Centrifuge bottle for countercurrent operation

Abstract

A centrifuge bottle (1) for countercurrent operation, comprising a bottle mouthpiece (2) arranged radially outward with respect to a centrifugal rotation axis, and a bottle end (3) opposite the bottle mouthpiece (2) arranged radially inward with respect to the centrifugal rotation axis, and comprising a sample outlet, a tube (8) for connecting the bottle mouthpiece (2) to a sample inlet, a cavity (12) with an outer circumference that varies over the cross-section of the centrifuge bottle (1) from the bottle mouthpiece (2) to the bottle end (3), wherein the outer circumference of the cavity (12) is determined by at least three angles α, β and γ extending from a central axis of the centrifuge bottle (1), and at least one bottle end region (7) adjoining y, wherein β defines a mouthpiece section (5) of the centrifuge bottle (1) extending from the bottle mouthpiece (2) to a Intermediate section (4) extends,wherein α defines the intermediate section (4) extending between the mouthpiece section (5) and a rear section (6), wherein γ defines the rear section (6) extending between the intermediate section (4) and the bottle end region (7), characterized in that the value of γ is greater than the value of α.

Description

[0001] The invention relates to a centrifuge bottle for countercurrent operation according to the preamble of claim 1.

[0002] Centrifuge bottles are used for countercurrent operation in centrifuges. Countercurrent operation of centrifuges serves to concentrate and separate a specific fraction of a sample within the centrifuge bottle(s) of a centrifuge rotor. Separation occurs through the suspension of cells or particles in a fluidized bed by applying a constant flow force against the centrifugal forces. Due to their different sizes and densities, various cells or particles exhibit different sedimentation properties, which are exploited for their separation in the countercurrent process.

[0003] The centrifuge bottles typically have a conical shape with a substantially cylindrical section towards the rotor center, as shown, for example, in WO 2022 / 046572 A1, where a centrifuge bottle with a conical mouthpiece section and a subsequent cylindrical section is disclosed.

[0004] Therefore, an equilibrium between the settling velocity and the flow velocity of the sample within these centrifuge bottles can only be achieved at a specific radius of the centrifuge bottle. This typically leads to a concentration of the sample particles at this specific radius, accompanied by particle loss or particle pelleting, both of which are undesirable as they can distort the analysis of a specific cell or particle content of the sample.

[0005] The objective of the present invention is to improve the sample flow within the centrifuge bottle, so that improved particle separation is achieved through countercurrent operation.

[0006] This problem is solved by the subject matter of the independent patent claim.

[0007] Advantageous embodiments and further developments are the subject of the dependent claims, the associated description and the figures.

[0008] According to the invention, the problem is solved by a countercurrent centrifuge bottle comprising a bottle mouthpiece arranged radially outwards with respect to a centrifugal rotation axis, wherein a bottle end is arranged radially inwards relative to the bottle mouthpiece with respect to the centrifugal rotation axis and comprises a sample outlet, a tube for connecting the bottle mouthpiece to a sample inlet, and a cavity with an outer circumference that varies across the cross-section of the centrifuge bottle from the bottle mouthpiece to the bottle end. In a preferred embodiment, the radius of the sample outlet is larger than the radius of the sample inlet.

[0009] The centrifuge bottle is suitable for countercurrent operation in centrifuges. The bottle is therefore designed to be rotated about a central axis, and the shape of the cavity, defined by the three angles, allows the forces acting on the sample to be balanced. When separating cells or particles from a sample, it is important to note that smaller particles / cells settle more slowly than larger ones; and to optimize the volume of the centrifuge bottle, the shape of the cavity should be adjusted accordingly, which is made possible by the centrifuge bottle according to the present invention.

[0010] The outer circumference of the cavity is determined by at least three angles α, β, and γ, originating from a central axis of the centrifuge bottle and at least one bottle end region connected to γ. Angle β defines a nozzle section of the centrifuge bottle extending from the nozzle to an intermediate section. The cross-section of the nozzle section must be chosen such that the fluid velocity is significantly higher than the settling velocity. This is advantageous to prevent pelleting of particles / cells. Angle α defines the intermediate section extending between the nozzle section and a rear section, and angle γ defines the rear section extending between the intermediate section and the bottle end region.The cross-section of the intermediate section can be selected based on the particle / cell density, fluid density, and viscosity of the individual sample being handled. Preferably, the desired centrifuge speed and sample flow rate are maximized, maintaining an equilibrium between the settling velocity caused by centrifugal force and the flow velocity. The intermediate section is the portion of the cavity where this equilibrium can be achieved. For accurate calculations, it is crucial that the value of γ is greater than the value of α. The rear section acts as a barrier for sample particles. It is designed such that the settling velocity is higher than the fluid velocity. This prevents particles from escaping the bottle, which could lead to inaccurate sample analysis.The angle γ is preferably located in a range where a factor of at least 2 is maintained between the settling velocity and the flow velocity of the sample. This is advantageous for increasing the separation efficiency.

[0011] The inlet and outlet sections of the centrifuge bottle are preferably thinner than the other sections to prevent sedimentation of sample particles. A further advantage of the present invention is the improved separation performance of the centrifuge bottle and the prevention of pelleting. The centrifuge bottle is designed with a rear section acting as a barrier for improved separation.

[0012] In a preferred embodiment, the mouthpiece is rounded. A rounded mouthpiece advantageously directs the sample flow from the tube into the centrifuge bottle. The angles can be adjusted depending on the type of sample being centrifuged. They can be calculated to match a specific particle density, fluid density, or mass flow rate of the centrifugation process in order to achieve improved results.

[0013] The shape of the cavity of the centrifuge bottle allows for a reversal of the countercurrent flow of the sample when it is introduced into the bottle using the tube.

[0014] According to one embodiment of the invention, the value of β is smaller than that of α. For example, β can be between approximately 0° and approximately 45°. A preferred range is between 4° and 16°, and particularly preferably between 4° and 8°. The larger angle α results in a larger volume flowing towards the center of the bottle, and a smaller angle β towards the mouthpiece results in an increased sample fluid velocity and a more uniform flow. This combination contributes to improving the overall separation efficiency of the centrifuge bottle. To concentrate a specific particle / cell size of a sample, the bottle size and shape can be individually adjusted to achieve the desired effect by modifying one or more of the angles α, β, and γ.

[0015] According to a further embodiment of the invention, the value of β can be positive, negative, or 0°. The β value can be individually adjusted so that the bottle can be tailored to specific sample properties. For example, particle density, fluid density, mass flow rate, and particle size of the samples to be separated can be included in the calculation of a preferred angle β.

[0016] According to another embodiment, the intermediate section is longer than the mouthpiece section and longer than the rear section. The volume of the intermediate section should be the largest compared to the other sections in order to establish the force balance in this section for improved particle / cell concentration.

[0017] According to another embodiment, the bottle end region has a constant circumference. Preferably, the radius of the bottle end region corresponds to the greatest extent of the preceding rear section. This allows the conditions of the rear section to be maintained for a longer period. This is advantageous for sample flow and also simplifies the manufacture of the centrifuge bottle. This part can be kept short compared to other sections of the bottle, particularly compared to the intermediate section.

[0018] According to a further embodiment, the bottle end region comprises a first region with a constant circumference and a second region with a constant circumference. The diameters of the first and second regions can be different to create a stepped bottle end region. The diameters of the regions can be selected to suit the samples to be analyzed and their composition, for example, with regard to density and / or particle structure. This can advantageously further improve the accuracy of the analysis.

[0019] According to another embodiment, the diameter of the first constant-circumference section is larger than the diameter of the second constant-circumference section. This reduces the weight and volume of the centrifuge bottle, and consequently, the manufacturing costs. A reduced volume due to a smaller bottle end section also reduces the forces exerted on the centrifuge bottle by the centrifugal force during countercurrent operation. This has a positive effect on preventing damage to the centrifuge bottle that could render the sample unusable and ensures the proper functioning of the centrifuge bottle.

[0020] However, it would also be possible to provide a section with a decreasing perimeter instead of a section with a constant perimeter following the section defined by γ.

[0021] According to another embodiment, the tube is a central tube extending within the cavity. Depending on the intended use and the type of sample, this type of tube routing in the center of the cavity may be most suitable for achieving improved results.

[0022] According to another embodiment, the bottle mouthpiece is a doubly rounded mouthpiece that includes a radially inwardly oriented tip for eccentrically diverting the sample flow. The bottle mouthpiece can, for example, be designed as a B-shaped mouthpiece to ensure a uniform, two-part sample flow from the tube running inside the cavity and from the mouthpiece into the cavity of the centrifuge bottle. The combination of the central tube projecting into the cavity with the doubly rounded mouthpiece, which includes a radially inwardly oriented tip, can improve the sample flow, since the sample flowing through the tube located centrally in the cavity of the centrifuge bottle can then be diverted equally into the cavity, thus allowing for more precise application.

[0023] According to another embodiment, the tube is arranged outside the cavity. The tube can, for example, run along the outer surface of the centrifuge bottle. This arrangement can be advantageous for saving space within the cavity of the centrifuge bottle. EXAMPLE

[0024] In an exemplary embodiment of the invention, a centrifuge bottle for countercurrent operation is provided with the following dimensions: Mouthpiece radius 0,015 m Mouthpiece distance a 0,0006 m Angle β 67,00° angle α 4,50° angle γ 25,00° Distance from the center to the mouthpiece 0,20 m Length of angle α 0,085 m Length of γ 0,055 m Length of the constant area 0,015 m Radius of the first constant region 0,050 m Radius of the second constant area 0,020 m

[0025] This type of centrifuge bottle is designed for optimized use under the following conditions: Input speed 3000 rpm Volume flow rate V 0,000 m 3 / s Tube diameter D 0,006 m Kinematic viscosity 1,01 E-06 m 2 / s density Flüssigkeit 1.000 kg / m 3 density Partikel 1.100 kg / m 3 Particle diameter 1.00 E-05 m

[0026] This example can be modified depending on the type of sample to be centrifuged. The better the centrifuge bottle is adapted to a specific sample type and the environmental parameters, the more precisely the sample can be fractionated.

[0027] Further features, details and advantages of the invention will become apparent from the wording of the claims and from the exemplary embodiment described below with reference to the drawings.

[0028] The accompanying drawings, which are included in this patent specification and form an integral part thereof, illustrate embodiments of the invention and, together with the above general description and the following detailed description of the embodiments, serve to explain the principles of the present invention.

[0029] The following applies to the drawings: Fig. 1 is a side view of a centrifuge bottle for countercurrent operation according to an embodiment of the invention; Fig. Figure 2 is a side view of a countercurrent centrifuge bottle with a double rounded mouthpiece; and Fig.Figure 3 is a side view of a centrifuge bottle for countercurrent operation with an external inlet.

[0030] Fig.Figure 1 shows a centrifuge bottle 1 according to embodiments of the present invention. The centrifuge bottle 1 comprises a bottle mouthpiece 2 and a bottle end 3 opposite it. Extending from the bottle mouthpiece 2 between the bottle mouthpiece 2 and the bottle end 3 are a mouthpiece section 5, an intermediate section 4, a rear section 6, and subsequently a bottle end region 7. In this embodiment, the bottle end region 7 is divided into a first region with a constant circumference 10 and a second region with a constant circumference 11. It would also be possible to provide the bottle end region 7 with only a constant circumference. Furthermore, it would also be possible to provide a bottle end region 7 that is divided into a region with a constant circumference and a region whose circumference decreases towards the bottle end 3.The precise design can be adapted to the individual properties of different samples, ensuring consistently high measurement accuracy.

[0031] The outer dimensions of the mouthpiece section 5 are defined by an angle β, those of the intermediate section 4 by an angle α, and those of the rear section by an angle γ, each measured from the central axis A. The angle γ is significantly steeper than α and β. The circumference of each section 4, 5, and 6 increases towards the end of the bottle 3.

[0032] Centrifuge bottle 1 can be used in a centrifuge to separate samples into different components, for example, to separate cells from a sample based on their size and density, which influence their sedimentation behavior. Cells with lower density and / or smaller diameter can be separated from the sample and eluted. This ensures proper separation of sample fragments for further analysis and can be used, for example, to separate blood cells. Three different angles α, β, γ define the outer circumference of the bottle from the mouthpiece to the end of the bottle.

[0033] The bottle mouthpiece 2 is preferably a rounded mouthpiece, as shown in this embodiment. The angle β of the mouthpiece section 5 of the portion of the centrifuge bottle 1 following the bottle mouthpiece 2 is steeper than that defining the bottle mouthpiece 2. This serves to reverse the sample flow within the centrifuge bottle 1. The angle α of the intermediate section 4 is selected to ensure a balance between flow rate and settling rate. This intermediate section 4 defines the portion of the centrifuge bottle 1 in which a fluidized bed is achieved, which is crucial for the proper separation of particles from the sample to be analyzed. Following the intermediate section 4, the rear section 6 is shown. The angle γ, which determines the course of the outer circumference of the rear section 6 from the intermediate section 4 towards the bottle end region 7, is significantly steeper than α.This creates a barrier for the particles of the sample to be analyzed. The angle γ should be within a range that maintains a settling velocity at least twice the flow velocity. The bottle end region can be divided into different sections. As shown in this embodiment, it is possible to provide a first section with a constant circumference 11. The dimensions of these sections can be defined by the manufacturing process of the centrifuge bottle 1. The diameter over the entire first section 10 corresponds to the largest extent of the rear section 6. In this embodiment, the bottle end region 7 has a second section with a constant circumference 11 following the first section 10.The constant circumference of the second region 11 has a significantly smaller diameter than the first region 10 in order to reduce weight and volume, thereby minimizing the stress exerted on the centrifuge bottle 1 and lowering manufacturing costs. However, it is also possible to design the bottle end region 7 differently, for example, as a single-piece region with a constant circumference, or as a single-piece region with a decreasing or increasing circumference, depending on the desired embodiment and the substances to be analyzed, or with a first region 10 with a constant circumference and a second region 11 with a constant circumference that do not differ significantly from each other, but with the circumference of the second region 11 being only slightly smaller than that of the first region 10. The [reference to be added] Fig.In the embodiment shown in 1, the space is used for particle concentration in the cavity 12, so that good separation can be achieved by the rear section 6 acting as a barrier section.

[0034] The mouthpiece 2, which in this embodiment is a rounded mouthpiece, is advantageous for preventing pelleting of particles / cells of the sample to be centrifuged. The shape of the centrifuge bottle 1 achieves a high particle / cell concentration in the intermediate section 4 and highly efficient separation in the rear section 6. Overall, the centrifuge bottle 1 exhibits improved separation performance when used in countercurrent operation.

[0035] Fig. 2 and Fig. Figure 3 represents embodiments of centrifuge bottles 1 with different variants of bottle mouthpieces 2. Apart from the variants of bottle mouthpieces 2, the following correspond to those shown in Figure 3: Fig. 2 and Fig.3 depicted centrifuge bottles 1 in their design, in particular the external shape, of the centrifuge bottle 1 in Fig. 1.

[0036] Fig. Figure 2 shows a centrifuge bottle 1 with a double-rounded mouthpiece 9. The double-rounded mouthpiece 9 has a B-shape at its tip, which, after exiting the tube 8 through its double-rounded mouthpiece 9, ensures a continuous flow in two separate halves due to the radially inward-oriented tip. The tube 8 is a central tube that extends along the central axis within the cavity. In this way, the flow is not interrupted but is nevertheless evenly separated and distributed into the cavity 12, which can be advantageous for certain types of samples. Fig.Figure 3 shows an embodiment of the centrifuge bottle 1 for countercurrent operation with a modified bottle mouthpiece 2 connected to a tube located outside the cavity 12 of the centrifuge bottle 1. This allows the sample to be fed into the centrifuge bottle 1 while simultaneously saving space inside the centrifuge bottle 1. Depending on the type of sample being processed, different mouthpiece and / or tube types can be selected to ensure optimal performance. Reference sign 1 centrifuge bottle 2 bottle mouthpieces 3 bottle ends 4 Intermediate section 5 Mouthpiece section 6 rear section 7 Bottle end area 8 tubes 9 double rounded mouthpiece 10 first area with constant perimeter 11 second area with constant perimeter 12 Cavity α Angle α β angle β γ angle γ QUOTES INCLUDED IN THE DESCRIPTION

[0000] This list of documents cited by the applicant was automatically generated and is included solely for the reader's convenience. The list is not part of the German patent or utility model application. The DPMA accepts no liability for any errors or omissions. Cited patent literature

[0000] WO 2022 / 046572 A1

[0003]

Claims

[1] Centrifuge bottle (1) for countercurrent operation, comprising: a bottle mouthpiece (2) which is arranged radially outwards with respect to a centrifugal rotation axis; wherein one end of the bottle (3) is arranged radially inwards relative to the bottle mouthpiece (2) with respect to the centrifugal rotation axis and includes a sample outlet; a tube (8) for connecting the bottle mouthpiece (2) to a sample inlet; a cavity (12) with an outer circumference that changes across the cross-section of the centrifuge bottle (1) from the bottle mouthpiece (2) to the bottle end (3); wherein the outer circumference of the cavity (12) is determined by at least three angles α, β and y extending from a central axis of the centrifuge bottle (1) and at least one bottle end region (7) adjoining y, where β defines a mouthpiece section (5) of the centrifuge bottle (1) extending from the bottle mouthpiece (2) to an intermediate section (4), where α defines the intermediate section (4) which extends between the mouthpiece section (5) and a rear section (6), where γ defines the rear section (6) which extends between the intermediate section (4) and the bottle end region (7), characterized by , that the value of γ is greater than the value of α. [2] Centrifuge bottle (1) according to claim 1, characterized by , that the value of β is smaller than that of α. [3] Centrifuge bottle (1) according to claim 1 or 2, characterized by , that the value of β can be positive, negative, or 0°. [4] Centrifuge bottle (1) according to any one of claims 1 to 3, characterized by , that the intermediate section (4) is longer than the mouthpiece section (5) and longer than the rear section (6). [5] Centrifuge bottle (1) according to any one of claims 1 to 4, characterized by , that the bottle end region (7) has a constant circumference. [6] Centrifuge bottle (1) according to any one of claims 1 to 4, characterized by , that the bottle end region (7) comprises a first region with constant circumference (10) and a second region with constant circumference (11). [7] Centrifuge bottle (1) according to claim 6, characterized by , that the diameter of the first area with constant circumference (10) is larger than the diameter of the second area with constant circumference (11). [8] Centrifuge bottle (1) according to any one of claims 1 to 7, characterized by , that the tube (8) is a middle tube running inside the cavity (12). [9] Centrifuge bottle (1) according to claim 8, characterized by, that the bottle mouthpiece (2) is a double rounded mouthpiece (9) with a radially inwardly oriented tip for eccentric deflection of the sample flow. [10] Centrifuge bottle (1) according to any one of claims 1 to 7, characterized by , that the tube (8) is arranged outside the cavity (12).

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