EP2334435A2

EP2334435A2 - Modular system of functional units for mixing, processing and/or separating samples for use in biological/medical research and for diagnostics

Info

Publication number: EP2334435A2
Application number: EP09806070A
Authority: EP
Inventors: Jan-Michael Heinrich; Christoph Mohr
Original assignee: Universitaet Leipzig
Current assignee: Universitaet Leipzig
Priority date: 2008-10-06
Filing date: 2009-10-06
Publication date: 2011-06-22
Also published as: WO2010040347A4; WO2010040347A2; DE112009003386A5; US20110287472A1; WO2010040347A3; DE102008050750A1

Abstract

Basic steps of biological research and of diagnostics include mixing, physical and/or chemical reactions and separation. Particularly in research task-specific laboratory systems are required which are not commercially available due to small batch sizes and the specific problems to be faced. Because of the lack of such systems, researchers and diagnosticians have to improvise with already existing vessels, filters, centrifuges etc. to solve the problems. The aim of the invention is to devise a system of functional units which can be put together depending on the methodological type of problem. The system of functional units which can be freely combined consists of mixing cylinders (2), separation devices (3) and a flow-regulating connecting unit (5) which is preferably a vacuum unit and which is designed to be preferably interconnected in any order with any number of functional units via simple plug-in connections to give a self-contained sample processing system.

Description

Modular system of functional units for mixing, processing and / or separating samples for use in biological / medical research and diagnostics

description

Mixing, physical and / or chemical reactions and separation are fundamental steps in biological research and diagnostics. In particular, research requires task-specific laboratory systems that are not commercially available because of small series sizes and the specific problem. In the absence of these systems, it is necessary to improvise with existing vessels, filters, centrifuges, etc. to solve the problem.

In practice, it is also necessary for smaller series of samples to be processed to perform several successive steps by transferring, pipetting, filtering, centrifuging, etc. of the samples, the samples must be successively transferred manually between different vessels, with a high workload requires a high material cost, since usually only a part of the steps is performed in each vessel. In addition to the factor "human", the different laboratory equipment of different laboratories (especially of vessels, pipettes, filters, etc.) often leads to different results being obtained in the individual laboratories with the same work instructions, so that elaborate optimization work for the reproducibility and / or or standardization of results in individual laboratories.

The aim of the invention is therefore to provide a system of functional units, which can be combined depending on the methodological task, preferably mated, and which allow a simple and reproducible mixing, physical and / or chemical reactions and separation of sample media.

This object is achieved according to the claims. The invention, which is realized according to the claims, therefore relates to a system according to claim 1 and its use according to claim 19. The further claims 2-18 and 20 are preferred embodiments of the invention.

Surprisingly, a modular system (hereinafter also referred to as "system" or "system according to the invention") consisting of one or more mixing cylinders, one or more separation devices and a flow-regulating connection unit, which are combined with one another in an application-specific manner for the most diverse requirements, has proven to be particularly simple and simple reproducible system for processing sample media.

The invention accordingly relates to a system of freely combinable functional units which is suitable in particular for mixing, processing and / or separating samples

- at least one mixing cylinder (2)

- At least one separating device (3) and

- A flow-regulating connection unit (5), wherein the flow-regulating connection unit (5) is preferably formed as a vacuum unit, wherein the system is designed so that the functional units (2; 3; 5), preferably via simple connectors, in any order , In particular with any number of functional units, are connected to a preferably self-contained, sample processing system.

The functional units according to the invention therefore preferably consist of or are

- mixing cylinder (s)

- Separator (s) and

- a vacuum unit.

The system allows the use of different cylindrical (closed on one side) containers, such as sample tubes. This is done by scaling the Dimensions achieved. The cylinder-like containers can be used as a collection or waste container for the supernatants or the respective fractions.

An important criterion is the design of the transitions between the preferably assembled functional units. This connection is preferably achieved by a wedge-shaped geometry. In order to prevent sample material from drawing into the sealing space between the functional units by capillary forces, at least one seal, in particular a line seal and preferably propylene, is used. As sealing possibility different variants come into question (see for example illustration 3G, 3H). Variant one is straightened in the lower part of the wedge, so that a circumferential connection between two functional units comes to conditions. Advantageously, variant one has an easier to produce by injection molding geometry. Variant two is a circumferential ring at the end of the plug-in connection, which is preferably attached directly to the spline connection by injection molding. This leads to an additional ring connection. In addition, it is possible to produce this seal as an O-ring or by 2K injection molding. An inclined plane on the top of the separation units (backwash level) allows a directed backwashing of the retentate from the filter surface while reducing the amount of backwash liquid. The backwashing level according to the invention is preferably an arrangement of several, preferably 6-18, webs which are arranged on the outside of the separating unit (3) or protrude from the separating unit (3), the webs being formed in this way or arranged so that the underside of the webs at one in relation to the top and / or bottom of the separation unit (imaginary) oblique (essay) plane ends or through the bottom of the webs one, compared to the top or bottom of the Separation unit (3), (imaginary) inclined (essay) level when placing the separation unit (3) is formed on a collecting container (7). The slope of the (imaginary) plane is preferably formed over an angle of 2-45 °, preferably 5-40 °.

Preferably, spacers (32) prevent the separating device (3) during the backwashing with the wall of the collecting container (7), which is preferably formed as a tube, comes into contact. The individual components or functional units of the modular system according to the invention will be explained in more detail below.

1st mixing cylinder

The mixing cylinder is preferably open on both sides, but can also be open only on one side (so that one would tip the sample material on the separating device), and is on the one side by a lid which preferably has a living hinge, or as a screw cap, plug or pluggable Cover executed on the lid, and on the other side closed by a peelable and / or pierceable foil and / or a pluggable, pluggable or screw-on bottom cap.

The mixing cylinder is used for sample collection and can be used as an alternative to single-sided open sample tubes. The interior is designed so that it can serve as a mixing vessel for achieving biological, chemical or physical reactions. The geometry of the mixing cylinder ensures

low shear forces, especially when incubating with biological material such as primary cells, cell lines, etc.

- minimal dead spaces

- Maximum sample recovery, and is suitable for preferably for incubation on a roll mixer as well as for

Incubation on an overhead mixer.

Preferred features for meeting the requirements are:

Both sides of the mixing cylinder are designed with different geometry: - filling side (but can also be used to dump the liquid on the separating device):

The filling side is closed by a lid with a film hinge (for other closure options see above). From here, the filling of the mixing cylinder with sample material and optional reaction material and solutions. The film hinge allows aseptic working. For the use of the mixing cylinder in combination with mechanical mixing devices, the lid has a peripheral edge. The peripheral edge avoids snagging or sticking of the cylinder during the mixing process (in particular on a tilting roller mixer roller mixer) to an unobstructed rotation of the To ensure mixing cylinder with film hinge, preferably the lid is additionally modified with a peripheral edge. - Discharge side (can also be used as a filling side):

The discharge side of the mixing cylinder is designed so that it can be plugged into the separation device, stuffed or screwed with this. The drainage side can optionally be closed with a foil and / or bottom cap (see above: preferably by inserting plugs or screws). The foil (one-time use) is attached to the cylinder by heat (thermowelded). If necessary (e.g., after incubation with fabrics), the film is removed by peeling off through the attached protrusion or by a sharp object such as e.g. a pipette tips, or pierced by a puncture device directly to the severing device / perforated. The use of a bottom cap allows aseptic work. The entire fitting area / insertion area of the mixing cylinder is closed by the bottom cap. By using the bottom cap, the discharge side of the mixing cylinder can be used simultaneously as the filling side. The discharge side can optionally be equipped directly with a sieve, which allows, for example, the pre-separation of desired or the deposition of undesirable fractions. Retained desired fractions can be used directly or reused by resealing the mixing cylinder with the bottom cap for further biological, chemical or physical reactions. The design of the cylinder, when combined with a sieve on the discharge side, allows it to be combined directly with the flow regulating connection unit, and thus can be used similarly to the separation device. Retained unwanted fractions increase the purity of the subsequent separation with the separation device.

Both sides of the mixing cylinder are preferably the same: each side can be used as a filling and / or emptying side, wherein the filling and / or emptying side with a lid, in particular with one of the above-mentioned lid, eg with a losierbaren cover, are formed , The direction for combination with a separating device is freely selectable. Each of the sides can be connected to the severing device. 2. Separator

The separation devices are based on the principle of filtration. The filter material and the pore size are provided according to the task. It can serve both separation by size separation, as well as the immobilization of substances. As the separators themselves serve as vessels, they may also serve as a reaction space for chemical reactions (e.g., activation and / or coupling of proteins) as well as ingest substances having selective properties (e.g., with particular affinities).

3. Flow regulating connection unit (adapter), which is preferably formed as a vacuum unit.

Vacuum unit in the sense of the invention means in particular a transition piece with (laterally arranged) intake, preferably for the connection with a vacuum pump, so that by connecting the vacuum unit (5) to the separation vessel (7), eg Screwing / clamping the vacuum unit (5) on or with the separation vessel (7), a suction bottle is made.

The flow-regulating connection unit allows the connection of the separation device with different cylindrical (closed on one side) containers, such as sample tubes and serves as an adapter. The shape is correspondingly adapted on one side to the geometry of the separating device and on the opposite side to the geometry of the respective cylinder-like containers. The connection unit is responsible for a defined liquid flow and supports the separation unit in the application of at least one, preferably more than two, separation device (s). By closing the connection unit, liquid can be kept in the separation device for incubation purposes (eg for immunochemical reactions). After opening the closure of the connection unit, the liquid flows into the cylinder-like container. Depending on the task, the target substance can either be in the run, immobilized on the membrane or attached to solid supports which have been introduced into the reaction space of the separation device. The connection unit can be connected to the collection and / or waste vessel either via a thread or via a wedge construction.

Experience has shown that depending on the manufacturer, the tubes differ greatly in the increase of the thread, but the inner diameter of the tubes only varies by a few micrometers. This can preferably be compensated by a simple wedge construction of the vacuum unit. The fixation preferably takes place as a "press fit." This results in a system which offers the user the opportunity to use his existing tubes.

The connection unit allows the combination with a vacuum filtration by connection of vacuum generating devices and equipment combinations (among other things disposable syringes, peristaltic pump, vacuum pump, water jet pump, pipettor with hose etc. Vacuum pressure, vacuum generating devices) The following is the one shown in Figure 4E and 4F , Principle of vacuum filtration briefly explained and serves as an example for the use of vacuum generating devices and device combinations.

The vacuum unit was designed so that a liquid movement takes place exclusively via the movement of the disposable syringe (57). Since the system is closed, no liquid from the mixing cylinder (2) on the separation unit (3) can run into the collecting vessel (7). The membrane material used serves as an additional barrier and prevents the compensation of air from the collecting vessel against the liquid in the receiving vessel. When the syringe is drawn up, a negative pressure forms in the collecting vessel, which conveys the liquid from the collecting vessel via the separating membrane into the collecting vessel. The two check valves (56) allow the air to be pushed out of the spray without the air being forced back into the separation units.

Further advantageous features and embodiments of the system according to the invention are also apparent from the figures, which reflect various embodiments of the modular system and its functional units. Figure 1A shows a 3D representation of a system according to the invention with a mixing cylinder (2) with film hinge lid, two separation units or

Separating devices (3), a flow-regulating connection unit (5), which is formed as a vacuum unit (5), and a collecting vessel (7), which are inserted into one another.

Figure 1 B shows a side view of the system shown in Figure 1A.

Figure 1C shows a side sectional view of the system according to Figure 1A

Figure 1D shows a view of the system according to Fig. 1A from above.

Figure 1 E shows a view of the system as shown in Fig. 1A from below.

Figure 2A shows a side view of the mixing cylinder (2) with open

Cover.

Figure 2B shows a side view of the mixing cylinder (2) with the lid closed.

Figure 2C shows a rear view of the mixing cylinder (2) with closed

Cover.

Figure 2D shows a sectional view of the mixing cylinder (2) with the lid open

Figure 2E shows a sectional view of the mixing cylinder (2) with closed

Cover.

Figure 2F shows a view of the mixing cylinder (2) with the lid open from below.

Figure 2G shows a view of the mixing cylinder (2) with the lid open from the top.

Figure 2H shows a view of the mixing cylinder (2) with the lid open from below.

Figure 2I shows a view of the mixing cylinder (2) with the lid open from below.

Figure 3A shows a 3D representation of the separation device (3).

Figure 3B shows a side view of the separating device (3).

Figure 3C shows a front view of the separator (3).

Figure 3D shows a sectional view of the separating device (3).

Figure 3E shows a top view of the separator (3).

Figure 3F shows a view of the separator (3) from below.

Figure 3G: shows a line seal variant of the separator (3).

Figure 3H shows another line seal variant of the separator (3). Figure 31 shows a composite of two separation units (3), wherein preferably the upper of the two separation units (3) has a filter or a separation membrane with a pore diameter x and the lower of the two separation units (3) has a filter or a separation membrane with a Pore diameter <x so that a sieve cascade is made.

Figure 3K shows a sectional view of the two separation units (3) with variants of the line seal.

Figure 3L shows an illustration of a separation unit (3) with a backwash level, which allows directional backflushing.

Figure 3M shows a side view of a separation unit (3) (with sieve) with

Backwashing device for backwashing the retentate after use of the system according to the invention, and after the separation unit (3) of the other

Functional units of the modular system has been separated and with a further receptacle (6), e.g. a centrifuge tube (6) has been connected.

Figure 3N shows a side view of the arrangement according to Fig. 3M.

Figure 30 shows a sectional view of the arrangement according to Fig. 3M.

Figure 4A shows a 3D representation of the vacuum unit (5).

Figure 4B shows a side view of the vacuum unit (5).

Figure 4C shows a sectional view of the vacuum unit (5).

Figure 4D shows a top view of the vacuum unit (5).

Figure 4E shows a system according to the invention with connection of a

Vacuum device - before filtration.

Figure 4F shows a system according to the invention with connection of a

Vacuum device - after filtration.

Figure 4G shows a structure of a system according to the invention for detaching cells on the sieve surface of the separation unit (3).

Figure 4H shows a sectional view of a structure for detaching cells from the membrane surface of the separation unit (3), in particular the construction according to FIG

Fig. 4G.

FIG. 5A shows a schematic representation of a closure mixing cylinder (2) with a bottom cap (8) or a mixing cylinder (2) which can be closed with a bottom cap (8). Figure 5B shows a sectional view of the bottom cap fixation. Figure 5C shows a sectional view of required geometry on the mixing cylinder for fixing the bottom cap.

The inventive system of freely combinable functional units consisting of

at least one, preferably one, mixing cylinder (s) (2)

- At least one, preferably two to eight, separating device (s) (3) and

- A flow-regulating connection unit, which is preferably formed as a vacuum unit (5) is designed so that the functional units, preferably via connectors, in any order, in particular with any number of functional units, are connected to a, preferably self-contained, sample processing system, in particular for mixing, editing and / or separating samples.

In this case, the system is preferably formed, in particular in the dimensions of its functional units, so that it can be brought onto or be connected to commercially available containers, in particular laboratory tubes, preferably with a volume of 5-100 ml, preferably a filling volume of 15 ml or 50 ml can.

In a preferred embodiment, the system according to the invention is characterized in that the mixing cylinder (2) can be temporarily closed on both sides by both sides.

In another preferred embodiment of the system, the interior of the mixing cylinder or cylinders is designed in such a way that optimum mixing of the samples is made possible.

In a further advantageous embodiment of the invention, the system is designed so that the separation device (s) on the principle of filtration consists or exist, ie in particular that the separation device (s) (each) a filter, a membrane and / or a sieve have or have or are configured with it. The filters, membranes and / or screens preferably have pores with a pore diameter of 10 nm-1 mm. In yet another preferred embodiment, the at least one separating device is designed so that membranes can be used for separating the material, which have a pore diameter between 10 nm and 1 mm.

In a particularly preferred embodiment of the system according to the invention, the membrane (s) consist of known material (s) which can be used both as size filters and / or as solid carriers with immobilizing surface design ,

In another particularly preferred embodiment, the system according to the invention is characterized in that the at least one separation device can be used as a reaction vessel for chemical and / or physical reactions.

A further advantageous embodiment relates to a system according to the invention, whose at least one separating device can be provided with openings for charging or emptying if required.

Another advantageous embodiment relates to a system according to the invention, in which a flow-regulating connection unit with three-way cock forms the end of the sample processing system.

In a particularly advantageous development of the system according to the invention, residence time and / or flow time of the sample into or through the functional units can be freely regulated by the flow-regulating connection unit.

Yet another advantageous embodiment of the system according to the invention is characterized in that it can be connected to commercially available vessels for receiving the flow.

Particularly preferred is a system according to the invention that

a mixing cylinder (2)

- Two to eight, preferably two to five, particularly preferably two or three, separating devices (3) and - A flow-regulating connection unit, in particular a

Vacuum unit (5), which are arranged in series in this sequence, wherein preferably the two to eight separating devices (3) are formed by suitable selection of the pore size as a sieve cascade, i. the filter, membrane or sieve pore diameter of the mixing device from top to bottom, i. decreases in the direction of the flow-regulating connection unit, so that, for example, microparticles of different sizes can be separated via this cascade sieve.

In particular, an inventive system is preferred, whose at least one mixing cylinder (2) at least one of the features 20-29 and / or 80-86 or preferably a combination of such features or more preferably all features 20-29 and / or 80-86 according to the list of reference numerals and / or the illustrations.

Another preferred system according to the invention is characterized in that the at least one separating device (3) comprises at least one of the features 30-42 or preferably a combination of such features or more preferably all features 30-42 according to the list of reference numerals and / or the drawings.

Yet another preferred system according to the invention is characterized in that the flow-regulating connection unit (5), which is preferably formed as a vacuum unit (5), at least one of the features 50-60 or preferably a combination of such features or more preferably all features 50-60 according to the list of reference numerals and / or the figures has.

Another preferred embodiment relates to a system according to the invention, in particular suitable as a separating device, which further comprises (as a further functional unit) a collecting and / or collecting container (7), which is preferably connected to the flow-regulating connecting unit (preferably vacuum unit) (5). Another advantageous embodiment of the invention relates to a, in particular designed as a separation unit, system, which is designed according to Figures 1A-1 E.

Another aspect of the invention relates to the use of the system according to the invention for use in biological and / or medical research, for diagnostics, and / or for product control in biotechnology, in particular for mixing, processing and / or separating samples.

Further advantageous properties and features of the invention will become apparent from the following non-exhaustive embodiments.

Application Example 1

Isolation of specific cells from a blood sample - exemplified by the separation of CD3 - positive cells from whole blood followed by

(A) Isolation of RNA or DNA

(B) Recovery of single cells

(C) backwashing the particle-cell complexes

(D) Use of the specifically depleted sample

explained.

Depending on the application, a mixing cylinder, a separation device, a flow-regulating connection unit and two collection containers are required. Whole blood supplemented with anticoagulants is transferred to the mixing cylinder with anti-CD3 particles on the filling side. The mixing cylinder is closed and rotated on a roll mixer for at least 10 and a maximum of 45 minutes. Following incubation, the mixing cylinder is opened on the discharge side. On the cylinder, the separation device is then spent with a collecting container. The construct of mixing cylinder, separating device and collecting vessel is rotated. Subsequently, the mixing cylinder is opened at the filling side. The mixture of whole blood and anti-CD3 particles then passes over the separator. The particles, which have captured the CD3-positive cells from the whole blood, remain on the membrane of the separator. Unbound and unwanted cells are rinsed by repeated rinsing with a washing solution into the collecting vessel.

(A) The separation device together with the particle-cell complexes is then placed on a fresh, combined with a flow-regulating connection unit, sample tube. The connection unit is closed by a cap so that no exchange of air and liquid can take place through the separation device. Subsequently, a lysis liquid for the release of RNA or DNA from the cells is placed in the reaction space of the separation device and incubated. After incubation, the cap of the connection unit is removed and replaced with a disposable syringe. The entire liquid, including the RNA from the CD3-positive cells, is then transferred into the collecting vessel with the aid of the syringe. The particles remain on the membrane. The purification of the RNA or DNA from the cell lysate is then carried out using the methods of separation of RNA or DNA according to the state of the art.

(B) The separation device together with the particle-cell complexes is then placed on a fresh, combined with a flow-regulating connection unit, sample tube. The connection unit is closed by a cap so that no exchange of air and liquid can take place through the separation device. Subsequently, a special separation liquid for separating the cells from the particles is placed in the reaction space of the separation device and incubated. After incubation, the cap of the connection unit is removed. The cells are transferred to the collecting vessel by repeated rinsing with a washing solution. The specific cells of CD3-positive cells, free from unwanted cells, are now available to the user for further analysis.

(C) The separation device together with the particle-cell complexes is rotated and placed on a fresh sample tube. Rinsing with a wash solution rinses the particle-cell complexes into the fresh tube. These Fraction can then be used, for example, for culturing the cells directly on the particle.

(D) The unwanted cells, which were rinsed by repeated rinsing with a washing solution in the collecting vessel can be used as a depleted fraction for further analysis.

Application Example 2:

Cascade application for simultaneous isolation of different specific

Fractions from a blood sample - exemplified by the separation of CD4- and CD8-positive cells from whole blood, as a rapid test for the determination of HIV-immune status, explained.

Application related are a mixing cylinder, two separator, two

Flow regulating connection unit and at least three

Collecting containers needed.

Depending on the application, two different particle sizes are needed:

- small particles that catch the CD4-positive cells

- medium particles that capture the CD8-positive cells

According to the two particle sizes, two separators with different pore sizes are needed:

- small pore size to support the small particles

- Large pore size to support the large particles

Whole blood supplemented with anticoagulants is transferred to the mixing cylinder with the small anti-CD4 particles and the large anti-CD8 particles on the filling side. The mixing cylinder is closed and rotated on a roll mixer for at least 10 and a maximum of 45 minutes. Following incubation, the mixing cylinder is opened on the discharge side. On the cylinder then the separation device with the large pore size and then the separation device is spent with small pore size. Finally, the collection container is positioned on the last separation device. , The construct of mixing cylinder, separating devices and collecting container is then rotated and opened the mixing cylinder on the filling side. The large CD8 particles together with the cells remain on the membrane of the first separation device, the smaller CD4 particles pass the first separation device and remain on the lower separation device, unwanted as well as unbound cells happen the separation devices and are collected in the collecting vessel. Each of the separation devices together with the particle-cell complexes can then be used as a separate fraction. To determine the HIV-immune status, a ratio of the isolated CD4 to CD8 cells is formed. The points (A) - (D) of the application example one can also be applied to each of the fractions.

Application Example 3

Cascade application for simultaneous isolation of different cytokines for the specific determination of inflammatory status from a blood sample. Depending on the application, a mixing cylinder, three separation device, three flow-regulating connection unit and at least four collection containers are required. According to the application, three different particle sizes are needed:

- small particles catching IL-6

- medium particles that capture IL-8

- large particles that catch TNFalpha

According to the two particle sizes, three separation devices with different pore sizes are needed:

- small pore size to support the small particles

- average pore size for the retention of the middle particles

- Large pore size to support the large particles

The isolation of the specific cytokines is carried out as described in application examples one and two. Each of the separated fractions is then incubated with chemicals on the appropriate separation unit, so that the loading of the particles with the respective protein can be detected by a color change. The intensity of the color change in the reaction mixture can be determined with the aid of a simple measuring device. The measured values obtained can then be used to generate a corresponding diagnosis with regard to the inflammation status. Legend to the pictures:

Figure 1A: 3D illustration of 2 separation units Figure 1 B: Side representation of separation units Figure 1C: Side sectional view of separation units Figure 1 D: Top separation view Figure 1E: View of separation units from below

Figure 2A: Side view of the mixing cylinder with lid open Figure 2B: Lateral view of mixing cylinder with lid closed Figure 2C: Rear view of mixing cylinder with lid closed Figure 2D: View of the mixing cylinder with lid open Figure 2E: View of the mixing cylinder with lid closed Figure 2F Figure 2G: View of the mixing cylinder with lid open from the bottom Figure 2H: View of the mixing cylinder with open lid from the bottom Figure 2I: View of the mixing cylinder with the lid open from the bottom

Figure 3A: 3D illustration of a separation device or separation unit

Figure 3B: Side view of a separating device

Figure 3C: Front view of a separator

Figure 3D: Section view of a separator

Figure 3E: Top View of a Separator

Figure 3F: View of a separator from below

Figure 3G: line seal variant

Figure 3H: Line seal variant

Figure 3I: Composite of 2 separators

Figure 3K: Sectional view of the 2 separation devices with variants of the

line seal

Figure 3L: Illustration of the backwash level

Figure 3M: Side view of a screen for backwashing the retentate

Figure 3N: Side view of a screen for backwashing the retentate

Figure 30: Sectional view of the structure for backwashing the retentate Figure 4A: 3D representation of the vacuum unit Figure 4B: Side view of the vacuum unit Figure 4C: Sectional view of the vacuum unit Figure 4D: Top view of the vacuum unit

Figure 4E: Connecting a vacuum device - before filtration Figure 4F: Connecting a vacuum device - after filtration Figure 4G: Structure for detaching cells on the sieve surface Figure 4H: Sectional view of the structure for detaching targets from the membrane surface

Figure 5A: Schematic representation of the closure of the mixing cylinder with a

bottom cap

Figure 5B: Sectional view of the bottom cap fixation Figure 5C: Sectional view of the required geometry on the mixing cylinder for the

Fixation of the bottom cap

LIST OF REFERENCE NUMBERS

1 . .. separation units

2. .. mixing cylinder

3. .. separation membrane

5. .. vacuum unit

7. .. collection / collection container

8th . .. bottom cap

20 ... movie hinge

21 ... peripheral edge

22 ... Opening for opening the lid

23 ... cover

24 ... outer wall of the mixing cylinder

25 ... surface fit

26 ... line seal

27 ... Inner wall of the mixing cylinder

28 ... sealing film

29 ... tear-off tongue for the sealing foil 30 ... Outer wall of a separation unit above the handle

31 ... handle

32 ... spacers

33 ... bridges

34 ... Outer wall of a separation unit under the handle

35 ... surface fit

36 ... line seal

37 ... recess for ventilation on the outer wall of the separation unit under the handle

38 ... recess for ventilation on the handle

39 ... separation membrane

40 ... backwashing

41 ... backwash reservoir

42 ... drainage edge

50 ... outer wall of the vacuum purity

51 ... male connection

52 ... Surface fit vacuum of the vacuum unit

53 ... line seal

54 ... Noses of the connecting piece

55 ... output solution

56 ... check valve

57 ... disposable syringe 58 ... filtrate

59 ... Relief reservoir

60 ... gap to prevent liquid suction with the

vacuum device

80 ... External geometry of the bottom cap

81 ... Interior geometry of the bottom cap

82 ... clamping area 1

83 ... clamping area 2

84 ... clamping area 3 85 ... spacers

86 ... geometry of the mixing cylinder for fixing the bottom cap

All the features illustrated in the foregoing description, the appended claims and the drawings may be relevant to the realization of the invention in its various forms both individually and in any combination.

Claims

claims

1. System of freely combinable functional units consisting of

- at least one mixing cylinder (2)

- At least one separating device (3) and

- A flow-regulating connection unit, which is preferably formed as a vacuum unit (5) designed so that the functional units, preferably via connectors, in any order, in particular with any number of functional units, are connected to a, preferably self-contained, sample processing system, in particular for Mixing, editing and / or separating samples.

2. System according to claim 1, characterized in that it can be spent on commercial containers.

3. System according to any one of claims 1 or 2, characterized in that the mixing cylinder (2) can be temporarily closed on both sides of both sides.

4. System according to any one of claims 1-3, characterized in that the interior of the mixing cylinder is designed so that an optimal mixing of the samples is made possible.

5. System according to any one of claims 1-4, characterized in that the separating devices are based on the principle of filtration.

6. System according to any one of claims 1-5, characterized in that membranes can be used for separating the material with a pore diameter between 10nm and 1 mm.

7. System according to any one of claims 1-6, characterized in that the membranes consist of known materials and can be used both as a size filter as well as a solid support with immobilisierender surface design.

8. System according to any one of claims 1-7, characterized in that the separation devices can serve as a reaction vessel for chemical and / or physical reactions.

9. System according to any one of claims 1-8, characterized in that the separating devices can be provided if necessary with openings for loading or unloading.

10. System according to any one of claims 1-8, characterized in that a flow-regulating connection unit with three-way valve forms the conclusion of the sample processing system.

11.System according to claim 9, characterized in that by the

Flow regulating connection unit residence time and flow time of the sample in or through the functional units can be freely regulated.

12. System according to any one of claims 1-11, characterized in that it can be connected to commercially available vessels for receiving the flow.

13. System according to any one of claims 1-12, characterized in that it is made

a mixing cylinder (2)

- One to five, preferably one, two or three, separating devices (3) and

- A flow-regulating connection unit (5), which is preferably formed as a vacuum unit (5) consists, which are arranged in series in this sequence.

14. System according to any one of claims 1-13, characterized in that the at least one mixing cylinder (2) has the features 20-29 and / or 80-86 according to the list of reference numerals and / or the figures.

15. System according to any one of claims 1-14, characterized in that the at least one separating device (3) has the features 30-42 according to the list of reference numerals and / or the figures.

16. System according to any one of claims 1-15, characterized in that the flow-regulating connection unit (5), which is preferably formed as a vacuum unit, the features 50-60 according to the list of reference numerals and / or the figures.

17. System according to any one of claims 1-16, in particular designed as a separation unit, characterized in that it further comprises a collecting and / or collecting container (7), preferably with the flow-regulating connection unit (5), which is preferably formed as a vacuum unit , connected is.

18. System according to claim 17, in particular designed as a separation unit, characterized in that it is designed according to Figures 1A-1 E.

19. Use of the system according to any one of claims 1-18 for use in biological and medical research, for diagnostics and for product control in biotechnology.

20. Use according to claim 19 for mixing, processing and / or separating samples.