The present application is a 371 of International application PCT/EP2017/062602, filed May 24, 2017, which claims priority of EP 161 77 162.1, filed Jun. 30, 2016, the priority of these applications is hereby claimed and these applications are incorporated herein by reference.
The invention relates to a flow cell having at least one reservoir region containing a reagent.
As is known, microfluidic flow cells are increasingly employed in diagnostics, analytics and or synthesis of substances, primarily in Life Sciences. As is known, such flow cells often process very small volumes of reagents, which interact with the samples to be analyzed or processed and which have to be introduced into the flow cells in the course of production or during usage of the flow cells.
Reagents can be stored within the flow cells in storage spaces, transport channels or containers introduced into the flow cells. For the storage of liquid reagents, blisters which are closed off by predetermined breaking point barriers and which are preferably produced from aluminum laminates can in particular be considered. The holding capacity of such blisters can neither be reduced nor enlarged as desired. In particular large blisters require a cover housing which protects against accidental squeezing. In the downward direction, the holding capacity is limited by production tolerances, wherein a lower limit is around 50 microliters.
In the case of storage spaces integrated in the flow cell, although such limitations do not exist, complex connecting channels are necessary for filling and venting, which, following placement of the reagent within the flow cell, then have to be sealed by welding or bonding in order to close off the storage space in a hermetic and storage-stable manner. Liquid reagents can be, for example, fluorescent dyes, acids, alkalis, alcohols, bead solutions, lysis buffers, antibodies, enzymes, DNA fragments, PCR reagent mixtures or wash buffers.
The object of the invention is to provide a new flow cell having a reservoir region for small liquid reagent volumes, which flow cell is producible with reduced complexity in relation to the prior art.
The flow cell according to the invention which achieves this object is characterized in that the reservoir region is delimited by a carrier element introduced jointly with the reagent into an opening in the flow cell, wherein the carrier element closes off the reservoir region to the outside in a fluid-tight manner and has a vessel and/or capillary structure holding the liquid reagent on the carrier element.
Advantageously, by virtue of the present invention, both in the course of production and during usage of the flow cell, a small volume of a liquid reagent can be introduced into the flow cell, preferably reagent volumes between 1 and 100 microliters, in particular between 5 and 50 microliters. Complex venting channels which have to be sealed are able to be avoided. The reagent to be stored can be comfortably applied to the carrier element, into the vessel and/or capillary structure of the carrier element outside the flow cell, by pipetting or dipping.
In one embodiment of the invention, the reservoir region within the flow cell is hermetically closed off against inner cavities of the flow cell by at least one predetermined breaking point barrier. In this way, the flow cell provided with the liquid reagent is able to be stored on a long-term basis.
The carrier element can be connected to the flow cell solely by force closure and/or form closure, for example when the liquid reagent, during usage of the flow cell, is introduced into the flow cell. Alternatively or additionally, the flow cell is welded and/or bonded to the flow cell in a connecting region arranged at a distance from the reagent. As a result of the distance of the connecting region from the reagent, impairments of the reagent as a result of welding heat or adhesive fumes, can be avoided.
In a particularly preferred embodiment of the invention, the reservoir region is fluidically connected to at least one transport channel of the flow cell.
In particular, one transport channel of the flow cell leads toward the reservoir region and one transport channel of the flow cell away from the reservoir region, wherein in the channel or each of the channels, a predetermined breaking point barrier, which hermetically encloses the reagent, can be formed.
The opening is preferably formed in a plate-like substrate of the flow cell, and the flow cell comprises a cover, in particular a cover foil, which is connected to the substrate and which covers the opening and, where appropriate, the at least one transport channel.
The reservoir region can be delimited within the flow cell alone by the vessel and/or capillary structure of the carrier element or by the vessel and capillary structure and the cover.
Alternatively, the reagent adjoins with a free liquid surface an interior of a chamber, in particular mixing chamber, formed in the flow cell.
The carrier element is preferably configured in the form of a stopper filling the opening and comprising a front side having the vessel and/or capillary structure. In particular, the carrier element has a conical portion, which can ensure a seal-tight closure of the reservoir region given sufficient venting of the reservoir region.
Expediently, the carrier element, on an outer side facing away from the reservoir region, is provided with handling devices and comprises, in particular, a seat for connection to an assembly tool. The handling devices can be useful both in the filling of the vessel and/or capillary structure and in the fitting of the carrier element containing the reagent.
In a further embodiment, the carrier element, on an outer side facing away from the reservoir region, has a collar, which forms the above-stated connecting region and via which a welding and/or bonding to the flow cell can be realized.
In a further embodiment, the vessel and/or capillary structure has a groove which receives the reagent or a channel which receives the reagent, wherein the groove or channel is preferably at at least one end open to a peripheral surface of the carrier element.
In a particularly preferred embodiment of the invention, devices for detaching the liquid reagent from the vessel and/or capillary structure are provided.
Such devices can be designed to detach the reagent by a fluid which rinses off the reagent or by an inertia force, in particular centrifugal force, which detaches the reagent. For the generation of a centrifugal force, the flow cell can be set in rotation during use, for example by an operator device.
If the reagent, with a free liquid surface, adjoins an interior of a mixing chamber formed in the flow cell, a fluid provided in the mixing chamber can wash off the liquid reagent, in particular by shaking of the flow cell. Alternatively, in the mixing chamber the liquid reagent can be washed off by a single or multiple flushing as a sample liquid or another mixing or rinsing liquid moves back and forth.
In a particularly preferred embodiment of the invention, the groove or channel of the vessel and/or capillary structure is aligned with the transport channel leading toward the reservoir region and leading away from the reservoir region, so that a rinsing flow can flow through the reservoir region.
In a further preferred embodiment of the invention, the transport channel leading toward the reservoir region and the transport channel leading away from the reservoir region are connected by a bypass which circumvents the reservoir region. Air which is present between the liquid reagent and a rinsing flow can thus flow past the reservoir region. If the flow cross section of the bypass is smaller than that of the reservoir region, the reagent is fully washed out with the rinsing fluid.
In a further embodiment, the flow cross section of the reservoir region is smaller than the flow cross section of the transport channel leading toward and/or leading away from the reservoir region.
Furthermore, the flow cross section of the bypass can also be larger than the flow cross section of the reservoir region, so that a possibly desired delayed or gradual rinsing-out over a longer period is realized.
The carrier element can be rotatably connected to the flow cell and have, for example, a stop by which the above-stated alignment of the reservoir region with the channels is assured.
In a further embodiment of the invention, at least the vessel and/or capillary structure of the carrier element has a hydrophilic surface, by which, when wetted with the liquid reagent, a desired reagent volume is able to be more precisely measured.
For the further refinement of the measurement, the vessel and/or capillary structure of the carrier element can further be adjoined by a hydrophobic surface of the carrier element in order to achieve a sharp contrast between wettability and non-wettability.
Naturally, a carrier element could also form a plurality of reservoir regions within a flow cell.
The invention is further explained below with reference to illustrative embodiments and the attached drawings relating to these illustrative embodiments, wherein:
A flow cell represented in part in
The transport channel 3 opens out into a through opening 4 closed at one end by the foil 2 and having a conical portion 5. The latter is lengthened by an annular protrusion 6 connected to the substrate 1. Lying diametrically opposite the mouth of the transport channel 3 is a mouth of a further transport channel, which latter is not visible in
A carrier element 7 for a liquid reagent 8 can be inserted into the through opening 4. The carrier element 7, which is rotationally symmetrical in the illustrative embodiment shown, has a peripheral surface 9 corresponding to the through opening 4 and is provided on an outer side with a circumferential collar 10. A depression 11 opening out to the outer face of the carrier element 7 serves as a seat for receiving a handling tool.
On its front side facing away from the outer face, the carrier element 7 has a vessel and/or capillary structure in the form of a groove 12 as can be seen with reference to
Prior to the fitting of the flow cell, the liquid reagent 8 is applied, for example by pipetting or immersion of the carrier element into a reagent supply, to the carrier element 7, where it is held in the groove 12 by capillary forces. Also following introduction of the carrier element 7 into the through opening 4 and welding and/or bonding of the collar 10 to the annular protrusion 6, the liquid reagent 8 initially remains in the groove 12 covered by the foil 2, which groove, within the now finished flow cell, forms, together with the foil 2 to which the carrier element 7 reaches, a reservoir region 13.
The storable liquid volume of such a reservoir region 13 lies between 1 and 100 microliters, preferably between 2 and 20 microliters.
The substrate 1 and the covering foil 2 preferably consist of a plastic, in particular the same plastic, for example PMMA, PC, COC, COP, PP or PE. For the preferably injection molded carrier element, in particular COC, PP, PET, PE, PMMA, PC, PEEK, TPE or silicone enter into consideration as the plastic. The carrier element 7 too can consist of the same plastics material as the substrate 1 and/or the covering foil 2. The substrate preferably consists of a more brittle plastic, such as PC or COC, the carrier element 7 of a more ductile material, such as PE or PP, in order to make the conical compression joint more pressure-stable.
During use of the flow cell, the liquid reagent 8, when necessary, is removed from the reservoir region 13, for example by a further fluid that flows in via the transport channel 3, for example a sample to be analyzed or a further stored reagent, for example a wash buffer or dilution buffer. The further fluid forces the liquid reagent 8 out of the reservoir region 13 aligned with the channel 3 into the aforementioned, diametrically opposite transport channel and can mix there with the stored reagent.
If the flushing-out and displacement of the liquid reagent 8 from the reservoir region 13 itself is realized by a liquid, then the formation of an air cushion between the liquid reagent and the latter liquid must as far as possible be avoided. A bypass 14, which, according to
As shown by
Air streaming ahead of a flushing-out liquid flows through the bypass 14 or 14′, while the liquid reagent initially continues to be held in the reservoir region 13 by capillary forces. Once the flushing liquid reaches the reservoir region, then also the bypass 14, 14′ fills with flushing liquid. Since the flow cross section of the bypass 14, 14′ is smaller, however, than the flow cross section in the reservoir region 13, a lower flow resistance is obtained in the reservoir region 13 and the flushing liquid transports the liquid reagent 8 out of the reservoir region.
The inflow or outflow channel is preferably aligned with the groove 12 forming the vessel and/or capillary structure, wherein the cross sections preferably have a width of 0.05 to 2 mm and a height of 0.1 to 3 mm.
At variance with the shown examples, bypasses could also be formed by virtue of the fact that the cover foil 2 is not fixedly connected to the substrate right up to the rim of the through opening 4 and is deflectable by external means, for example by underpressure, in order to form vent slots.
The flow cross section of lateral vent slots, as are shown in
In a further embodiment, the reservoir region can be smaller in cross section than the cross section of the transport channels fluidically connected to the reservoir region, as is indicated in
Further illustrative embodiments of carrier elements emerge from
In
According to
As in the illustrative embodiment of
A detail of a flow cell having a channel region 23 and two mixing chambers 19′, 19″ is shown by
A predetermined breaking point barrier 26 is found between the reservoir region 13 and the mixing chamber 25 of the flow cell of
A flow cell shown in
Changes to the surface properties of the plastic forming the carrier element can be made (hydrophilically or hydrophobically), using wet chemical methods, by application of wetting agents or surfactants and subsequent drying (hydrophilic or hydrophobic). In addition, a surface activation can be performed by means of plasma, flame treatment or corona treatment (hydrophilic). Surface coatings by plasma polymerization, for example vitreous layers, hydrophilically or hydrophobically, or combinations thereof, can be applied all over/in full, or in a locally masked manner.
Instead of the hydrophilization coating applied, in
A flow cell shown in part in
The pressure source conveys the fluid with the rinsed-off reagent into the mixing chamber 45, in which, by compression of air contained therein, a counterpressure to the pressure source builds up. The pressure of the pressure source is variable, so that, as a result of the counterpressure built up in the mixing chamber 45, a reversal of the motion of the fluid with the rinsed-off reagent can be achieved, and the fluid with the rinsed-off reagent can be moved back and forth, with intensive intermixing, by variation of the pressure of the pressure source.
A flow cell represented in part in
By actuation of the blister 46, the fluid presses against the predetermined breaking point barrier 47 and opens up the predetermined breaking point barrier 47. When the valve 48 is closed, the fluid with the rinsed-off reagent is conveyed into the storage chamber 49, in which a counterpressure builds up. The counterpressure can be used for a return transport of the fluid with the rinsed-off reagent into the blister 46, wherein the wall of the blister inflates again. By repeated actuation of the blister 46, the fluid with the rinsed-off reagent is moved back and forth with intensive intermixing. Via the opened valve 49, the mixture can then be transported away for further use within the flow cell.
In the flow cells described above with reference to
By way of supplement, it should further be mentioned that a vessel and/or capillary structure also be formed merely by hydrophilized carrier surface, in particular circular carrier surface, which, where appropriate, is adjoined by a hydrophobic surface.
Number | Date | Country | Kind |
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16177162 | Jun 2016 | EP | regional |
Filing Document | Filing Date | Country | Kind |
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PCT/EP2017/062602 | 5/24/2017 | WO |
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WO2018/001647 | 1/4/2018 | WO | A |
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