and the waste channel (8) connected to the upper end of the other arm (7b) of the U.
- the vent (9) is also placed at the top of this other arm (7b).
- the base (7c) of the U-formed volume-defining structure (7) is connected to the upper end of a first arm (10a) of the chamber (10).
- an additional application area (13) that opens out into the top surface of the disc and is connected to the entrance port (6).
- This additional application area (13) can be used when it is desired to add different reagents or samples to each of the different microstructures (K7-K12).
- a hydrophobic break is preferably provided at the connection (16) of the chamber (10) to the volume-defining structure (7) in order to guide fluid into arm (7b)
- the outer annular waste channel (2) may be sectioned so as to collect waste from a selected number of closely located microchannel structures.
- Hydrophobic breaks can be introduced into the microchannel structures (K7-K12), for example by marking with an over-head pen (permanent ink) (Snowman pen, Japan), and suitable places for such breaks (shown by crosshatching in the figures) include: (a) between microchannel structure inlets (3) in the inner annular application channel (1), (b) each opening (15) into the outer annular waste channel (i.e. the openings of the waste chambers) and, (c) if present, also the radial waste channels (5) which connect the inner annular application channel (1) and the outer annular waste channel (2), and also the waste channel (8) which guides away excess fluid from the volume-defining structure (7).
- hydrophobic breaks The purpose of the hydrophobic breaks is to prevent capillary action from drawing the fluid into undesired directions. Hydrophobic breaks can be overcome by centrifugal force i.e. by spinning the disc at high speed.
- the sample to be analysed is in the form or cells or sedimenting material or particles then it can be held in the lower U-channel by a particle filter (21) (shown by a dotted line in figure 1b and 1d) or the flow through the chamber (10) can be controlled such that particles are retained in the chamber while fluids flow through it - as will be described later.
- a particle filter (21) shown by a dotted line in figure 1b and 1d
- the flow through the chamber (10) can be controlled such that particles are retained in the chamber while fluids flow through it - as will be described later.
- a first reagent or sample fluid X can be introduced into the chamber (10) by connecting a source (not shown) of the fluid X to the common annular inner application channel (1) from where it flows by capillary action and/or, if the disc is spun, centrifugal force to the lower U-bend. If the volume of fluid X which is introduced into common annular inner application channel (1) is in excess (i.e. is greater than the volume of the chamber (10) up to the level of the restricted channel (11) (distance L4 in figure 1d)) then some of it flows to waste via the radial waste channels (5 and 8) while the rest flows to waste chamber (4) via the chamber (10) though the restricted channel (11) as shown in figure 2.
- fluid Y When it is time to add a new reagent or sample fluid Y, then fluid Y is added by the common annular inner application channel (1) (or, alternatively, as shown in figure 3a) by the additional application area (13)).
- the fluid Y travels by capillary action through the volume-defining structure (7) and down the waste channels (5 and 8) as shown in figure 3a). It cannot flow into chamber (10) as the air cushion (19) contained between the base of the volume defining structure and the top of the fluid in arm (7a) of the chamber acts as a barrier to prevent the fluid flowing into chamber (10).
- an air cushion (19) can be left between the first fluid X and the second fluid Y by making the distance L4 from the base of the U-bend in the chamber (10) to the restricted channel (11) less than the distance L3 from the base of the U-bend in the chamber (10) to the base of the U-bend of the volume-defining structure (7).
- This can prevent the second fluid Y from flowing by capillary action into the chamber (10) and can also prevent mixing of the fluids X and Y.
- the vent (9) which is open to atmospheric pressure, makes it easier for the second fluid Y to flow towards the waste channel (2).
- Gentle, i.e. low speed, spinning of the disc (D) empties the excess fluid Y from waste channel (8), leaving the volume-defining structure (7) full of fluid Y, as shown in figure 3b.
- All of the first fluid X in the chamber (10) can be displaced by the second fluid Y by spinning the disc if the volume of the second fluid in the volume-defining structure (7) and any air between the first and second fluids is equal to or greater than the volume of the first fluid X in the chamber (10).
- This can be achieved by ensuring that the volume of the volume-defining structure (7) is greater than the volume of the chamber (10).
- This can be achieved by making the arms (7a) and (7b) of the volume-defining structure longer than the arms of the chamber (10), and/or by making the cross-sectional area of the arms of the volume-defining structure (7) greater than that of the arms of the chamber (10).
- Figure 4a shows an intermediate situation where the disc is being spun and centrifugal force causes fluid Y to flow from the volume-defining structure (7) into chamber (10), thereby displacing first fluid X which flows to waste via restricted channel (11). Any excess second fluid Y flows out of the chamber (10) through the restricted channel (11) into waste chamber (4).
- Figure 4b shows that the second fluid Y has replaced the first fluid X. This process can be repeated using different fluids as often as is desired.
- the chamber (10) In the event that the fluids contain particles and it is desired to hold them in the chamber it is possible to provide the chamber (10) with a particle filter (21) with suitable sized orifices. In the event that it is necessary to only temporarily hold the particles in the chamber (10) then the sections I, II, III, IV of the chamber (10) which have different depths can be used to temporarily trap the particles. This is done by increasing the speed of rotation of the disc so that the particles collect at the boundary wall between two sections while the fluid flows over the wall.
- particles can be selectively held in, or flushed out of a chamber (10'), which does not have a particle trap or sections having different depths as shown in figure 5. This can be achieved as follows:
- Particles that have been sedimented, or otherwise collected, in the bottom of the chamber (10') can be drawn out of the chamber (10') by the meniscus of a fluid which flows out of the chamber (10').
- the meniscus between the fluid in the chamber and the air cushion passes the particles they are entrained by the meniscus and flow out of the chamber.
- This can be achieved by choosing a suitably low rate of acceleration of the disc (known as "ramp speed"). If however it is desired to retain the particles in the chamber then it is necessary to ensure that the air cushion is not driven through the chamber (10') by the fluid in the volume-defining structure when the disc is spun.
- a suitably high rate of acceleration of the disc it is possible to cause the fluid in the volume-defining structure to flow down the sides of the channel, through the air cushion (19'), without displacing the air cushion (19').
- a ramp speed of up to 3500 rpm/s 2 transports the particles further in the channel system. With a ramp speed greater than 3500 rpm/s 2 the fluid/air interface (meniscus) does not enter the U-chamber and the air bubble stays still or moves in the opposite direction to the centrifugal force.
- the exact ramping speeds to achieve the desired effect are naturally dependent on the type of fluid used and are most suitably determined by experimentation.
- the arm (7b') of the volume-defining structure (7') is not connected to a waste channel (8), but is instead enlarged at its end nearest the centre of the disk in order to form a reservoir (61) for fluid to prevent fluid overflowing out of a vent (9').
- This vent and/or sample inlet (9') vents this reservoir (61) to atmosphere and can also permit samples to be introduced into the structure.
- the reservoir (61) preferably has a length which makes the length of the volume defining structure i.e. reservoir (61) and arm (7b') equal to or greater than the length of arm (7a').
- vent (9') is made so small that the surface tension of the fluid prevents it from flowing out of the vent when the volume-defining structure (7') is being charged by spinning, then the amount of fluid which can enter the volume-defining structure (7') is minimised and no fluid is wasted.
- the volume of the volume defining structure must be greater than the volume of the chamber (10). If the arm (10a) of the chamber is made to widen from its upper end to its lower end then it is possible to push the air barrier (19) out of the chamber when adding a second fluid without the two fluids mixing.
- All the chambers of the present invention can be provided with heating means in the form of a coating as shown crosshatched in figure 7.
- This coating (71) which can be painted or printed or applied in some other way to one or both sides of the disk in the vicinity of the chamber, can absorb energy from electromagnetic radiation which is directed onto it and thereby heat up the chamber.
- the incident radiation can be infra red light, laser light, visible light, ultraviolet light, microwaves or any other suitable type of radiation.
- the heating up of the chamber can be used to initiate or accelerate reactions in the chamber. If the disk is stationary while the chamber is being heated then if the fluid boils it will produce bubbles of vapour which will travel up the arms of the chamber and may even pass out into the waste channel (8) and waste chamber (4).
- the fluid should remain in the chamber after the heating has been finished.
- This can be achieved in the present invention by spinning the disk at the same time that radiation is incident on the coating (71).
- the radiation sources (not shown) can be focused onto areas that the coating passes through as the disc spins.
- the coating can be dimensioned such that heat is only applied to only the smallest amount of the base consistent with adequate heating of the reagents. In this way the arms of the U are keep cool and provide condensation surfaces for the fluid vapour to condense on. The centrifugal force exerted on the condensed vapour causes it to flow back into the base of the chamber.
- each further chamber may have a plurality of inlets and a plurality of outlets so that samples and reagents may be combined in a chamber.
- the subsequent results of any process, which has taken place in a chamber, can be dispensed to one or more additional chambers for further processing or sent to the waste channel.
- An example of this is shown in figure 8.
- Figure 8 shows a microstructure, of a design similar to that shown in figure 6, in which the base (110c) of U-shaped chamber (110) is connected by a base outlet channel (134) to a second chamber (136), which second chamber (136) is positioned further away from the centre of the disk than second chamber (110).
- Second chamber (136) is vented to atmosphere by a vent (138) that opens out on the top surface of the disc.
- Second chamber (136) is also provided with an inlet/outlet connection (140) that also opens out on the top surface of the disk.
- Inlet/outlet (140) can be used to supply substances to second chamber (136) e.g. by injecting them into connection (140) and/or to extract substances from second chamber (136) e.g.
- hydrophobic break (132) is dimensioned so that when the disc is spun at a certain number of revolutions per second then any fluid in chamber (110) leaves the chamber via chamber outlet arm (110b), and when the disc is spun at a higher number of revolutions per minutes then the centrifugal force acting on the fluid is sufficient to overcome the hydrophobic effect of hydrophobic break (132) and the fluid flows into second chamber (136).
- the outlet arm (110b) of chamber (110) is almost as long as inlet arm (1 10a).
- the level of fluid in inlet arm (110b) will be very close to the base (107c') of the volume-defining structure (107').
- a second fluid is supplied to the volume-defining structure (107'), e.g. via inlet (109') in the reservoir (161), it will come into direct contact with the first fluid in the chamber (110) and no air bubble will form between the two fluids.
- This arrangement can be used to facilitate mixing of two fluids.
Abstract
Claims (10)
- Microstructure pour fluides disposée dans un disque rotatif (D), caractérisée en ce qu'elle comprend une structure définissant un volume en forme de U (7, 107), comprenant :. un premier bras (7a) relié à son extrémité supérieure ou au voisinage de celle-ci à un orifice d'entrée (6), dans lequel l'extrémité inférieure dudit premier bras (7a) est plus éloignée, ou à la même distance, du centre dudit disque (D) que ledit orifice d'entrée (6) ;. un deuxième bras (7b) relié à son extrémité supérieure ou au voisinage de celle-ci à un premier canal d'évacuation (8) ou évacuation ou orifice d'entrée d'échantillon (9', 109'), par l'intermédiaire d'un réservoir (61, 161) pour fluide, dans lequel ledit canal d'évacuation (8), s'il est présent, est plus éloigné du centre dudit disque (D) que ledit orifice d'entrée (6) ; et. une base (7c) positionnée plus loin dudit centre dudit disque (D) que lesdits premier et deuxième bras (7a, 7b), cette base (7c) reliant les extrémités inférieures desdits premier et deuxième bras (7a, 7b), ladite base (7c) étant reliée à un bras d'entrée (10a, 110a) d'une chambre en forme de U (10, 110), à l'extrémité supérieure dudit bras d'entrée (10a) ou au voisinage de celle-ci, ladite chambre en forme de U (10, 110) comprenant de plus :. une base (10c, 110c) et un bras de sortie (10b, 110b), ladite base (10c, 110c) reliant l'extrémité inférieure dudit bras d'entrée (10a, 110a) à l'extrémité inférieure dudit bras de sortie (10b, 110b), et ledit bras de sortie (10b, 110b) étant relié, à son extrémité supérieure ou au voisinage de celle-ci, à un deuxième orifice de sortie d'évacuation (11), et ladite base (10c, 110c) étant plus éloignée, ou à la même distance, du centre dudit disque (D) que les extrémités inférieures desdits bras d'entrée et de sortie (10a, 10b ; 110a, 110b) de ladite chambre en forme de U (10, 110).
- Microstructure selon la revendication 1, caractérisée en ce que ledit premier canal d'évacuation (8) est muni d'une évacuation (9).
- Microstructure selon la revendication 1, caractérisée en ce que le deuxième bras (7b) est relié à l'évacuation ou à l'orifice d'entrée d'échantillon (9', 109').
- Microstructure selon l'une quelconque des revendications 1 à 3, caractérisée en ce que la résistance à l'écoulement de fluide à travers ledit deuxième orifice de sortie d'évacuation (11) est supérieure à la résistance à l'écoulement de fluide à travers ledit premier canal d'évacuation (8).
- Microstructure selon l'une quelconque des revendications précédentes, caractérisée en ce que la longueur de la première structure définissant un volume en forme de U (7, 107) est supérieure à la longueur de la deuxième structure de la chambre en forme de U (10, 110).
- Microstructure selon l'une quelconque des revendications précédentes, caractérisée en ce que ladite structure de chambre (10, 110) est au moins partiellement recouverte par un revêtement (71), qui peut absorber l'énergie d'un rayonnement électromagnétique qui est dirigé sur celui-ci, et, par conséquent, chauffer ladite structure de chambre (10).
- Microstructure selon l'une quelconque des revendications précédentes, caractérisée en ce que ladite structure de chambre (10, 110) comporte des sections I, II, III, IV qui ont des profondeurs différentes et qui peuvent être utilisées pour piéger et libérer des matériaux de sédimentation ou d'autres particules .
- Microstructure selon l'une quelconque des revendications précédentes, caractérisée en ce que ladite structure de chambre (110) est reliée par sa base (110c) à une deuxième chambre (136) positionnée plus loin dudit centre dudit disque (D) que ladite structure de chambre (110) à l'aide d'un canal (134), dans lequel se trouve une interruption hydrophobe (132) positionnée à la jonction (130) entre la chambre (110) et le canal (132) ou au voisinage de celle-ci.
- Utilisation d'une microstructure dans un disque rotatif (D) selon l'une quelconque des revendications précédentes, pour distribuer des volumes prédéterminés de fluide à une chambre (10, 110) dans ledit disque rotatif.
- Procédé pour remplacer un fluide original dans une chambre (10, 110) dans un disque rotatif (D), caractérisé par les étapes consistant à :. disposer une microstructure selon l'une quelconque des revendications 1 à 8, contenant ledit fluide original dans une chambre (10, 110) ;. remplir ladite structure définissant un volume (7) par un fluide de remplacement ; et. faire tourner ledit disque (D) à une vitesse suffisamment élevée pour que ledit fluide de remplacement se déplace sous l'effet de la force centrifuge vers l'intérieur de ladite chambre, tandis qu'en même temps, le fluide original dans la chambre (10) est forcé hors de la chambre par le fluide de remplacement entrant.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP00992452A EP1239962B1 (fr) | 1999-12-23 | 2000-12-22 | Dispositif de microanalyse |
Applications Claiming Priority (6)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
WOPCT/EP99/10347 | 1999-12-23 | ||
PCT/EP1999/010347 WO2000040750A1 (fr) | 1998-12-30 | 1999-12-23 | Procede de sequençage d'adn a l'aide d'un dispositif microfluidique |
SE0001779A SE0001779D0 (sv) | 2000-05-12 | 2000-05-12 | Microanalysis device |
SE0001779 | 2000-05-12 | ||
PCT/EP2000/013145 WO2001046465A2 (fr) | 1999-12-23 | 2000-12-22 | Dispositif de microanalyse |
EP00992452A EP1239962B1 (fr) | 1999-12-23 | 2000-12-22 | Dispositif de microanalyse |
Publications (2)
Publication Number | Publication Date |
---|---|
EP1239962A2 EP1239962A2 (fr) | 2002-09-18 |
EP1239962B1 true EP1239962B1 (fr) | 2007-07-18 |
Family
ID=27222880
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP00992452A Expired - Lifetime EP1239962B1 (fr) | 1999-12-23 | 2000-12-22 | Dispositif de microanalyse |
Country Status (1)
Country | Link |
---|---|
EP (1) | EP1239962B1 (fr) |
-
2000
- 2000-12-22 EP EP00992452A patent/EP1239962B1/fr not_active Expired - Lifetime
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Publication number | Publication date |
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EP1239962A2 (fr) | 2002-09-18 |
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