WO2005116183A2

WO2005116183A2 - Reaction vessel, the production and the use thereof

Info

Publication number: WO2005116183A2
Application number: PCT/EP2005/005102
Authority: WO
Inventors: Helmut Herz; Klaus Kaufmann
Original assignee: Thermo Electron (Oberschleissheim) Gmbh
Priority date: 2004-05-17
Filing date: 2005-05-11
Publication date: 2005-12-08
Also published as: WO2005116183A3; DE102004024350A1

Abstract

The invention relates to a reaction vessel, the production thereof by an injection moulding method and to the use of said vessel for a bioreactor, wherein the inventive reaction vessel comprises at least one spring element which is placed on the external surface thereof and acts in the direction of the main axis of said vessel. Said spring element is also connected in a single piece to the vessel wall, thereby making it possible to carry out the mass production thereof in the form of a single-use product. Said vessel is provided with at least one optical lens disposed in the wall thereof which is translucent at least in certain areas.

Description

REACTION TUBE AND THE PRODUCTION AND USE THEREOF

The invention relates to a reaction vessel with a vessel wall and an opening extending around its main vessel axis as well as the use and a manufacturing method for such a reaction vessel.

Such, also referred to as sample vessels, reaction vessels are used, for example, in combinatorial chemistry and drug discovery and are usually elongated cylindrically shaped beakers open at the top. The best known form of a reaction vessel is the test tube with a rounded bottom. However, the reaction vessels commonly used in drug discovery today mostly have flat vessel bottoms, since it is easier to arrange samples on the flat vessel bottoms.

Usually, individual or even several reaction vessels are used in reaction blocks or bioreactors, as are known, for example, from WO 02/14539 A1. In these devices, the samples in the reaction vessels are synchronously defined reaction conditions, e.g. exposed to different temperatures with a certain gas supply. The temperature ranges, e.g. thermal blocks surrounding the reaction vessels can range from less than -80 ° C to more than +200 ° C, with the vessel bottoms usually being heated and the upper vessel areas being cooled. The samples in these devices are also e.g. mixed by agitators and, if necessary, can already be analyzed in the reactors by measuring systems.

For secure mounting, the reaction vessels are used in mounts or tight-fitting recesses or bores of the thermoblocks, the sample vessels being held in the vertical direction in that they either stand on their vessel bottom or are suspended from an edge that protrudes in the region of the vessel opening. [0005] To seal against the external atmosphere, the mostly upward-facing vessel openings of the reaction vessels are closed by lids, closures or plugs. So that there is the possibility of intervention or the supply of gases to the reaction vessels, for example, during operation in the reactors, openings, flaps or gastight slotted membranes are arranged in the covers through which, for example, feed lines, sensors, stirrer drives, etc. are guided or Substances can be introduced or removed. In the case of bioreactors in particular, the vessel openings are enclosed by gas lids, so that the samples or media in the vessel can be gassed or the vessel openings can be flushed with gas, such as an inert gas. This creates a closed system with a controlled atmosphere in the area of the vessel opening.

[0006] These known seals have already proven themselves very well in the past. However, against the background of the constantly increasing protection requirements, there is a need for further improvement in the sealing of the reaction vessels.

The invention is therefore based on the object of specifying an improved sealing of the reaction vessels.

This object is achieved with the reaction vessel according to claim 1, the reaction vessel according to claim 10 and the use of the reaction vessel according to claim 20 and the reaction vessel produced according to claim 21. Further preferred embodiments of the invention are specified in the subclaims.

The invention thus relates to a basically known reaction vessel, with a vessel wall and an opening extending around its main vessel axis. The vessel wall is understood to mean an enclosure which forms an arbitrarily shaped vessel and which, in the case of a cup, thus includes both the vessel side wall and the vessel bottom. The main axis of the vessel is the axis that runs through the center of gravity of the vessel opening and the center of gravity of the vessel. In the case of rotationally symmetrical vessels, this is the axis of symmetry which, in the case of an elongated cylindrical cup, extends through its center of gravity parallel to the longitudinal outside of the cylinder.

The opening itself can extend over the entire cross-sectional width or only parts of a vessel side. In the case of beaker-like reaction vessels, the opening extends over the entire cross-sectional width and therefore also has the diameter of the vessel. However, embodiments with a smaller opening are also conceivable, in which the cup-like reaction vessel has a perforated lid. The reaction vessel according to the invention differs from the known reaction vessels in that it has at least one spring element arranged on an outside of the vessel wall and acting in the direction of the main vessel axis, which is connected in one piece to the vessel wall.

The invention has the advantage that several vessels can be sealed tightly in a simple manner via a common lid. The contact pressure is approximately the same for all vessels. Tolerances are also compensated for.

If the reaction vessel thus obtained is now inserted into a holder, for example into a bore in a thermoblock, the reaction vessel is at least partially supported on the holder by the at least one spring element. Unlike previously, the reaction vessel is therefore not held or stored as rigidly as possible but deliberately movable. This has the advantage that the reaction vessel can still move in the holder at least in the direction of the main vessel axis. Due to the spring force of the spring element, the reaction vessel is therefore pressed away from the holder against the cover relative to a cover placed on the opening. This already considerably improves the sealing of the reaction vessel compared to a rigidly held reaction vessel.

If the recess of the thermoblock is slightly wider than the reaction vessel, the reaction vessel can also change in its angular position. So e.g. If the lid is pressed obliquely onto the edge of the vessel surrounding the vessel opening, the vessel will adapt to the angular position of the lid. This ensures that, even in the event of deviations in the angular positions of the two sealing sides to be brought into active connection with one another, that is to say the lid sealing surface and the vessel sealing surface, both surfaces always lie closely against one another and an optimal seal is thus achieved.

With a spring element extending and acting in the direction of the main vessel axis, such as e.g. a spiral spring, there is also the effect that the vessel does not slide completely into the recess, but rather is kept resiliently spaced from the bottom or the upper edge of the recess. The spring element, so to speak, presses the vessel away from the holder in the direction of its main vessel axis, so that the reaction vessel protrudes in the direction of its main vessel axis in comparison to an unsprung reaction vessel in relation to the recess or the holder. This makes it easier to remove the spring element.

Preferably, the spring elements are integrally connected, for example by injection molding, casting, welding or molding, in one piece with the vessel wall. This enables the spring elements to be fastened well to the outside of the vessel wall. It is irrelevant where the spring elements are arranged on the outside of the vessel wall. For example, the springs can sit under the bottom of the vessel or anywhere on the side wall of the vessel.

Preferably, the spring element is a resilient tongue projecting outward from the vessel wall. The simplest embodiment of such a spring element is therefore simply an elongated, resilient, rigid element attached to the wall of the vessel. This then develops a resilient effect due to its lower bending stiffness compared to the vessel wall. Such spring tongues can be attached to the vessel wall very simply and inexpensively.

In an alternative embodiment of the reaction vessel, the spring element is a bending strip held on two sides. It is therefore an elongated component, also referred to as a bending beam held on two sides, with a flat cross-sectional profile, the narrower cross-sectional side specifying the main bending direction, that is, the direction of action of the spring. Bending strips are also easy to manufacture, but have a better fastening than a spring tongue held on one side.

[0019] The spring element preferably has at least one pressure piece extending in the direction of the main vessel axis. The pressure piece specifies a support point for the spring element and can expediently be designed as an elevation on the spring element. This pressure piece point-like force into the ^'spring element then takes place. If the reaction vessel with the at least one spring on its outside is inserted into a recess for holding purposes, the reaction vessel is supported by the pressure piece on the spring element at the edge or on the bottom of the recess. Therefore, the pressure piece is preferably designed as a pin extending in the direction of the main vessel axis and connected in one piece to the spring element. The pressure piece thus serves to introduce force and to define a stroke by which the reaction vessel protrudes in the direction of the main vessel axis relative to the holder.

The bending strip preferably corresponds to the peripheral shape of the reaction vessel. So it can be guided around the outside of the vessel wall or be designed on the underside of a vessel bottom following this. In the case of a reaction vessel with a circular cross section, the bending strip is at least a section of a circular ring, for example a quarter circular ring. It is only important that the bending strip roughly follows the circumferential shape, for example as a circular ring section with a slightly elliptical cross section. Of course, angular circumferential shapes are also feasible, in particular polygonal shapes, the strip then being designed to be angled accordingly. It is often advantageous that the at least one spring element is not attached to the bottom of the vessel but to the side wall of the vessel wall. Therefore, the vessel wall preferably has an outwardly projecting edge on which the at least one spring element is arranged. In cases in which the reaction vessel is inserted vertically downwards into a recess for the holder, this projecting edge is expediently arranged on the uppermost side in the area of the vessel opening. However, if the reaction vessel is pushed vertically from below into an opening from below, it is also expedient to arrange this protruding edge at the bottom in the region of the bottom of the vessel. The protruding edge not only takes over the mounting of the spring element but also serves as a handle on which the vessel can be pulled out of a recess.

In order to keep the reaction vessel well, three spring elements are arranged distributed over the circumference of the vessel wall. These can be attached, for example, to the bottom of the vessel or to the side wall of the vessel or the protruding edge. The triple support of the reaction vessel results in a statically determined resilient mounting, which means that the reaction vessel does not tilt when the spring elements are sufficiently rigid.

Due to the spring elements, the reaction vessel can be pressed or pulled in any direction, however, essentially in its main vessel axis, without the reaction vessel tipping over. This means that with regard to its tilting stability, it does not matter how the reaction vessel is pressed or in which direction the reaction vessel is pressed, it will not fall over. If the three spring elements are attached to the protruding edge, it is even no longer necessary for the reaction vessel to abut a recess somewhere in the region of its side wall in order to be sufficiently stabilized. Thus, this statically determined resilient mounting leads to the greatest possible free space for the reaction vessel with regard to its angular position adjustment compared to a sealing element applied to the opening for sealing.

Preferably, the three spring elements are arranged in segments on the edge of the vessel wall and surrounded on their outer sides by a protective ring. This embodiment has the advantage that the spring elements themselves form a resilient edge and are well protected against damage. These spring segments are also particularly easy to produce. With an outwardly projecting annular edge on the vessel wall, the spring elements can be formed by slots running in the circumferential direction or by slots running radially in the direction of the main vessel axis. The spring segments are then either simply held radial segments or double held circumferential segments, which are surrounded on the outside by a protective ring to protect them from breaking off. This also has the advantage that, for example, injuries to the operating personnel can occur very sharp-edged spring elements can be excluded. The slots required for this can be formed particularly easily, for example in a casting mold, by appropriately shaped elevations in the edge area.

So that the reaction vessel can always be introduced into a reaction block or reactor in a defined orientation, positioning means for the positional alignment of the vessel are also arranged on the vessel wall, preferably on its protruding edge. The positioning means are expediently coding lugs and coding notches which are in engagement with corresponding counterparts, for example a holder, or adjacent reaction vessels. Thus, the reaction vessel introduced into a reaction block is always brought into an orientation that is the same with respect to the angular position by these positioning means. Several reaction vessels can also be easily aligned with one another via the positioning means. For this purpose, the positioning means of adjacent reaction vessels are brought into engagement and the reaction vessels against one another in their position, e.g. fixed with tape. It is also expedient to design the positioning means as a coupling so that the vessels can be chained to one another.

The object on which the invention is based is also achieved by a reaction vessel with an at least partially translucent vessel wall, in which the vessel wall has at least one optical lens in the translucent area. This lens is used for the optical evaluation of the reaction processes taking place in the reaction vessel and changes on the samples. Thus, fluorescent reaction substances can be attached to the vessel wall in the area of the lens and can be irradiated with light from outside through the lens for oxygen or CO ₂ measurement. Such analysis methods are described, for example, in WO 02/14539 A1.

In contrast to the prior art, part of the measuring device, namely the optics, is attached directly to the reaction vessel. If the reaction vessel is now closed in a gastight manner with a lid, it is no longer necessary to take the orientation of the vessel with respect to the optical measuring devices into account when closing the reaction vessel. Sealing is therefore given top priority, which improves it. In addition, the loading of the reaction vessels with sample material is simplified, since the lenses clearly show where the sample material, namely in front of the lens, has to be arranged. This significantly improves the measurement results. Finally, the height of the reactor is also reduced, since the distance between the lens and the sample material becomes smaller, while the light yield increases as a result of the lens being integrated into the vessel wall.

The lens is preferably arranged in a region of the vessel wall forming a vessel bottom. This is as flat as possible in order to simplify the arrangement of sample material. chen. The optical measuring devices such as light emitting diodes (LED) and the like can thus be arranged below the reaction vessel. On the one hand, this enables a complete lateral covering of the reaction vessel, for example with thermoblocks, and on the other hand, the arrangement of several vessels in rows and columns next to each other.

The lens is best connected in one piece to the vessel wall, for example cast onto the vessel wall. This results in a homogeneous and stable connection between the lens and the vessel wall, which means that no optical errors from reflections occur in the area of the attachment of the lens. Alternatively, the lens is glued to the vessel wall. For example, particularly high-quality lenses made of glass can be glued to a plastic tube wall.

The lens preferably has a convex lens contour which is suitable for the customary optical evaluation and measurement methods. The lens contour expediently extends on the outside of the vessel wall, as a result of which a smooth wall surface results in the interior of the vessel. Particularly when the lens is attached to the bottom of the vessel, it is easily possible because of the then smooth bottom of the vessel to align and fasten the sample materials on the bottom of the vessel exactly above the lens.

[0031] In a particularly preferred embodiment of the reaction vessel, two lenses are arranged on the wall of the vessel at a defined distance from one another. This makes it possible to arrange at least two different optical evaluation and measuring devices below or next to the reaction vessel. Measurements with different light spectra can also be carried out if, for example, a red light-emitting diode is arranged in the reaction vessel and a blue light-generating and UV-light-emitting diode is arranged below on the bottom of the vessel.

Preferably, the vessel wall is translucent and is light-scattering around the lenses. This serves to reduce stray light that, for example, penetrates from the outside or comes from a second adjacent measuring head. In an overall transparent embodiment of the vessel wall, the area around the lenses is expediently made matt, so that the light penetrating through this area of the vessel wall is refracted several times.

To improve the adhesion of reactants, the bottom of the vessel is pretreated on the inside at least in some areas with plasma. This guarantees good adhesion of the sample materials inside the reaction vessel, so that the samples are stirred with stirring bars without the reaction substances moving away from these areas. Particularly preferred is an embodiment in which the reaction vessel for lateral storage from the outside is acted upon by at least one outer wall segment which acts resiliently on the vessel wall. The resilient outer wall segment expediently forms, together with at least one rigid but preferably also resilient counterpart, a recess in which the reaction vessel is held on its side wall and the diameter of which is variable from a smaller than the outer diameter of the vessel to a corresponding one. Particularly advantageous is an embodiment with four outer wall segments which are resilient and heatable in the radial direction with respect to the main vessel axis and which center the reaction vessel in the recess. For the best possible heat conduction from the outer wall segments to the reaction vessel, the segments should rest directly on the reaction vessel and therefore have a shape that corresponds as closely as possible to the outer shape of the reaction vessel. The diameter of the recess is thus widened from a smaller diameter than that of the outer diameter of the vessel to a corresponding one when the reaction vessel is inserted into the recess.

According to the invention, such a reaction vessel is used in a bioreactor, the opening of the reaction vessel interacting with a cone, a spherical surface or another tapering, rotationally symmetrical surface of a gas hood. For this purpose, the reaction vessel is inserted into a holder such as a recess in a thermoblock of the bioreactor and then brought into its end position with the gas hood via a cone or the like of the gas hood.

For this purpose, the cone is, if possible, a body corresponding to the geometry of the vessel opening. If the vessel opening is circular, the cone is, if possible, a circular truncated cone. The cone then tapers away from the gas hood, from a diameter which is larger than that of the vessel opening to a diameter which is narrower than that of the vessel opening.

If the cone formed in this way is pressed into the vessel opening of the reaction vessel, the reaction vessel centers itself on the cone sliding into the vessel opening, since it can still move due to its resilient holder. The outer surface of the cone and the inner edge ring surface of the vessel opening then lie perfectly against one another and thus form a particularly tight seal. To further improve the seal, the inside edge of the vessel can then be rounded.

According to the invention, the reaction vessel is made in one piece as a cup from a transparent plastic by injection molding. Thus, both the vessel wall and the spring elements and the optical lenses are expediently also made in a single method step made in one piece from a single material. This greatly reduces the manufacturing costs and enables the inexpensive mass production of the reaction vessels as disposable products. A clear, transparent polystyrene plastic can be used as the transparent plastic.

The invention is explained in more detail below with reference to an embodiment shown in the drawing. It shows schematically:

Fig. 1 is a spatial view of the reaction vessel;

2 shows the section A-A through the opening area of the reaction vessel; and

Fig. 3 shows the section A-A when a cone is pressed into the opening of the reaction vessel.

The illustrated embodiment of the reaction vessel 1 is an elongated, cup-shaped, transparent injection molded part made of polystyrene. The reaction vessel 1 has a vessel wall 2 which comprises a circular vessel bottom 3, an externally circular-cylindrical vessel side wall 4 and an edge 5 which projects outwards. With the exception of the protruding edge 5, the reaction vessel 1 is basically formed rotationally symmetrical to the main vessel axis 6. The main vessel axis 6 therefore runs through the center of the vessel bottom 3 parallel to the vessel side wall 4 through the centroid of the vessel opening 7.

The flat vessel base 3 has on its outside two convex optical lenses 8 and 9 that are curved outwards. Both lenses 8 and 9 are arranged at the same radial distance from the main vessel axis. The lenses are used for an oxygen and / or CO ₂ measurement on fluorescent reaction substances which are attached to the vessel base 3 just above the lenses. Outside of the lenses 8, 9, the bottom of the vessel is transparent but, in contrast to the side wall 4 of the vessel, is roughened or matted in a refractive manner.

On the opposite side of the reaction vessel 1 is in the region of the vessel opening 7 the outwardly projecting edge 5, which in particular has three segment-like spring elements 10. In the exemplary embodiment shown here, the three spring elements of the same size are bending strips 11, which are held at their two ends in a manner that is resistant to bending, that is to say they transmit torque.

The three spring elements 10 are all the same size and correspond in shape to the circumferential shape of the reaction vessel, that is to say they are approximately three-circle-shaped. There are radially extending holding segments 12 between the spring elements. A bending strip 11 is thus held between two holding segments 12 of the protruding edge 5. A peg-shaped pressure piece 13 is formed on the underside of each bending strip 11 and in the middle of a single bending strip 11 arranged. Each pressure piece 13 extends parallel and in the direction of the main vessel axis 6 in order to introduce a bearing force acting in the direction of the main vessel axis from the weight of the sample and the weight plus any cone press-in force into the spring. On their outside, the three spring elements 10 are surrounded by a circumferential protective ring 14. This protects the spring elements 10 against unintentional breaking off, since the bending strips 11 are each thinner than the rest of the protruding edge 5. -Alternatively, the protective rings can be dispensed with and the spring elements can be guided up to the outer circumference of the edge.

The reaction vessel 1 is used in a bioreactor. For this purpose, it is inserted into a recess 16 for operation. In the exemplary embodiment shown in FIGS. 2 and 3, the recess is formed by outer wall segments 15 which surround the reaction vessel from the outside in a segment shape. These outer wall segments 15 are mounted in a radially resilient manner with respect to the main vessel axis 6 and press laterally from the outside onto the reaction vessel 1. In the embodiment shown here, however, the outer wall segments 15 are not yet applied to the vessel wall, but are held spaced apart from them in a prestressed manner.

The outwardly projecting vessel edge 5 has a larger outer diameter than the recess 16 and therefore projects laterally beyond it. The sample vessel now lies on the three pressure pieces 13 of the bending strips 11, which extend below the rim 5 of the vessel, and does not rest on them. the rim 5 itself. The weight of the reaction vessel 1 thus rests on the three pressure pieces 13, which each represent a pressure point for the bending strips 11. Due to this three-point bearing, it is a statically determined, but spring-loaded system.

If, as shown in FIG. 3, a gas hood 18 with a cone 19 tapering in the direction of the reaction vessel 1 is pressed onto the reaction vessel 1 from above, the circular conical cone 19 slips into the opening 7 of the reaction vessel 1 and lies on annular inner edge 20 of the opening 7 evenly. Due to the axially resilient mounting via the three bending strips 11 and the resilient lateral mounting via the outer wall segments 15, the reaction vessel 1 can still move resiliently or slightly twist relative to the cone 19.

This leads to a slanted reaction vessel 1 sliding along the cone outer surface and the vessel 1 changing its position until the outer surface of the cone 19 and the inner edge 20 of the vessel opening 7 lie perfectly against one another. The centering and sealing of the reaction vessel 1 thus takes place automatically when the gas hood 18 is arranged.

Finally, the protruding edge of the reaction vessel also has a coding lug 21 and a coding notch 22 as positioning means. With these positioning means, for example, eight Vessels 1 are chained to one another in a precisely aligned position with terminal strips or adhesive strips on the edge 5 and are handled very well as a bundle of vessels. The coding lug 21 also serves the coding notch 22 for positioning the vessel 1 in the bioreactor on corresponding counter-positioning means attached there.

Claims

1. reaction vessel (1) with a vessel wall (2) and an opening (7) extending around its main vessel axis (6), characterized in that there is at least one on an outside of the vessel wall (2) and in the direction of the main vessel axis (7 ) Acting spring element (10) which is integrally connected to the vessel wall (2).

2. Reaction vessel according to claim 1, characterized in that the spring element (10) is a resilient tongue projecting outwards from the vessel wall (2).

3. Reaction vessel according to claim 1, characterized in that the spring element (10) is a bending strip (11) held on two sides.

4. Reaction vessel according to one of the preceding claims, characterized in that the spring element (10) has at least one pressure piece (13) extending in the direction of the main vessel axis (6).

5. Reaction vessel according to one of the preceding claims, characterized in that the bending strip (11) corresponds to the peripheral shape of the reaction vessel (1).

6. Reaction vessel according to one of the preceding claims, characterized in that the vessel wall (2) has an outwardly projecting edge (5) on which the at least one spring element (10) is arranged.

7. Reaction vessel according to one of the preceding claims, characterized in that three spring elements (10) are arranged distributed over the circumference of the vessel wall (2).

8. Reaction vessel according to claim 7, characterized in that the three spring elements (10) are arranged in segments on the projecting edge (5) of the vessel wall (2) and are surrounded on their outside by a protective ring (14).

9. Reaction vessel according to one of the preceding claims, characterized in that positioning means (21, 22) for the positional alignment of the vessel are arranged on the vessel wall (2), preferably on its projecting edge (5).

10. reaction vessel (1) with an at least partially translucent vessel wall (2), characterized in that the vessel wall (2) has at least one optical lens (8; 9) in the translucent area.

11. Reaction vessel according to claim 10, characterized in that the lens (8; 9) is arranged in a region of the vessel wall (2) forming a vessel bottom (3).

12. Reaction vessel according to one of the preceding claims, characterized in that the lens (8; 9) is integrally connected to the vessel wall (2).

13. Reaction vessel according to one of claims 10 or 11, characterized in that the lens (8; 9) is glued to the vessel wall (2).

14. Reaction vessel according to one of the preceding claims, characterized in that the lens contour extends on the outside of the vessel wall (2).

15. Reaction vessel according to one of the preceding claims, characterized in that the lens (8; 9) has a convex lens contour.

16. Reaction vessel according to one of the preceding claims, characterized in that two lenses (8, 9) on the vessel wall (2) are arranged at a defined distance from one another.

17. Reaction vessel according to one of the preceding claims, characterized in that the vessel wall (2) is designed to be translucent and light-scattering around the lenses (8, 9).

18. Reaction vessel according to one of the preceding claims, characterized in that the bottom of the vessel (3) is pretreated on the inside at least in regions with plasma for improved adhesion of reactants

19. Reaction vessel according to one of the preceding claims, characterized in that the reaction vessel (1) for lateral storage from the outside is acted upon by at least one outer wall segment (15) acting resiliently on the vessel wall (2).

20. Use of a reaction vessel (1) according to one of the preceding claims in a bioreactor, wherein the opening (7) of the reaction vessel (1) cooperates with a cone (19) of a gas hood (18).

21. The production method for a reaction vessel (1) according to one of the preceding claims, characterized in that the reaction vessel (1) is produced in one piece as a cup using the injection molding process.

22. The method according to claim 21, characterized in that the reaction vessel (1) is made of transparent plastic.