EP1343587A2 - Device for receiving and discharging liquid substances - Google Patents

Abstract

The invention relates to a device for controlling the amount of liquid substances received and discharged. The aim of the invention is to produce a device, enabling a plurality of different liquid substances to be received and discharged from micro or nano titer plates. According to the invention, it is possible to carry out one or more chemical or biological reactions in said device and to receive liquid substances with differing viscosities, as a result of capillary channels (1) which are arranged at an equal distance from each other and are provided in a row, said channels being brought together in a communicative link with a chamber (2) which is impinged upon by high and low pressure, whereby the capillary channels (1) are embedded in the plate (3) and one sieve-type membrane (4) is associated with the ends of the capillary channels (11) on the inner side of the pressure chamber. According to the invention, an area (5) is provided above each end of the capillary channels for receiving a liquid substance. Said areas (5) are arranged separately from each other and are disposed inside the chamber (2) which can be impinged upon by high and low pressure.

Description

Device for receiving and dispensing liquid substances

description

The invention relates to a device for the controlled uptake and release of liquid substances, preferably for the uptake and release from or into micro- and nanotiter plates, wherein at the same time preferably all cavities of at least one row of a titer plate can be detected. In the macroscopic range, suction lifters for holding larger volumes are known, in which pipettes are provided at the same distance from each other in the form of a rake, all pipettes being connected together with a balloon, piston or similar assembly that can be pressurized with overpressure or underpressure, so that at the same time, several a liquid substance can be removed from adjacent containers.

Also known is a reactor for microchemical and / or microbiological reactions, which comprises a pipette with a dosing approach (DE 196 42 777 AI), a reactive solid phase support with at least one immobilized reaction partner being provided in the pipette near its lower, narrowed end is. Such designs, as also described in DE 197 23 469 AI, cannot be miniaturized for mechanical reasons and are also not the subject of the present invention.

The surface of this elevation is preferably hydrophobic or solvent-repellent on the outside and inside by chemical coating or by a physical structuring or combinations of both (e.g. a self-cleaning surface "seif cleaning surface").

The invention is based on the object of specifying a device with the aid of which a large number of different liquid substances can also be simultaneously absorbed from microplates or nanotiter plates, can be dispensed again in the same grid in defined partial volumes if required, and the possibility of carrying out one or more chemical substances or biological reactions in the device itself and a simultaneous absorption of liquid substances of different viscosity should be possible.

The object is achieved by the characterizing features of the first claim. Advantageous configurations are covered by the subordinate claims.

The essence of the invention is that at least in one row, equally spaced capillary cannulas are provided, which are brought into a communicating connection with a chamber to which an overpressure or underpressure can be applied, the capillary cannulas being embedded in a plate and at least the capillary cannula ends on the inside of the pressure chamber a sieve-like membrane is assigned and above each capillary cannula end there is a space for the absorption of a liquid substance, within which there is the possibility of carrying out chemical or biological reactions, the spaces being separated from one another and all the spaces together within those that can be pressurized with overpressure or underpressure Are arranged chamber, wherein the capillary cannulas are designed so that their internal volume is smaller than the volume that can be absorbed by each capillary.

The present invention permits a highly parallel liquid transfer and the removal of substance, in particular from micro or Namotiter plates of comparable design or comparable containers.

The proposed device can be used both for solid-phase-coupled syntheses and in particular for liquid-phase syntheses in all variants. The decisive advantage of the device is that defined volumes of different viscosities can be simultaneously absorbed and dispensed, a highly parallel and efficient liquid transfer being ensured.

In particular, transfer operations for bioassay applications can be carried out effectively with this device, for example dilution series of library substances in the assay. This means that bioassays can also be carried out after washing, introducing the target substance into solution and after photo-cleaving the synthesized library substances. Another possible use is the removal of substances for further analysis processes by transferring them into corresponding analysis vessels. The integration of bioassay and synthesis made possible by the device also enables software-supported evaluation.

The invention will be explained in more detail below with the aid of schematic exemplary embodiments. Show it:

Figure 1 shows a possible embodiment of the device for receiving and dispensing liquid substances in a longitudinal section.

2a shows a first possible embodiment of parts of the device which are essential to the invention,

2b shows a second possible embodiment of parts of the device essential to the invention, FIG. 2c shows a third possible option for parts of the device essential to the invention,

Fig. 3 shows a preferred embodiment of the device with the formation of spaces of defined volume within which reactions can be carried out; and Fig. 4 shows, by way of example, a lower view of the device, which is provided with cannulas in several rows and columns, the mutual spacing of which is the cavity grid of a titer plate which can be put in place equivalent.

Figure 1 shows schematically a possible embodiment of the device in a longitudinal section. Capillary cannulas 1, which are equally spaced from one another, are provided in a row, which are brought into a communicating connection together with a chamber 2, which can be pressurized with an overpressure or negative pressure via a connector 8, the capillary cannulas 1, which are in particular designed as steel cannulas, which are inserted into a Plate 3 are inserted and fixed by gluing. At least the capillary cannula ends 11 on the inside of the pressure chamber, which in the example are flush with the plane of the plate 3 on the pressure chamber side, is initially assigned a sieve-like membrane 4, above which each capillary cannula end is provided with a space 5 for holding a liquid substance, the spaces 5 being separated from one another and all rooms 5 are arranged together within the chamber 2 which can be pressurized with an overpressure or underpressure, which ensures that all capillary cannulas 1 can be pressurized with an identical pressure. Depending on the design, ie depending on the size of the perforations provided for the sieve-like membranes 4, it can be achieved in the example according to FIG. 1 that a defined volume presented to the respective capillary cannulas can be drawn into the rooms 5 with sufficient negative pressure, or with a sufficiently small perforation of the sieve-like membranes 4, depending on the surface tension of the respective liquid substances, only the capillary cannulas 1 to the membrane 4 are filled only from any given volume, because the sieve-like membranes 4 prevent the liquid substances from passing through. In the latter case, a defined volume of the liquid substances is also taken up again, with the advantage here that substances of different viscosities can be taken up simultaneously with one and the same device.

In the example, each room 5, if it is to be used as a reaction room, should have a volume of 10 nl to 2 μl. The assigned capillaries should have a volume of 1 nl to 120 nl. Depending on the specific circumstances, any other dimensions with regard to the volumes mentioned are possible, however, in the event that 5 reactions are to be carried out in the rooms, it must be ensured that the respective rooms 5 have a larger volume than the capillary cannulas assigned to them to prevent crosstalk of reaction liquids between rooms 5.

FIG. 2a shows a first embodiment of parts of the device shown in FIG. 1 that are essential to the invention. In this example, the spaces 5 mentioned and the sieve-like membranes 4 are through one-piece component 6 is formed, into which a plurality of recesses for forming the rooms 5 are made down to a floor area 61 and the respective remaining floor areas are provided with a perforation 62.

FIG. 2b shows a second embodiment of parts of the device shown in FIG. 1 which are essential to the invention. In this example, the spaces 5 are formed by a component 6, into which a plurality of continuous recesses 63 are initially made, which are provided with separate sieve-like membranes 4 on the sides facing the capillary cannulas 1.

Finally, FIG. 2c shows a third embodiment of parts of the device shown in FIG. 1 which are essential to the invention. In this example, the sieve-like membrane 4 is formed by a continuous membrane which covers all the recesses 63 and is introduced as a network between the plate 3 and the component 6 by gluing.

A preferred embodiment of the device with the formation of spaces 5 of defined volume, within which reactions can be carried out, is shown in FIG. In this case, two one-piece silicon or glass wafers 6a and 6b are provided, in each of which recesses are made congruent down to the bottom area and each have sieve-like membranes 4; 7 are introduced into the remaining floor areas by selective etching, which is known per se and therefore cannot be explained further here. The wafers 6a and 6b thus produced are attached to the sieve-like membranes 4; 7 opposite side connected to each other by anodic bonding, gluing or other joining techniques, which are also common methods that require no further explanation here. Other manufacturing processes for manufacturing the embodiments corresponding to the wafers 6a, 6b described above, such as structures formed in PE, by means of which sieves with openings between 0.5 μm and 35 μm and chamber sizes of the reaction spaces 5 between 10 nl and 8 μl can also be realized are also within the scope of the invention. It is essential in the example described above that the space 5 provided above each capillary cannula end and formed by the recesses in the two wafers 6a, 6b on the side facing the pressure chamber 2 (not shown in FIG. 3) is gas-permeable by a second sieve-like membrane 7 is closed, the perforations of this second membrane 7, depending on the surface tension of the liquid substance used and the size of an applied negative pressure, as far as smaller than that of the sieve-like membrane 4 assigned to the capillary cannula ends 11 of the pressure chamber, that a liquid passage through the second sieve-like membrane 7 is prevented. In the example, the sieve-like membrane 4 is given a sieve hole width of 10 μm and the membrane 7 has a sieve hole width of 1 μm. The respective rooms 5, in each of which a reaction is to be carried out, have a volume of 1 μl in the example and the capillary cannulas 1 have a volume of 100 nl.

A liquid reaction can be carried out, for example, as follows: Five different dissolved reagents are each taken up in succession with the device by an appropriately set negative pressure and drawn through the capillary-side sieve 4, so that the volume remains in the reaction space. Any further volume taken up is mixed with that in the reaction space during this process. The more fine-pored membrane 7 prevents the liquid from entering the chamber 2. Depending on the materials used for the device, reactions can also be carried out at elevated temperature.

Furthermore, it is within the scope of the invention to design the chamber 2 which can be subjected to an overpressure or underpressure to be detachable from the plate 3 (as indicated in FIG. 1) or at least openable above the recesses 63 (not shown in more detail) in order to provide a second access option to create the rooms 5, so that the rooms 5, in the case of designs according to FIGS. 1 and 2a to 2c from above, can be filled with an agent, if appropriate, with a second device designed analogously to the device described. Likewise, with the provided opening possibility of the chamber 2, a flooding of the rooms 5 can be realized.

If the above special description refers to sieve-like membranes, this also includes structures with irregularly distributed openings or passages, such as, for example,

Frits, so they perform the same function as the sieve-like membranes.

In extreme cases, the function of said membranes could in each case also be realized through a single, sufficiently small hole, but this is not necessarily a preferred embodiment.

In the context of the invention, it is advantageous to cover the walls of the spaces 5, at least the inner walls of capillary cannulas 1 and the surfaces of the sieve-like membranes 4; 7 to be provided with a hydrophobic surface and / or a self-cleaning physical microstructure (lotus effect) or with a solvent-repellent detergent, which facilitates the required cleaning of the device.

Finally, FIG. 4 shows an example of a lower view of a device, a plurality of capillary cannulas 1, which are equally spaced from one another in rows Z and columns Sp, being introduced into the plate 3 in a matrix-like manner and corresponding to the cavity distribution of a titer plate, not shown here. 4 space 5, not shown, for the absorption of a liquid substance.

Bezuεszeichenliste

1 capillary cannula

11 - Capillary cannula ends

2 (pressure) chamber

3 plate

4, 7 - sieve-like membranes

5 (reaction) room

6 one-piece component

61 - Floor area of the building menu 6

62 - perforation in the floor area 61

63 - continuous recesses

8 sockets

Z lines

Sp - columns

Claims

claims 1. Device for receiving and dispensing liquid substances, at least in one line equally spaced from each other Capillary cannulas (1) are provided, which are brought into a communicating connection together with a chamber (2) which can be pressurized with overpressure or underpressure, characterized in that. that the capillary cannulas (1) are embedded in a plate (3) and at least the capillary cannula ends (11) inside the pressure chamber are assigned a sieve-like membrane (4) and above each A capillary cannula end is provided in each case with a space (5) for holding a liquid substance, the spaces (5) being separated from one another and all spaces (5) being arranged together within the chamber (2) which can be pressurized or suppressed. 2. Device according to claim 1, characterized in that said spaces (5) and the sieve-like membranes (4) by a one-piece Component (6) is formed, in which several recesses are made down to a bottom area (61) and the respective remaining bottom areas are provided with a perforation (62). 3. Apparatus according to claim 1, characterized in that said spaces (5) are formed by a component (6), in which a plurality of continuous recesses (63) are made, on the side facing the capillary cannulas (1) with the sieve-like membrane (4) are provided. 4. The device according to claim 3, characterized in that the sieve-like membrane (4) is formed by a continuous membrane, all recesses (63) together. 5. The device according to claim 2, characterized in that the one-piece component (6) is formed by a silicon or glass wafer, in which the recesses are introduced into the bottom region and the sieve-like membranes (4) are introduced by selective etching, the remaining bottom regions. 6. The device according to claim 1, characterized in that the space provided above each capillary cannula end (5) on the pressure chamber (2) facing side of a second screen-like Membrane (7) is closed in a gas-permeable manner, the perforations of this second membrane (7), depending on the surface tension of the liquid substance used and the size of an applied negative pressure, so far smaller than that of the sieve-like membrane associated with the capillary cannula ends (11) on the inner side of the cannula chamber (4) that a liquid passage through the second sieve-like membrane (7) is prevented. 7. The device according to claim 6, characterized in that said spaces (4) are formed by two one-piece silicon or glass wafers (6a, 6b), introduced into the congruent recesses into the bottom area and the sieve-like membranes (4; 7) through selective etching are introduced into the remaining bottom areas and these wafers are connected to one another on the side opposite the sieve-like membranes (4; 7) by anodic bonding, gluing or other joining techniques. 8. The device according to claim 1, characterized in that the chamber with an overpressure or underpressure (2) from the plate (3) is designed to be detachable or at least above the recesses (63) to be opened in order to provide a second access to the rooms (5 ) to accomplish. 9. The device according to claim 1, characterized in that the walls of the spaces (5), the capillary cannulas (1) and the surfaces of the sieve-like membranes (4; 7) with a hydrophobic surface and / or a self-cleaning physical microstructure (lotus effect) or are provided with a solvent-repellent detergent. 10. The device according to claim 1, characterized in that the capillary cannulas (1) are formed by steel cannulas glued into the plate (3). 11. The device according to claim 1, characterized in that in the plate (3) matrix-like and the cavity distribution of a submitted Corresponding to the titer plate, a plurality of capillary cannulas (1) which are equally spaced from one another in rows (Z) and columns (Sp) are introduced, each of which is assigned a space (5) for the absorption of a liquid substance. 12. The apparatus according to claim 1, characterized in that each capillary cannula (1) is designed so that its internal volume is smaller than the volume that can be absorbed by the space (5) assigned to it.