Sample cartridge for incubating and / or analyzing dispersions of particles, cells, or droplets.

Abstract

To provide a sample cartridge for incubating and / or analyzing a dispersion of particles, cells or droplets.SOLUTION: The present invention relates to a sample cartridge for incubating and / or analyzing a dispersion of particles, cells or droplets and / or for performing biochemical reactions with or in such dispersion. The present invention furthermore relates to a device for incubating a dispersion of particles, cells or droplets and / or for performing a biochemical reaction therewith. Moreover, the present invention also relates to use of a sample cartridge or of a device for generating and / or processing a dispersion of particles, cells or droplets. Moreover, the present invention relates to a method of processing a dispersion of particles, cells or droplets. Furthermore, the present invention relates to a method of generating a dispersion of droplets and to a method of generating a dispersion of solid or semi-solid particles.SELECTED DRAWING: Figure 4

Description

[Technical Field]

[0001] The present invention relates to a sample cartridge for incubating and / or analyzing dispersions of particles, cells, or droplets, and / or for carrying out biochemical reactions with or in such dispersions. The present invention further relates to a device for incubating and / or carrying out biochemical reactions with dispersions of particles, cells, or droplets. Furthermore, the present invention also relates to the use of a sample cartridge for generating and / or processing dispersions of particles, cells, or droplets. Furthermore, the present invention relates to a method for processing dispersions of particles, cells, or droplets. [Background technology]

[0002] Many biological and chemical test procedures or processes require incubation of particle suspensions or droplet emulsions at one or more defined temperatures and over one or more defined time periods. To ensure comparable results, the particles or droplets must be exposed to one or more precisely defined temperatures. This can be achieved by continuous or sequential movement of the sample, or by reducing the cross-section of the reaction space. A flat reaction chamber in close contact with the temperature-controlling and defining element allows for rapid heat transfer, which is particularly useful in situations of temperature cycling or rapid heating or cooling steps that may be required by certain assay processes. However, such flat chambers are assembled from different parts and typically sealed at the edges. This results in complex assembly procedures, high costs, and assembly artifacts that can have adverse effects on the performance of such assembled reaction chambers / cuvettes. One possible adverse effect concerns the formation of gas bubbles at the edges of the reaction chamber during the filling of such a reaction chamber. Gas bubbles can lead to temperature inhomogeneity, which can negatively affect the optical observation of any processes occurring within the reaction chamber. Another possible adverse effect relates to the relative rigidity design of the chamber, which can result in suboptimal heat exchange between the heater / cooler and the chamber. This is mainly due to small voids remaining between the surfaces. Therefore, it is desirable to have a chamber geometry that reduces the risk of such artifacts, further provides optimal heat transfer and temperature homogeneity, and enables optical observation of the reaction space.

[0003] In recent technologies, different devices have been described to facilitate the incubation of liquids. Traditionally, the simplest method involves using standard microreaction vials, which are allowed to be regulated to a defined temperature over long time periods. Many different types of microfabricated chambers have been used to improve mixing efficiency and temperature homogeneity. For example, EP 0 891 811 A1 describes a method and apparatus for mixing thin films of fluid. The device employs a mixing mechanism that induces mixing within a fluid chamber formed by two opposing surfaces, resulting in the agitation of the liquid within the fluid chamber. A similar setup with a microchip with an immobilized probe is described in FR 2803225. WO 03 / 015923 describes a microfluidic device with means for enabling fluid movement within a low-volume, low-aspect-ratio microfluidic chamber. All of the aforementioned devices represent micro-machined microsystems with microchannels constructed from different parts assembled to have at least two different surfaces. WO 2007 / 051861 discloses a device and method for particle detection, comprising a reaction chamber formed within a chamber body between a first surface and a second surface located opposite it. The device further includes one or more displacements that provide labeling and liquid displacement within the chamber. Digital techniques based on droplet emulsions have become an important approach in biological analysis, and therefore new tools and methods are required to facilitate the processing of emulsified samples. Such techniques are typically similar to the processing of cells or beads, and therefore improved solutions would benefit the processing of all types of analytes. For example, this application provides the following items. (Item 1) A sample cartridge (100) for incubating and / or analyzing a dispersion (200) of particles, cells, or droplets, in particular a suspension (210) of particles or cells, or an emulsion (220) of droplets, and / or for carrying out a biochemical reaction with or in such a dispersion, wherein the cartridge is A deformable permeable tube (110), Two opposing open ends (111, 112), the two opposing open ends each serving as an inlet and an outlet. A deformable permeable tube (110) having Equipped with, The tube is adapted to receive a dispersion of particles, cells, or droplets, particularly a suspension of particles or cells, or an emulsion of droplets, and within the internal space (113) of the tube, the internal space is lined by one wall (114'') or several walls (114, 114', 114'') of the tube, and the tube is configured such that when the internal space of the tube receives the suspension, emulsion, or dispersion, it has a circular or oval cross-section, and when the internal space is pressed against a surface, it has a flat, non-circular cross-section. The cartridge further comprises means (120) for retaining the particles, cells, or droplets within the deformable permeable tube, wherein the means for retaining the particles are located at one or both ends of the tube, and the means for retaining the particles are preferably a filter, membrane, grid, mesh, sieve, or other structure that retains the particles while allowing the passage of a liquid through it. (Item 2) The sample cartridge according to item 1, wherein the deformable permeable tube has a single wall (114''), and the internal space (113) is lined by the single wall. (Item 3) The sample cartridge according to any one of items 1-2, further comprising means (130) for reversibly closing and sealing the deformable permeable tube at one or both of the opposing ends (111, 112), preferably the means for reversibly sealing being a clamp, or, in the case of sealing at both ends, a pair of clamps located at the opposing ends. (Item 4) A sample cartridge according to any one of items 1-3, further comprising a mounting frame (140) connected to the tube at the opposing ends of the tube, holding it and configured to allow the addition of a substance, such as a liquid or solid or a mixture thereof, to the internal space of the tube via one of the ends which serves as an inlet, and / or the removal of a substance, such as a liquid or solid or a mixture thereof, from the internal space of the tube via one of the ends which serves as an outlet. (Item 5) The sample cartridge according to item 4, wherein the mounting frame (140) has first and second lateral sides (141, 142) positioned opposite to each other, one of the lateral sides is preferably formed of a permeable planar substrate configured to act as an opposing surface (321) against which the tube can be pressed, and the mounting frame is configured such that the other of the lateral sides of the mounting frame allows physical contact of the temperature control device to the central portion of the tube through the other of the opposing lateral sides, and allows the temperature control device to apply pressure to the central portion of the tube through the other of the opposing lateral sides, preferably against the opposing surface of the permeable planar substrate, thereby allowing exposure of the central portion of the tube (115) to the temperature control device. (Item 6) The sample cartridge according to any one of items 4-5, wherein the mounting frame (140) is further preferably configured to allow analysis of the central portion (115) of the tube by optical detection means through one of the lateral sides of the mounting frame, more preferably through the lateral side formed by the transparent planar substrate as defined in item 5. (Item 7) The sample cartridge according to any one of items 5-6, wherein the central portion (115) of the tube is the portion that is closed and sealed by the means (130) for reversibly closing and sealing the deformable permeable tube. (Item 8) The mounting frame has a longitudinal axis (143) aligned with the longitudinal axis (116) of the tube, and the mounting frame comprises two opposing longitudinal ends (144, 145), each of which has an orifice (146, 147) that fluidly connects to the opposing ends (111, 112) of the tube, each of which serves as the inlet and outlet of the tube, and each of the orifices is sealable and preferably comprises means (148, 148') for reversibly closing and sealing such orifice, such as a cap, tap, plug, stopper, or screw cap, as described in any of items 4-7. (Item 9) The sample cartridge according to item 8, wherein the tube (110) is mounted within the mounting frame (140) such that the opposing open end of the tube is attached to or in contact with the opposing longitudinal end of the mounting frame, and the mounting frame encompasses a space (149) through which the tube extends, and such space is configured to allow exposure to or contact of the central portion (115) of the tube to a temperature control device and / or to allow analysis of the central portion of the tube by optical detection means. (Item 10) The walls (114, 114', 114'') of the tube, preferably the single wall (114'''), have a thickness in the range of 1 μm to 1,000 μm, preferably 20 μm to 200 μm, more preferably 50 μm to 150 μm, and / or the diameter of the internal space is 0 when the internal space has a circular or oval cross-section.The height of the internal space of the tube is in the range of 1 cm to 5 cm, and / or when the tube has a flat, non-circular cross-section, it is in the range of 5 μm to 500 μm, preferably 5 μm to 200 μm, more preferably 10 μm to 150 μm, and preferably the deformable permeable tube is permeable in the range of 250 nm to 950 nm, preferably in the range of 400 nm to 600 nm, and / or in the range of 450 nm to 650 nm, and / or in the range of 500 nm to 700 nm, and / or in the range of 550 nm to 750 nm. The material is transparent in the range of 600 nm to 800 nm and, if present, allows for analysis of any contents in the internal space of the tube using optical spectroscopy and / or imaging, preferably the material is styrene-butadiene rubber, silicone rubber, polyvinyl butyral, polyurethane, polyisobutylene, polyhydroxybutyrate, polyhydroxyalkanoate, polyether block amide, rubber, gum arabic, isoprene rubber, fluororubber, ethylene vinyl acetate copolymer, ethylene Sample cartridges as described in any of the above items, selected from any combination or copolymer of the following: propylene-diene-rubber copolymer, ethylene-ethyl acrylate copolymer, chloroprene rubber, ethyl rubber, butadiene rubber, acrylonitrile-methyl methacrylate copolymer, acrylonitrile-chlorinate-polyethylene-styrene copolymer, acrylonitrile-butadiene-acrylate copolymer, polyester (PES), polyethylene terephthalate (PET), polyethylene (PE), high-density polyethylene (HDPE), polyvinyl chloride (PVC), polyvinylidene chloride (PVDC), low-density polyethylene (LDPE), polypropylene (PP), polystyrene (PS), high-impact polystyrene (HIPS), polyamide (PA), acrylonitrile-butadiene styrene (ABS), polyethylene / acrylonitrile-butadiene styrene (PE / ABS), polycarbonate (PC), polycarbonate / acrylonitrile-butadiene styrene (PC / ABS), polyurethane (PU), and any combination or copolymer of the above. (Item 11) Use of any of the sample cartridges described in items 1-10 for generating and / or processing dispersions of particles, cells, or droplets, in particular suspensions of particles or cells, or emulsions of droplets. (Item 12) The use described in item 11, wherein the processing is one or more of the following activities: incubating the dispersion of particles, cells, or droplets; carrying out a biochemical reaction with the dispersion of particles, cells, or droplets; binding one or more analytes to the particles, cells, or droplets and then removing any unbound analytes and other unbound substances from the particles, cells, or droplets; exchanging the liquid phase of the dispersion; and analyzing the dispersion of particles, cells, or droplets. (Item 13) The aforementioned use is, The steps include filling the tube with a dispersion of particles, cells, or droplets, particularly a suspension of particles or cells, and optionally one or more additional reagents; The steps include optionally arranging the particles, cells, or droplets in a single layer, The steps include carrying out a biochemical reaction in the tube and / or incubating the tube under one or more defined reaction conditions, in particular under one or more temperature conditions. The steps include analyzing the results of such biochemical reactions and / or such incubations, Uses described in any of items 11-12, including those listed below. (Item 14) The aforementioned use is, A step of filling a tube with a dispersion of particles, cells, or droplets, wherein the dispersion comprises a first liquid, preferably a first aqueous liquid. The steps include removing the first liquid from the particles, cells, or droplets in the tube by, for example, centrifugal force, gravity, or suction, while the particles, cells, or droplets are trapped by the trapping means, by allowing the first liquid to pass through the trapping means, for example, The steps include adding a second liquid containing the analyte, preferably a second aqueous liquid, to the tube, A step of incubating the particles, cells, or droplets in the second liquid to enable or facilitate the binding of the analyte to the particles, cells, or droplets, The steps include: washing the particles, cells, or droplets to optionally remove any unbound analytes and unbound substances from the particles, cells, or droplets, by, for example, centrifugation, gravity, or suction, by allowing the second liquid to pass through the means for containing the particles, cells, or droplets while the particles, cells, or droplets are contained by the means for containing the particles, cells, or droplets; The steps include: resuspending the particles in a third liquid, preferably a non-aqueous liquid, by adding such a third liquid to the tube; The steps include optionally arranging the particles, cells, or droplets in a single layer, The steps include carrying out a biochemical reaction in the tube and / or incubating the tube under one or more defined reaction conditions, in particular under one or more temperature conditions. The steps include analyzing the results of such biochemical reactions and / or such incubations, Uses described in any of items 11-13, including those listed below. (Item 15) A method for producing a droplet dispersion, in particular a droplet emulsion, the method comprising the steps of: providing a cartridge according to any of items 1-10; mixing an aqueous phase and an oily liquid phase within the tube of the cartridge; and thus producing a dispersion, in particular a droplet emulsion. (Item 16) A method for producing a dispersion of a solid or semi-solid, such as gel particles, the method comprising the steps of providing a cartridge according to any one of items 1 - 10, mixing an aqueous phase and an oily liquid phase within the tube of the cartridge, and thus producing a dispersion, particularly an emulsion of droplets, wherein one of the phases further contains solid particles or any one of one or several components capable of forming a gel or a solid in response to changing at least one environmental condition around the component, and when one of the phases further contains one or several components capable of forming a gel or a solid, the method further comprises changing the at least one environmental condition around the component, thereby converting the droplets into particles and inducing the formation of a gel or a solid by producing a dispersion of particles within the tube, preferably, the at least one environmental condition is selected from temperature, pH, pressure, light in a defined wavelength range, ultrasonic waves, and the presence of polymerization-inducing chemicals, the method. (Item 17) A device (300) for incubating a dispersion of particles, cells, or droplets, particularly a suspension of particles or cells, or an emulsion of droplets, and / or for performing a biochemical reaction therewith, the device comprising a sample cartridge (100) according to any one of items 1 - 10, a temperature control unit (310) having a temperature control surface (311) and adapted to heat and / or cool via the temperature control surface, and comprising the device is configured such that the tube of the sample cartridge, particularly the central portion (115) of the tube, can be brought into contact with or is in contact with the temperature control surface (311) using one of the lateral sides (141, 142) and can be pressed or is pressed against the temperature control surface (311), whereby the tube is deformed when pressed against or by the temperature control surface, and the internal space (113) of the pressed tube has a flat non-circular cross-section, the device. (Item 18) The device further comprises a facing unit (320), which is positioned at a distance opposite to the temperature control unit (310) and has a facing surface (321) facing the temperature control surface (311). When present within the cartridge, the facing unit forms one of the lateral sides (141, 142) of the cartridge and is configured to act as a facing surface (321) against which the tube can be pressed. The facing unit is either the permeable planar substrate or the facing unit (320) does not form part of the cartridge and is a separate component provided within the device (300) separately from the cartridge. The separate component is preferably configured to be operable to apply pressure via the facing surface (321) on the tube in contact with the temperature control surface (311), or is operable to be positioned at a defined distance with respect to the sample cartridge (109). The device according to item 17. (Item 19) One or both of the temperature control surface or the facing surface is preferably permeable in the range of 250 nm to 950 nm, preferably in the range of 400 nm to 600 nm, and / or in the range of 450 nm to 650 nm, and / or in the range of 500 nm to 700 nm, and / or in the range of 550 nm to 750 nm, and / or in the range of 600 nm to 800 nm. The device according to item 18. (Item 20) The device further comprises optical detection means (340), which are configured to be able to detect and / or analyze the contents of the internal space (113) of the tube using optical spectroscopy and / or imaging. Preferably, such detection and / or analysis is carried out using a beam path that passes through the central part (115) of the tube and one or both of the temperature control surface (311) or the facing surface (321). The device according to any one of items 17 - 19. (Item 21) Use of any of the devices described in items 17-20 for generating and / or processing dispersions of particles, cells, or droplets, in particular suspensions of particles or cells, or emulsions of droplets, accompanied by a sample cartridge described in any of items 1-10, and carried out as defined in any of items 11-14. [Prior art documents] [Patent Documents]

[0004] [Patent Document 1] International Publication No. 03 / 015923 [Patent Document 2] International Publication No. 2007 / 051861 [Overview of the Initiative] [Means for solving the problem]

[0005] In this field, there is a need for a reaction chamber that reduces the risk of artifact formation, such as the uncontrollable formation of gas bubbles, and enables optimal heat transfer, temperature homogeneity, and optical observation of the sample inside the reaction chamber. [Brief explanation of the drawing]

[0006] The present invention will be further explained herein by reference to the figures.

[0007] [Figure 1] Figure 1 shows a cross-section of a deformable permeable tube of a sample cartridge according to the present invention, containing a dispersion of particles, cells, or droplets. The cross-section is circular, and it can be seen that the tube is not pressed or in contact with any surface. [Figure 2] Figure 2 shows such a tube in contact with the temperature control unit and the opposing unit prior to being pressed. [Figure 3] Figure 3 shows the same tube being pressed between the temperature control unit and the opposing unit. [Figure 4]Figure 4 shows an embodiment of a sample cartridge according to the present invention. On the upper panel (a), there is a top view of an embodiment of the sample cartridge showing a mounting frame with two opposing longitudinal ends, each having an orifice closed by a screw cap. Also shown is one of the lateral sides formed by a permeable planar substrate configured to act as an opposing surface against which the tube can be pressed. The tube is theoretically visible through the permeable planar substrate but is not shown. Panels b)-d) show a cross-section along the longitudinal axis of panel a), and in addition, show means for reversibly closing and sealing the deformable elastic permeable tube in the form of a clamp, located at one end of the permeable tube, and means for retaining the tube itself and particles within the tube (shown as two dashed lines pointing to the right of the cartridge inside). In panel b), the means for reversibly closing and sealing is open, and in panels c)-d), it is closed, thus closing and sealing one end of the tube, and as a result, in panel c), the tube and its internal space adopt a wedge shape. In panel d), a temperature control device having a temperature control surface is pressed against the tube, thus pressing the tube against the opposing surface formed by the permeable planar substrate of the sample cartridge. Depending on the pressure applied and the dimensions of the tube, the internal space of the tube may have a height that is just sufficient to accommodate a single layer of particles, cells, or droplets. This allows for the analysis of individual particles without overlap. Furthermore, the cross-sectional areas shown in panels b)-d) are the two lateral sides of the mounting frame, which are located opposite each other, one of which is formed by a permeable planar substrate configured to act as an opposing surface against which the tube can be pressed, and the other of which is a mounting frame, which allows physical contact of the temperature control device with the central portion of the tube through the other opposing lateral side, thereby allowing the temperature control device to apply pressure to that central portion of the tube and thus allowing exposure of the central portion of the tube to the temperature control device. [Figure 5]Figure 5 shows an embodiment of a sample cartridge according to the present invention, which is brought into contact with one of two sides by a temperature control device, and on the other side of the cartridge, there is an optical detection means which is located within a counter unit that is positioned opposite the temperature control unit, forming part of the counter unit, or simply being the counter unit. On the left side (panel a), there is a cross-section, and on the right side (panel b), there is an overall view of the arrangement of the sample cartridge in the device for incubating a dispersion of particles, cells, or droplets. The particles, cells, or droplets are diffused across a flat tube, preferably as a single layer, and the tube is pressed against the opposing surface of the counter unit by the temperature control unit. [Figure 6] Figure 6 shows an embodiment of a device for incubating dispersions of particles, cells, or droplets according to the present invention. The device is shown to have a housing into which a sample cartridge as defined above is inserted, and a temperature control unit is pressed against it from one of the sides of the mounting frame. Also shown is an optical detection means located on the other side of the cartridge. [Figure 7] Figure 7 shows an exemplary cartridge embodiment according to the present invention, which includes a mounting frame, a deformable permeable tube with two opposing open ends serving as an inlet and outlet, means for reversibly closing and sealing the tube at one end in the form of a clamp, and further means for reversibly closing and sealing the orifice of the mounting frame in the form of two screw caps. In this embodiment, one or both of the screw caps are given an internal gap volume that allows them to receive and retain the liquid that has passed through means for retaining particles in the tube. [Figure 8] Figure 8 shows an exemplary cartridge embodiment with a clamp and a tube having a circular cross-section in the open position (right). Once the clamp is moved to the closed position, the tube has a circular cross-section at one end and a flat, non-circular cross-section at the other end, thus adopting a wedge shape (right). [Figure 9] Figure 9 shows an embodiment of a sample cartridge in which the upper threaded cap has been removed, while the lower threaded cap remains on the lower orifice. A pipette tip containing the liquid to be applied to the sample cartridge is shown at the top, and such liquid also contains particles. The sample is therefore applied to the cartridge and to the open upper orifice. The dispersion passes through the tube, thereby the liquid is collected in the lower cap, while particulate matter is retained in the tube by means for retaining the particles. This is an effective and convenient method for separating particles from the liquid in the dispersion. [Figure 10]Figure 10 shows an embodiment similar to Figure 8, but also demonstrates that the sample cartridge may be centrifuged after a dispersion of particles has been applied to the tube. Such centrifugation accelerates the separation of particles from the liquid. Subsequently, the lower screw cap of the tube containing the liquid that has passed through the filter is removed, and the liquid may be discarded or otherwise handled. The clamp at the end facing the means for containment (e.g., the filter) is lowered, and thus the tube is closed at such an end. Subsequently, the cartridge may be positioned upside down relative to its previous orientation and further centrifuged, so that the particles are removed from the filter and transferred back into the internal space of the tube, where they may be further resuspended in another liquid, such as a non-aqueous liquid, and accumulated at the closed clamp end of the tube. Subsequently, the sample cartridge may be introduced into a device for incubating a dispersion of particles, etc., according to the present invention, which comprises means for pressing the tube against a permeable planar substrate that forms part of the sample cartridge and acts as an opposing surface, such as a temperature control unit / surface. The tube is pressed against its surface by moving a means for pressing against the tube. Such means may be, for example, a temperature control unit having a temperature control surface adapted to be heated or cooled through the temperature control surface. To adjust the appropriate height of the internal space of the tube, predetermined spacers located on the opposing unit, on the temperature control unit, or both may be provided. Depending on the amount of pressure applied and the height of the spacers, the particles in the tube may be arranged in any desired manner, for example, a monolayer of particles may be established. A desired biochemical reaction or incubation may then occur, for example, a thermal incubation step or some of the steps thereof may be performed, such a step may lead to an optically detectable signal on or in the particles, which may then be analyzed by subsequently, for example, optically scanning the tube using optical detection means and detecting the generated signal. [Figure 11]Figure 11 shows a light transmission image of diffuse particles with a particle diameter of approximately 35 μm (panel a), and panel b shows a fluorescence image of a compressed tube with diffuse particles after a temperature incubation step, e.g., thermal cycling. The image size is 2 × 6 cm², and the clamp area can be seen on the left side of the image. [Figure 12a] Figure 12 shows an embodiment of a sample cartridge according to the present invention, where panel a shows a side view of the sample cartridge, panel b shows a top view and a cross-sectional side view thereof, and panel c shows a side view thereof, and panel c also shows part of an embodiment of a device for incubating dispersions of particles, cells, or droplets, including a temperature control unit and optical detection means. Reference numerals are included. [Figure 12b] Figure 12 shows an embodiment of a sample cartridge according to the present invention, where panel a shows a side view of the sample cartridge, panel b shows a top view and a cross-sectional side view thereof, and panel c shows a side view thereof, and panel c also shows part of an embodiment of a device for incubating dispersions of particles, cells, or droplets, including a temperature control unit and optical detection means. Reference numerals are included. [Figure 12c] Figure 12 shows an embodiment of a sample cartridge according to the present invention, where panel a shows a side view of the sample cartridge, panel b shows a top view and a cross-sectional side view thereof, and panel c shows a side view thereof, and panel c also shows part of an embodiment of a device for incubating dispersions of particles, cells, or droplets, including a temperature control unit and optical detection means. Reference numerals are included. [Modes for carrying out the invention]

[0008] In a first aspect, the present invention relates to a sample cartridge (100) for incubating and / or analyzing a dispersion (200) of particles, cells, or droplets, in particular a suspension (210) of particles or cells, or an emulsion (220) of droplets, and / or for carrying out a biochemical reaction with or in such a dispersion, the cartridge is A deformable permeable tube (110), Two opposing open ends (111, 112), each serving as an inlet and an outlet. A deformable permeable tube (110) having Equipped with, The tube is adapted to receive a dispersion of particles, cells, or droplets, particularly a suspension of particles or cells, or an emulsion of droplets, and within the internal space (113) of the tube, the internal space is lined by one wall (114'') or several walls (114, 114', 114'') of the tube, and the tube is configured such that when the internal space of the tube receives the suspension, emulsion, or dispersion, it has a circular or oval cross-section, and when the internal space is pressed against a surface, it has a flat, non-circular cross-section. The cartridge further comprises means (120) for retaining the particles, cells, or droplets within the deformable permeable tube, the means for retaining the particles being located at one or both ends of the tube, and the means for retaining the particles is preferably a filter, membrane, grid, mesh, sieve, or other structure that retains the particles while allowing the passage of a liquid through it.

[0009] In one embodiment, the deformable permeable tube has a single wall (114'''), and the internal space (113) is lined by the single wall.

[0010] In one embodiment, the sample cartridge further comprises means (130) for reversibly closing and sealing the deformable permeable tube at one or both of the opposing ends (111, 112), preferably the means for reversibly sealing being a clamp, or, if both ends are sealed, a pair of clamps located at the opposing ends.

[0011] In one embodiment, the sample cartridge further comprises a mounting frame (140), the mounting frame (140) connected to and holding the tube at the opposing ends of the tube, and configured to allow the addition of a substance, such as a liquid or solid or a mixture thereof, to the internal space of the tube through one of the ends which serves as an inlet, and / or the removal of a substance, such as a liquid or solid or a mixture thereof, from the internal space of the tube through the other end which serves as an outlet.

[0012] In one embodiment, the mounting frame (140) has first and second lateral sides (141, 142) located opposite each other, one of which is formed of a permeable planar substrate configured to act as an opposing surface (321) against which the tube can be pressed, and the mounting frame is configured such that the other lateral side of the mounting frame allows physical contact of the temperature control device to the central portion of the tube through the other opposing lateral side, and allows pressure to be applied by the temperature control device to the central portion of the tube through the other opposing lateral side, preferably against the opposing surface of the permeable planar substrate, thereby allowing exposure of the central portion (115) of the tube to the temperature control device.

[0013] In one embodiment, the mounting frame (140) is further preferably configured to allow analysis of the central portion (115) of the tube by optical detection means through one of the lateral sides of the mounting frame, more preferably through the lateral side formed by the transparent planar substrate as defined above.

[0014] In one embodiment, the central portion (115) of the tube is the portion that is closed and sealed by the means (130) for reversibly closing and sealing the deformable permeable tube.

[0015] In one embodiment, the mounting frame has a longitudinal axis (143) aligned with the longitudinal axis (116) of the pipe, and the mounting frame comprises two opposing longitudinal ends (144, 145), each of which has an orifice (146, 147) that fluidly connects to the opposing ends (111, 112) of the pipe, each of which serves as the inlet and outlet of the pipe, and each of the orifices is sealable and preferably comprises means (148, 148') for reversibly closing and sealing such orifice, such as a cap, tap, plug, stopper, or screw cap.

[0016] In one embodiment, the tube (110) is mounted within a mounting frame (140) such that the opposing open end of the tube is attached to or in contact with the opposing longitudinal end of the mounting frame, the mounting frame encompassing a space (149) through which the tube extends, such space being configured to allow exposure to or contact of the central portion (115) of the tube with a temperature control device and / or to allow analysis of the central portion of the tube by optical detection means.

[0017] In one embodiment, the walls (114, 114', 114'') of the tube, preferably the single wall (114'''), have a thickness in the range of 1 μm to 1,000 μm, preferably 20 μm to 200 μm, more preferably 50 μm to 150 μm, and / or the diameter of the internal space is in the range of 0.1 cm to 5 cm when it has a circular or oval cross-section, and / or the height of the internal space of the tube is in the range of 5 μm to 500 μm, preferably 5 μm to 200 μm, more preferably 10 μm to 150 μm when it has a flat, non-circular cross-section.

[0018] In one embodiment, the tube is a seamless tube and preferably has no edges, in particular any outer edges, such as those that would result from a sealing process. This has the advantage that gas will not be trapped by any such edges. In one embodiment, the tube does not have nucleic acids adhering to the inside of the one or more walls lining the internal space of the tube.

[0019] In one embodiment, the pipe is not a branched pipe or a pipe having one or more bends or constrictions therein. In one embodiment, the pipe does not have multiple chambers or reservoirs or compartments or internal spaces.

[0020] In one embodiment, the pipe is a pipe having a single internal space and a longitudinal axis, and the opposing open ends of the pipe are arranged such that they are aligned with each other along the longitudinal axis.

[0021] In one embodiment, the opposing open ends are of the same size, and preferably they have openings of substantially the same diameter or differing in diameter by only 10% or less.

[0022] In one embodiment, the deformable permeable tube is permeable in the range of 250 nm to 950 nm, preferably in the range of 400 nm to 600 nm, and / or in the range of 450 nm to 650 nm, and / or in the range of 500 nm to 700 nm, and / or in the range of 550 nm to 750 nm, and / or in the range of 600 nm to 800 nm, and is made from a material that, if present, allows analysis of any contents in the internal space of the tube using optical spectroscopy and / or imaging, preferably the material is styrene-butadiene rubber, silicone rubber, polyvinyl butyral, polyurethane, polyisobutylene, polyhydroxybutyrate, polyhydroxyalkanoate, polyether block amide, rubber, gum arabic, isoprene rubber, fluororubber, ethylene vinyl acetate copolymer, ethylene-propylene diene-rubber copolymer, ethylene-ethyl acrylate copolymer, chloro Selected from polypropylene rubber, ethyl rubber, butadiene rubber, acrylonitrile-methyl methacrylate copolymer, acrylonitrile-chlorinate-polyethylene-styrene copolymer, acrylonitrile-butadiene-acrylate copolymer, polyester (PES), polyethylene terephthalate (PET), polyethylene (PE), high-density polyethylene (HDPE), polyvinyl chloride (PVC), polyvinylidene chloride (PVDC), low-density polyethylene (LDPE), polypropylene (PP), polystyrene (PS), high-impact polystyrene (HIPS), polyamide (PA), acrylonitrile butadiene styrene (ABS), polyethylene / acrylonitrile butadiene styrene (PE / ABS), polycarbonate (PC), polycarbonate / acrylonitrile butadiene styrene (PC / ABS), polyurethane (PU), and any combination or copolymer of the foregoing.

[0023] In a further aspect, the present invention also relates to the use of sample cartridges according to the present invention for generating and / or processing dispersions of particles, cells, or droplets, particularly suspensions of particles or cells, or emulsions of droplets.

[0024] In one embodiment, the process is one or more of the following activities: incubating the dispersion of particles, cells, or droplets; carrying out a biochemical reaction with the dispersion of particles, cells, or droplets; binding one or more analytes to the particles, cells, or droplets, and then removing any unbound analytes and other unbound substances from the particles, cells, or droplets; exchanging the liquid phase of the dispersion; and analyzing the dispersion of particles, cells, or droplets.

[0025] In one embodiment, the use is The steps include filling the tube with a dispersion of particles, cells, or droplets, particularly a suspension of particles or cells, and optionally one or more additional reagents. The steps include optionally arranging the particles, cells, or droplets in a single layer, The steps include carrying out a biochemical reaction in the tube and / or incubating the tube under one or more defined reaction conditions, in particular under one or more temperature conditions, The steps include analyzing the results of such biochemical reactions and / or such incubations, Includes.

[0026] In one embodiment, the biochemical reaction is not a sequencing process and / or does not involve the use of any nucleic acids adhering to the inside of the one or more walls lining the internal space of the tube.

[0027] In one embodiment, the use is A step of filling a tube with a dispersion of particles, cells, or droplets, wherein the dispersion comprises a first liquid, preferably a first aqueous liquid. The steps include removing the first liquid from the particles, cells, or droplets in the tube by, for example, centrifugal force, gravity, or suction, by allowing the first liquid to pass through the means for containing the particles, cells, or droplets while the particles, cells, or droplets are contained by the means for containing the particles, cells, or droplets; The steps include adding a second liquid containing the analyte, preferably a second aqueous liquid, to the tube, A step of incubating the particles, cells, or droplets in the second liquid to enable or facilitate the binding of the analyte to the particles, cells, or droplets, The steps include: washing the particles, cells, or droplets to remove the second liquid from the particles, cells, or droplets in the tube, for example by centrifugation, gravity, or suction, by allowing the second liquid to pass through the means for retaining while the particles, cells, or droplets are retained by the means for retaining, and optionally removing any unbound analytes and unbound substances from the particles, cells, or droplets; The steps include: resuspending the particles in a third liquid, preferably a non-aqueous liquid, by adding such a third liquid to the tube; The steps include optionally arranging the particles, cells, or droplets in a single layer, The steps include carrying out a biochemical reaction in the tube and / or incubating the tube under one or more defined reaction conditions, in particular under one or more temperature conditions, The steps include analyzing the results of such biochemical reactions and / or such incubations, Includes.

[0028] In one embodiment, the biochemical reaction is not a sequencing process and / or does not involve the use of any nucleic acids adhering to the inside of the one or more walls lining the internal space of the tube.

[0029] In a further aspect, the present invention also relates to a method for producing droplet dispersions, in particular droplet emulsions, the method comprising the steps of providing a cartridge according to the present invention as defined above, and mixing an aqueous phase and an oily liquid phase within the tube of the cartridge, thereby producing a dispersion, in particular droplet emulsion.

[0030] In a further aspect, the present invention also relates to a method for producing a solid or semi-solid, for example, a dispersion of gel particles, the method comprising the steps of providing a cartridge according to the present invention as defined above, and mixing an aqueous phase and an oily liquid phase in the tube of the cartridge, thereby producing a dispersion, in particular an emulsion of droplets, one of the phases in addition containing any solid particles or one or more components capable of forming a gel or solid in response to changing at least one environmental condition around the components, and if one of the phases in addition contains one or more components capable of forming a gel or solid, the method in addition includes the step of inducing the formation of a gel or solid by changing the at least one environmental condition around the components, thereby converting the droplets into particles and producing a dispersion of particles in the tube.

[0031] In one embodiment, the at least one environmental condition is selected from temperature, pH, pressure, light within a defined wavelength range, ultrasound, and the presence of polymerization-inducing chemicals.

[0032] In a further aspect, the present invention also relates to a device (300) for incubating and / or carrying out biochemical reactions with dispersions of particles, cells, or droplets, particularly suspensions of particles or cells, or emulsions of droplets, the device A sample cartridge (100) according to the present invention as defined above, A temperature control unit (310) having a temperature control surface (311) and adapted to heat and / or cool through the temperature control surface, Equipped with, The device is configured such that the tube of the sample cartridge, particularly the central portion (115) of the tube, can be brought into contact with the temperature control surface (311) using one of its lateral sides (141, 142), or is in contact with and can be pressed by or against the temperature control surface (311), thereby the tube is deformed when pressed by or against the temperature control surface, and the internal space (113) of the pressed tube has a flat, non-circular cross-section.

[0033] In one embodiment, the device further comprises a counter unit (320) positioned opposite the temperature control unit (310) at a certain distance therefrom, and having a counter surface (321) facing the temperature control surface (311), wherein the counter unit is either a permeable planar substrate that, if present in the cartridge, forms one of the lateral sides (141, 142) of the cartridge and is configured to act as a counter surface (321) against which the tube can be pressed, or the counter unit (320) is a separate component that does not form part of the cartridge and is provided separately in the device (300) apart from the cartridge, wherein the separate component is preferably configured to be operable to apply pressure through the counter surface (321) on the tube in contact with the temperature control surface (311), or operable to be positioned at a defined distance from the sample cartridge (109).

[0034] In one embodiment, one or both of the temperature control surface or the opposing surface are preferably transparent in the range of 250 nm to 950 nm, and preferably transparent in the range of 400 nm to 600 nm and / or in the range of 450 nm to 650 nm and / or in the range of 500 nm to 700 nm and / or in the range of 550 nm to 750 nm and / or in the range of 600 nm to 800 nm.

[0035] In one embodiment, the temperature control unit has a receiving portion (312) for receiving the tube, the receiving portion enabling the tube to be fixed on the temperature control surface (311).

[0036] In one embodiment, the device further, The opposing unit (320) comprises one or more spacers (330, 330') located on the opposing surface (321) or on the temperature control unit (310), preferably on the temperature control surface (311), wherein the spacers (330, 330') have a height represented by the following formula: H S = 2 × T + H CS In the formula, H S = height of the spacer, T = wall thickness of the deformable permeable tube, H CS = The height of the internal space of the tube when it has a flat, non-circular cross-section, and if there are several spacers (330, 330'), the height of each spacer is the same H S And preferably, H S The range is from 7 μm to 2,500 μm.

[0037] In one embodiment, the device further comprises an optical detection means (340) configured to detect and / or analyze the contents of the internal space (113) of the tube using optical spectroscopy and / or imaging, preferably such detection and / or analysis is performed using a beam path that travels through the central portion (115) of the tube and one or both of the temperature control surface (311) or the opposing surface (321).

[0038] In one embodiment, the device comprises a plurality of sample cartridges (100, 100', 100'') as defined above.

[0039] In a further aspect, the present invention also relates to the use of the device according to the present invention for generating and / or processing dispersions of particles, cells, or droplets, in particular suspensions of particles or cells, or emulsions of droplets, the use of which is carried out with a sample cartridge according to the present invention as defined above and as defined above.

[0040] The inventors have surprisingly found that the use of an open-end deformable permeable tube configured within a cartridge makes it possible to achieve the desired results. The tube is adapted to receive a dispersion of particles, cells, or droplets in the internal space of the tube formed by one, two, or several walls of the tube, and the tube is configured such that the internal space of the tube has a circular or oval cross-section when it is receiving a suspension or emulsion or dispersion, and the internal space of the tube has a flat non-circular cross-section when the tube is pressed against a surface. The sample cartridge according to the present invention further comprises means for retaining particles, cells, or droplets within the deformable permeable tube while allowing free passage or transfer of the liquid through which such means are located. The means for retaining particles are typically located at one or both ends of the tube. Preferably, such means are a filter, membrane, grid, mesh, sieve, or other structure that allows the passage of the liquid through which it is located while retaining the particles. The tube is deformable within the range of having a circular or oval cross-section when it is filled and after it has been filled with a liquid such as a dispersion of particles, cells, or droplets, and within the range of having a flat, non-circular cross-section when the tube receives the liquid and is pressed against a surface in such a filled state. This allows for a reaction space that is available to have one dimension small enough to allow for rapid heat transfer from the temperature-controlled surface when the tube is pressed against such a surface. By appropriately selecting and defining the way in which the tube is pressed against a surface, an optimized reaction space can be generated. For example, if the tube is pressed against a surface in only one portion, e.g., at one end, the internal space of the tube may adopt a wedge shape that allows for the directing and removal of any gas / air that may be trapped during the tube filling process. This aeration process can be assisted by gravity. When the tube is subsequently pressed against a surface over a substantial portion of the tube, e.g., over its substantial central portion, the tube will adopt the aforementioned flat, non-circular cross-section along the entire length of such central portion.The internal space of the compressed tube may then have a height just large enough to accommodate a single layer of particles, cells, or droplets. Thus, in such embodiments of the sample cartridge and method according to the present invention, the particles, cells, or droplets are arranged in a single layer. This is advantageous in particular, given that such an arrangement then allows for both fast and efficient heat transfer and analysis of individual particles, cells, or droplets without any overlap with other particles. In one embodiment, providing a suitable height for the internal space depends on various factors, including the pressure applied to the tube, the size of the particles, the elasticity of the tube if present, the height of the spacer provided if present, and others. Thus, in one embodiment, the cartridge according to the present invention may be used to arrange a sample containing a dispersion of particles such that the particles form a single layer. In a preferred embodiment, the deformable permeable tube has a single wall, and the internal space is lined by a single wall. This has the advantage that gas formation at the edges, for example between different parts of the tube, is reduced because there are no edges. In one embodiment, the tube is a seamless tube and preferably has no edges, in particular any outer edges, such as those that would result from a sealing process. This has the advantage that gas will not be trapped in any such edges. In one embodiment, the tube does not have nucleic acids adhering to the inside of the one or more walls lining the internal space of the tube. In one embodiment, the tube is an elastic tube and is made from an elastic material, preferably an elastomer material. In one embodiment, the tube is made from a material whose glass transition temperature is below the temperature at which the material is used. In another embodiment, the tube is a plastic tube and is made from a plastic material, preferably a thermoplastic material.In one embodiment, the tube is made of a polymer material, preferably styrene-butadiene rubber, silicone rubber, polyvinyl butyral, polyurethane, polyisobutylene, polyhydroxybutyrate, polyhydroxyalkanoate, polyether block amide, rubber, gum arabic, isoprene rubber, fluororubber, ethylene vinyl acetate copolymer, ethylene-propylenediene-rubber copolymer, ethylene-ethyl acrylate copolymer, chloroprene rubber, ethyl rubber, butadiene rubber, acrylonitrile-methyl methacrylate copolymer, acrylonitrile-chlorinate-polyethylene-styrene copolymer, acrylonitrile-butadiene-acrylate copolymer, polyester It is made from polymer materials selected from terephthalate (PES), polyethylene terephthalate (PET), polyethylene (PE), high-density polyethylene (HDPE), polyvinyl chloride (PVC), polyvinylidene chloride (PVDC), low-density polyethylene (LDPE), polypropylene (PP), polystyrene (PS), high-impact polystyrene (HIPS), polyamide (PA), acrylonitrile butadiene styrene (ABS), polyethylene / acrylonitrile butadiene styrene (PE / ABS), polycarbonate (PC), polycarbonate / acrylonitrile butadiene styrene (PC / ABS), polyurethane (PU), and any combination or copolymer of the aforementioned.

[0041] In one embodiment, the sample cartridge according to the present invention further comprises means for reversibly closing and sealing a deformable permeable tube at one or both of the opposing ends. Such means for reversibly closing and sealing the permeable tube at one or both ends may be arranged such that one end is closed and sealed, while the other end remains temporarily open, and thus the volume of the internal space is adapted. Excess liquid sample or unwanted gas / air may be removed through the open end. In a preferred embodiment, means for reversibly closing and sealing the deformable permeable tube at both opposing ends are present. In such an embodiment, the second means may also provide a defined internal space which is an optimized reaction space, allowing the deformable elastic permeable tube to be subsequently closed and sealed at both ends. Preferably, the means for reversibly closing and sealing the deformable permeable tube are configured to withstand pressure and / or heat when they are closed / sealed. This has the advantage that the sample in the internal space can be exposed to temperatures >80°C, e.g., 81°C, 82°C, 83°C, 84°C, 85°C, 86°C, 87°C, 88°C, 89°C, 90°C, 91°C, 92°C, 93°C, 94°C, 95°C, 96°C, 97°C, 98°C, 99°C, 100°C, etc., without the risk of the tube losing the sample from the internal space, and the sample cartridge can be exposed to external forces, such as during centrifugation. In some special cases, the internal space may be heated to temperatures up to >120°C, which may be beneficial for ultrafast thermal cycling of the sample.

[0042] In one embodiment, the means for reversibly closing and sealing is a clamp or a pair of clamps located at one or both of the opposing ends of the pipe.

[0043] In one embodiment, the sample cartridge according to the present invention further comprises a mounting frame connected to and holding the tube at the opposing ends of the tube. Such a mounting frame is configured to allow the addition of a substance, such as a liquid or solid or a mixture thereof, to the internal space of the tube through one of the ends which acts as an inlet, and / or the removal of a substance, such as a liquid or solid or a mixture thereof, from the internal space of the tube through the other end which acts as an outlet.

[0044] Means for containing particles located at one or both ends of the tube are either located inside the tube or outside the tube but at the end of the tube within the sample cartridge.

[0045] The mounting frame for the sample cartridge provides stability to the sample cartridge and allows for protection of the tube while enabling controlled and directed access to such tube. In one embodiment, such a mounting frame is made from a material that provides such mechanical stability. A number of suitable materials such as plastic, metal, wood, glass, or ceramic may be envisioned. In one embodiment, the mounting frame has first and second lateral sides positioned opposite each other, one of which is formed by a permeable planar substrate, preferably configured to act as an opposing surface against which the tube can be pressed. The mounting frame is configured such that the other lateral side of the mounting frame, i.e., the side not formed by the permeable planar substrate, remains open and allows exposure of the central portion of the tube to a temperature control device that is not part of the sample cartridge. This lateral side allows for physical contact of the temperature control device with the central portion of the tube and the application of pressure to the central portion of the tube by such temperature control device. For example, the temperature control device may press the tube against the opposing surface of the permeable planar substrate. The mounting frame is further configured to allow analysis of the central portion of the tube by optical detection means (such optical detection means again not form part of the sample cartridge). Such optical detection / analysis may occur through one of the two lateral sides of the mounting frame, preferably through the side formed by a transparent planar substrate, as further defined above.

[0046] In one embodiment, the central portion of the tube to be analyzed is the portion that is closed and sealed by means for reversibly closing and sealing a deformable permeable tube. As used herein, “analysis of the central portion of the tube” means analysis of the internal space of the tube within such a central portion. The analysis may be performed by any preferred means, e.g., optical detection means or imaging means enabling optical analysis or imaging. In one embodiment, the mounting frame is configured to allow centrifugation of the sample cartridge. Preferably, this is enabled by fitting and shaping the sample cartridge to allow insertion and fitting into a centrifugation tube and / or centrifugation rotor. In one embodiment, the sample cartridge is capable of withstanding centrifugal accelerations up to 10,000 g. In one embodiment, the sample cartridge is “centrifugable.” Such “centrifugable” means the ability of such a sample cartridge to be centrifuged without becoming damaged, permanently deformed, or otherwise undesirably affected in an undesirable manner. In one embodiment, the sample cartridge is suitable for and intended to be centrifuged.

[0047] In a preferred embodiment, the sample cartridge mounting frame has a longitudinal axis aligned with the longitudinal axis of the deformable permeable tube. In such an embodiment, the sample cartridge mounting frame comprises two opposing longitudinal ends, each having an orifice, such orifice fluidly connected to the adjacent individual ends of the tube, namely, the end that serves as the inlet and the end that serves as the outlet of the tube. In such an embodiment, each of these orifices is sealable and preferably comprises means for reversibly closing and sealing such orifices. Such means for reversibly closing and sealing such orifices may be any preferred means, e.g., a cap, tap, plug, stopper, or screw cap. The advantage of such sealable orifices is again that this increases the stability and rigidity of the cartridge and facilitates centrifugal separation of such cartridge. Furthermore, the means for reversibly closing and sealing the orifices may be configured such that they themselves are capable of receiving a certain volume of liquid, for example, when the sample cartridge is centrifuged to separate particles of a dispersion added to the internal space of the tube from the liquid. For example, such means, e.g., a cap, screw cap, tab, plug, or stopper, may provide an internal gap volume capable of receiving such liquid. In response to the application of a force such as gravity or centrifugal force to the cartridge, particles, cells, or droplets are retained by means for retaining them within the sample cartridge, while liquids pass through such means and are received by means for reversibly closing and sealing the orifice, e.g., a cap. Once received within such means for reversibly closing and sealing, the liquid can be removed, discarded, or otherwise handled.

[0048] In one embodiment, a deformable permeable tube is mounted within the cartridge mounting frame such that opposing open ends of the tube are attached to or in contact with opposing longitudinal ends of the mounting frame. In such embodiments, means for containing particles, cells, or droplets are located just inside the tube, at one or both ends of the tube, or they are located outside the tube at opposing longitudinal ends of the mounting frame. In one embodiment, the mounting frame encompasses a space through which the tube extends longitudinally, and such space is configured to allow exposure to or contact of the central portion of the tube with a temperature control device (which does not form part of the sample cartridge) and / or analysis of the central portion of the tube by optical detection means (again, such optical detection means does not form part of the sample cartridge).

[0049] In one embodiment, the wall of the tube, preferably a single wall of the tube, has a thickness in the range of 1 μm to 1,000 μm, preferably 20 μm to 200 μm, more preferably 50 μm to 150 μm. In one embodiment, the diameter of the internal space is in the range of 0.1 cm to 5 cm when the tube has a circular or oval cross-section. In one embodiment, when the tube has a flat non-circular cross-section, the height of the internal space of the tube is in the range of 5 μm to 500 μm, preferably 5 μm to 200 μm, more preferably 10 μm to 150 μm. In one embodiment, such height of the internal space of the tube is selected to match the dimensions of a particle, cell, or droplet that is part of the sample being analyzed / processed. In one embodiment, when the height of the internal space of the tube has a flat non-circular cross-section match, it is the same as or approximately the same as the height of a single particle, cell, or droplet that forms part of the sample, but it may also be 1 to 10 μm greater than the height of the single particle, cell, or droplet. The correspondence between the height of a single particle and the size of the internal space allows for efficient processing and / or analysis of a single layer of particles, cells, or droplets within the internal space of the tube. Analysis and / or processing can be performed at the bulk level (e.g., obtaining an analytical image from the tube) or at the single-particle level (e.g., manipulating a single particle with a laser beam). In one embodiment, the permeable tube is fabricated from a material that is transparent to light in the range of 250 nm to 950 nm. Thus, it can be transparent over the entire range, or it can be transparent within its partial range. Depending on the sample and the method of analyzing such a sample, different ranges of transparency may be preferable. In one embodiment, the material is transparent in the range of 400 nm to 600 nm, and / or 450 nm to 650 nm, and / or 500 nm to 700 nm, and / or 550 nm to 750 nm, and / or 600 nm to 800 nm. At least the combination of materials needs to be transparent so that the selected optical detection principle can be implemented. This can lead to situations, for example, in fluorescence detection, where the material is either transparent or semi-transparent to only two wavelengths.Furthermore, the side of the tube not facing the optical detection means or optical detection unit may be treated to alter its optical properties in order to improve the optical detection, for example, by coloring it black. In one embodiment, the material of the deformable permeable tube allows for analysis of any contents within the internal space of the tube using optical spectroscopy and / or imaging, if present. In one embodiment, the tube is made from a material that is ductile and / or plastic and / or elastic and / or thermoformable and / or thermoplastic elastomer, or it may have any other flexible properties to have the ability to form a “detection chamber”. This applies to almost all plastic materials with a temperature at least above the glass transition temperature. In one embodiment, the tube is made of styrene-butadiene rubber, silicone rubber, polyvinyl butyral, polyurethane, polyisobutylene, polyhydroxybutyrate, polyhydroxyalkanoate, polyether block amide, rubber, gum arabic, isoprene rubber, fluororubber, ethylene vinyl acetate copolymer, ethylene-propylenediene-rubber copolymer, ethylene-ethyl acrylate copolymer, chloroprene rubber, ethyl rubber, butadiene rubber, acrylonitrile-methyl methacrylate copolymer, acrylonitrile-chlorinate-polyethylene-styrene copolymer, acrylonitrile-butadiene-acrylate copolymer, polyester (PE) S) is made from materials selected from polyethylene terephthalate (PET), polyethylene (PE), high-density polyethylene (HDPE), polyvinyl chloride (PVC), polyvinylidene chloride (PVDC), low-density polyethylene (LDPE), polypropylene (PP), polystyrene (PS), high-impact polystyrene (HIPS), polyamide (PA), acrylonitrile butadiene styrene (ABS), polyethylene / acrylonitrile butadiene styrene (PE / ABS), polycarbonate (PC), polycarbonate / acrylonitrile butadiene styrene (PC / ABS), polyurethane (PU), and any combination or copolymer of the aforementioned.

[0050] The present invention also relates to the use of a sample cartridge as defined above for generating and / or processing dispersions of particles, cells, or droplets, particularly suspensions of particles or cells, or emulsions of droplets. Similarly, the present invention also relates to a method for generating and / or processing dispersions of particles, cells, or droplets, particularly suspensions of particles or cells, or emulsions of droplets, in which a sample cartridge as defined above is used.

[0051] In one embodiment, the process is one or more of the following activities: incubating the dispersion of particles, cells, or droplets; carrying out a biochemical reaction with the dispersion of particles, cells, or droplets; binding one or more analytes to the particles, cells, or droplets, and then removing any unbound analytes and other unbound substances from the particles, cells, or droplets; exchanging the liquid phase of the dispersion; and / or analyzing the dispersion of particles, cells, or droplets.

[0052] In one embodiment, the use is The steps include filling the tube with a dispersion of particles, cells, or droplets, particularly a suspension of particles or cells, and optionally one or more additional reagents. The steps include carrying out a biochemical reaction in the tube and / or incubating the tube under one or more defined reaction conditions, in particular under one or more temperature conditions, The steps include analyzing the results of such biochemical reactions and / or such incubations, Includes.

[0053] In one embodiment, use is preferably, A step of filling a tube with a dispersion of particles, cells, or droplets, wherein the dispersion comprises a first liquid, preferably a first aqueous liquid. The steps include removing the first liquid from the particles, cells, or droplets in the tube by, for example, centrifugal force, gravity, or suction, by allowing the first liquid to pass through the means for containing the particles, cells, or droplets while the particles, cells, or droplets are contained by the means for containing the particles, cells, or droplets; The steps include adding a second liquid containing the analyte, preferably a second aqueous liquid, to the tube, A step of incubating the particles, cells, or droplets in the second liquid to enable or facilitate the binding of the analyte to the particles, cells, or droplets, The steps include: washing the particles, cells, or droplets to remove the second liquid from the particles, cells, or droplets in the tube, for example by centrifugation, gravity, or suction, by allowing the second liquid to pass through the means for retaining while the particles, cells, or droplets are retained by the means for retaining, and optionally removing any unbound analytes and unbound substances from the particles, cells, or droplets; The steps include: resuspending the particles in a third liquid, preferably a non-aqueous liquid, by adding such a third liquid to the tube; The steps include carrying out a biochemical reaction in the tube and / or incubating the tube under one or more defined reaction conditions, in particular under one or more temperature conditions, The steps include analyzing the results of such biochemical reactions and / or such incubations, Includes.

[0054] With respect to dispersions of particles, cells, or droplets, and any suitable particles, cells, or droplets may be used in conjunction with the sample cartridge according to the present invention, insofar as they enable the desired processing / reaction to be carried out. Such particles may be microbeads capable of attaching capture molecules. Such particles are, in principle, publicly known. Preferred examples are disclosed, for example, in Microfluidic Methods for Molecular Biology, Lu & Verbridge Editors, Springer International Publishing Switzerland 2016. Further preferred examples are disclosed, for example, in the pending European Patent Application No. 16207455.3 filed on 30 December 2016. These particles are examples of pre-processed microparticles for carrying out the digital detection of any analyte in a sample, having a surface, accepting aqueous solutions, and containing gap volumes that are dispersible in a non-aqueous medium.

[0055] In a further aspect, the sample cartridge according to the present invention can also be used to generate droplet emulsions or particle dispersions. Thus, in one embodiment, the present invention also relates to the use of a sample cartridge as defined above for generating droplet emulsions or particle dispersions. This also relates to a method for generating droplet emulsions or particle dispersions in which a sample cartridge as defined above is used. Such generation may be carried out as an independent process or prior to any incubation and other processes such as processes relating to the detection or analysis of reaction products. Thus, when it is further described above that particles, cells, or droplet dispersions are filled into the tube (of the cartridge), this also means that it includes the possibility of generating (from scratch) particles, cells, or droplet dispersions such as droplet emulsions or particle dispersions in the tube. In one embodiment, the sample cartridge according to the present invention can be used to generate aqueous droplets, i.e., water-in-oil emulsions. In another embodiment, the sample cartridge according to the present invention can be used to generate oily droplets, i.e., oil-in-water emulsions. To generate a stable emulsion, emulsifiers may be used. Such emulsifiers belong to the category of surfactant materials with a suitable hydrophilic-lipophilic balance (HLB) value for producing either water-in-oil (W / O) or oil-in-water (O / W) emulsions. Emulsifiers can generally be classified according to the properties of their hydrophilic heads and are grouped into anionic, cationic, zwitterionic, and nonionic surfactants. Typically, a phase with better solubility for the emulsifier is used as the mobile phase.

[0056] In one embodiment, with respect to aqueous droplet generation (water-in-oil emulsion), the tube of the sample cartridge is clamped at an open end and filled through the opposing open end with a defined volume of oil phase liquid that serves as the mobile phase, such as commercially available mineral oil, paraffin oil, or industrial fluid (e.g., fluorocarbon or hydrofluoroether), or a suitable organic solvent containing a suitable emulsifier as optional. Subsequently, in this embodiment, a defined volume of aqueous solution representing a dispersed phase containing the substance / solute intended to be encapsulated in the droplet is added to the tube. In one embodiment, the ratio of the defined volume of oil phase liquid to the defined volume of aqueous solution is in the range of 1.2:1 to 100:1, preferably 2:1 to 10:1. The ratio of the mobile relative emulsion phase can vary and depends on the substance employed for the mobile phase, the dispersed phase, and the surfactant used to stabilize the emulsion. Multiple protocols can be found in textbooks such as Tadros, Tharwat F., Emulsions, Formation, Stability, Industrial Applications, ISBN 978-3-11-045224-2. Substances that may be contained in the aqueous solution may be amplification reagents such as PCR reagents, and may further include amplification targets, detection reagents, emulsifiers, etc. Typical protocols for emulsion PCR are publicly known, and examples are described in Williams et al., Nature Methods 3(7):545-550, 2006. After closing the cartridge inlet, droplets are generated by agitating the tubing, which can be done directly by any preferred means such as repeatedly compressing the tubing, or by other means such as stirring, shaking, or applying ultrasound. After the completion of the droplet generation procedure, the cartridge may be treated as described above. For example, the droplet may be washed, incubated, or exposed to defined reaction conditions such as one or more temperature conditions, or a biochemical reaction may be carried out within the tube.

[0057] In addition, in one embodiment, the droplet-forming solution, in this case an aqueous solution, may contain reagents that enable the conversion of droplets into particles or capsules. In such embodiments, the generated droplets can be further converted into particles (solid or semi-solid, e.g., gel) by, for example, a gelation process and by a polymerization process. Thus, in this embodiment, the emulsion of generated droplets is subsequently further converted into a suspension of particles. For example, the droplet-forming solution, i.e., the aqueous solution, may contain a gelling substance such as agarose or gelatin that can be induced to gel, or it may contain a suitable monomer or prepolymer that can be induced to polymerize, such as bisacrylamide, and a suitable diamine together with a catalyst such as ammonium persulfate to initiate a redox reaction. Capsules and particles can be formed by well-established means such as ionotropic gelation, coacervation, interfacial polycondensation, interfacial crosslinking, in-situ polymerization, and matrix polymerization. Furthermore, multilayer techniques may be used to construct customized capsule arrays with tuned properties (layer-by-layer assembly of microcapsules and their biomedical Applications are outlined in Tong W, Song X, Gao C.; Chem Soc Rev. 2012 Sep 21;41(18):6103-24.

[0058] In such embodiments, after droplet formation, the tube and its contents, i.e., the cartridge which also includes the generated droplets, are exposed to gel-inducing or polymerization-inducing conditions. In a simple form, such gel-inducing or polymerization-inducing conditions may be a change in temperature or exposure to electromagnetic radiation in a defined wavelength range, such as UV light. The particles thus generated may be further processed as described above. For example, the particles may be washed, incubated, or exposed to defined reaction conditions such as one or more temperature conditions, or a biochemical reaction may be carried out within the tube.

[0059] By applying reagents in different proportions, an oil-in-water emulsion may be formed, comprising an aqueous phase acting as a continuous phase and an oil phase as a dispersed phase. A suitable protocol can be found, among other things, in Tadros, Tharwat F., Emulsions, Formation, Stability, Industrial Applications, ISBN 978-3-11-045224-2.

[0060] In a further aspect, the present invention also relates to a device for incubating and / or carrying out biochemical reactions with dispersions of particles, cells, or droplets, particularly suspensions of particles or cells, or emulsions of droplets, the device comprising: A sample cartridge as defined above, A temperature control unit having a temperature control surface and adapted to heat or cool through the temperature control surface, Equipped with, The device is configured such that the tube of the sample cartridge, particularly the central portion of the tube, can be brought into contact with the temperature control surface of the device and can be pressed by or against the temperature control surface, thereby the tube is deformed when pressed by or against the temperature control surface, and the internal space of the pressed tube has a flat, non-circular cross-section.

[0061] In a preferred embodiment of the device, such a device further comprises a counter unit positioned opposite the temperature control unit and having a counter surface facing the temperature control surface, wherein the counter unit, when located within the cartridge, is a permeable planar substrate configured to form one of the lateral sides of the cartridge and act as a counter surface against which the tube can be pressed, or the counter unit is a separate component that does not form part of the cartridge and is provided within the device separately from the cartridge. In one embodiment, the separate component is preferably configured to be operable to apply pressure via the counter surface on the tube in contact with the temperature control surface, or the separate component is operable to be positioned at a defined distance from the sample cartridge. Such a defined distance may be in the range of 0 μm to 10 mm. If the counter unit is positioned at a distance of 0 μm as a separate component that does not form part of the cartridge, it can be in virtual contact with the cartridge and apply pressure directly or indirectly to the tube of the cartridge. For example, if the cartridge does not have a permeable planar substrate on one of the sides of the mounting frame, the opposing unit may be in direct contact with the tube, and the temperature control surface (pressing from the other side) may apply pressure to the tube. Alternatively, if a permeable planar substrate is present within the sample cartridge, the opposing unit, as a separate component, may be in contact with such a permeable planar substrate against which the tube can be pressed. Furthermore, alternatively, the opposing unit may also be positioned at a distance from the sample cartridge and / or from the permeable planar substrate, if present within the cartridge, and the tube is pressed against a plane by the temperature control unit / surface.

[0062] In one embodiment of the device of the present invention, the temperature control surface or the opposing surface or both are preferably transparent within the range of 250 nm to 950 nm or a partial range thereof. In one embodiment, the temperature control surface or the opposing surface or both are within the range of 400 nm to 600 nm and / or within the range of 450 nm to 650 nm and / or within the range of 500 nm to 700 nm and / or within the range of 550 nm to 750 nm and / or within the range of 600 nm to 800 nm. In one embodiment, the temperature control unit of the device has a receiving portion for receiving the tube, and the receiving portion enables the fixing of the tube on the temperature control surface. In a preferred embodiment, the device according to the present invention further has one or several spacers located on the opposing unit, preferably on the opposing surface, or on the temperature control unit, preferably on the temperature control surface, or on both the opposing unit and the temperature control unit. Preferably, the spacer has a height represented by the following formula H S =2×T + H CS wherein H S = the height of the spacer, T = the wall thickness of the deformable transparent tube, and H CS = the height of the internal space of the tube when it has a flat non-circular cross-section. When there are several spacers, the height of each spacer is the same H S and preferably, H S is within the range of 7 μm to 2,500 μm, preferably 10 μm to 1,000 μm, more preferably 50 μm to 500 μm, and even more preferably 100 μm to 300 μm. For example, when a monodisperse suspension of particles with an average diameter of 50 μm and tubes with a wall thickness of 70 μm are used, a spacer height of about 190 μm is appropriate.

[0063] <> Alternatively, H cs can be adjusted by using an active movable spacer. By measuring H cs H sThis may be precisely controlled. This may be advantageous in scenarios where the pipe does not have acceptable precision in its wall thickness, for example, when a batch of pipes produced lacking an acceptable or desirable wall thickness tolerance is used. cs Regarding the determination, an optical readout system may be used (e.g., optical focus), and all other methods are also available (interferometer, TOF, capacitance, etc.).

[0064] In one embodiment, the device according to the present invention further comprises an optical detection means configured to detect and / or analyze the contents of the internal space of the tube using optical spectroscopy and / or imaging. Preferably, such detection and / or analysis is carried out using a beam path that travels through the central portion of the tube and through one or both of the temperature control surface or the opposing surface. As noted above, in one embodiment, the optical detection means also H cs It may be used to determine, or may be configured to do so.

[0065] In one embodiment, the optical detection means may be integrated into the opposing unit or the temperature control unit or both, or it may be provided separately.

[0066] In one embodiment, the device according to the present invention is configured to be capable of holding a plurality of sample cartridges according to the present invention. In one embodiment, the device comprises a plurality of sample cartridges according to the present invention.

[0067] (List of reference numbers) 100 sample cartridges 110 Deformable permeable tube 111 / 112 Two opposing ends of the pipe 113 Internal space of the pipe 114 / 114' / 114'' One or more walls of a pipe 114'' Single wall of the pipe 115 Center part of the pipe 116 Vertical axis of the pipe 120 Means of detention 130 Means for reversibly closing and sealing a pipe 140-mounted frame 141 / 142 First and second lateral sides of the mounting frame 143 Vertical axis of the mounted frame Two opposing longitudinal ends of the 144 / 145 mounting frame 146 / 147 are orifices located at the vertical ends 144 / 145, respectively. 148 / 148' means for reversibly closing and sealing orifices 146 / 147, respectively. 149 Space enclosed by the mounted frame 200 dispersions of particles, cells, or droplets 210 Particle or cell suspension 220 droplet emulsion Devices for 300 incubations 310 Temperature control unit 311 Temperature control surface 312 Receiving part of the tube 320 opposing units 321 Opposing surface 330 / 330' One or more spacers 340 Optical detection means

[0068] Features of the present invention disclosed herein, in the claims, and / or in the accompanying drawings may, individually or in any combination thereof, be materials for realizing the present invention in various forms.

Claims

1. A sample cartridge (100) for incubating and / or analyzing a dispersion (200) of particles, cells, or droplets, and / or for carrying out a biochemical reaction with or in such a dispersion, The sample cartridge comprises a deformable permeable tube (110), the deformable permeable tube (110) having two opposing open ends (111, 112), the two opposing open ends serving as an inlet and an outlet, respectively. The tube is adapted to receive a dispersion of particles, cells, or droplets, and within the internal space (113) of the tube, the internal space is lined by one wall (114'') or several walls (114, 114', 114'') of the tube, and the tube has a circular or oval cross-section when the internal space of the tube is receiving the dispersion, and the internal space has a flat, non-circular cross-section when the tube is pressed against a surface. The sample cartridge further comprises means (120) for retaining the particles, cells, or droplets within the deformable permeable tube, wherein the means for retaining the particles are located on one or both of the opposing open ends of the tube, and the means for retaining are a filter, grid, mesh, or sieve. The sample cartridge further comprises a mounting frame (140), the mounting frame (140) connected to the tube at the opposing open ends of the tube and holding the tube, the mounting frame (140) configured to allow the addition of a substance to the internal space of the tube through one of the opposing open ends acting as an inlet, and / or to allow the removal of a substance from the internal space of the tube through one of the opposing open ends acting as an outlet, the mounting frame (140) having first and second lateral sides (141, 142) arranged to face each other, The mounting frame is configured such that one of the sides of the mounting frame is exposed to a temperature control unit (310) having the temperature control surface, by enabling physical contact of the temperature control surface (311) with the central portion (115) of the tube through one of the opposing sides, and by enabling the application of pressure by the temperature control surface to the opposing surface (321) of the opposing unit (320) with respect to the central portion of the tube through one of the opposing sides, wherein the opposing unit (320) and the temperature control unit (310) are separate components that do not form part of the sample cartridge.

2. The sample cartridge according to claim 1, wherein the deformable permeable tube has a single wall (114'''), and the internal space (113) is lined by the single wall.

3. The sample cartridge according to any one of claims 1 to 2, further comprising means (130) for reversibly closing and sealing the deformable permeable tube at one or both of the opposing open ends (111, 112).

4. The sample cartridge according to any one of claims 1 to 3, wherein the mounting frame (140) is further configured to enable analysis of the central portion (115) of the tube by an optical detection means.

5. The sample cartridge according to any one of claims 1 to 4, wherein the central portion (115) of the tube is a portion that is closed and sealed by the means (130) for reversibly closing and sealing the deformable permeable tube.

6. The sample cartridge according to any one of claims 1 to 5, wherein the mounting frame has a longitudinal axis (143) aligned with the longitudinal axis (116) of the tube, the mounting frame comprises two opposing longitudinal ends (144, 145), each of which has orifices (146, 147) that fluidly connect to the opposing open ends (111, 112) of the tube, each serving as the inlet and outlet of the tube, and each of the orifices is sealable.

7. The sample cartridge according to claim 6, wherein the tube (110) is mounted within the mounting frame (140) such that the opposing open end of the tube is attached to or in contact with the opposing longitudinal end of the mounting frame, the mounting frame encompassing a space (149) through which the tube extends, the space being configured to allow exposure to or contact of the central portion (115) of the tube to a temperature control unit and / or to allow analysis of the central portion of the tube by optical detection means.

8. The sample cartridge according to any one of claims 1 to 7, wherein the walls (114, 114', 114'') of the tube have a thickness in the range of 1 μm to 1,000 μm, and / or the diameter of the internal space is in the range of 0.1 cm to 5 cm when it has a circular or oval cross-section, and / or the height of the internal space of the tube is in the range of 5 μm to 500 μm when it has a flat non-circular cross-section.

9. Use of a sample cartridge according to any one of claims 1 to 8 for generating and / or processing dispersions of particles, cells, or droplets.

10. The aforementioned process involves incubating the dispersion of particles, cells, or droplets. The use according to claim 9, which is one or more of the activities of carrying out a biochemical reaction with the dispersion of particles, cells, or droplets; binding one or more analytes to the particles, cells, or droplets and then removing any unbound analytes and other unbound substances from the particles, cells, or droplets; exchanging the liquid phase of the dispersion; and analyzing the dispersion of particles, cells, or droplets.

11. The aforementioned use is, The steps include filling the tube with a dispersion of particles, cells, or droplets, and optionally, one or more additional reagents. The steps include optionally arranging the particles, cells, or droplets in a single layer, The steps include carrying out a biochemical reaction in the tube and / or incubating the tube under one or more defined reaction conditions, The steps include analyzing the results of such biochemical reactions and / or such incubations, The use according to any one of claims 9 to 10, including the use described in any one of claims 9 to 10.

12. The aforementioned use is, A step of filling the tube with a dispersion of particles, cells, or droplets, wherein the dispersion includes a first liquid, The steps include removing the first liquid from the particles, cells, or droplets inside the tube, The steps include adding a second liquid containing the analyte to the tube, A step of incubating the particles, cells, or droplets in the second liquid to enable or facilitate the binding of the analyte to the particles, cells, or droplets, The steps include removing the second liquid from the particles, cells, or droplets in the tube, and optionally washing the particles, cells, or droplets to remove any unbound analytes and unbound substances from the particles, cells, or droplets, The steps include: resuspending the particles in a third liquid by adding such third liquid to the tube; The steps include optionally arranging the particles, cells, or droplets in a single layer, The steps include carrying out a biochemical reaction in the tube and / or incubating the tube under one or more defined reaction conditions, The steps include analyzing the results of such biochemical reactions and / or such incubations, Use according to any one of claims 9 to 11, including the use described in any one of claims.

13. A method for generating a droplet dispersion, the method comprising the steps of: providing a sample cartridge according to any one of claims 1 to 8; and mixing an aqueous phase and an oily liquid phase in the tube of the sample cartridge, thereby generating a dispersion.

14. A method for producing a solid or semi-solid dispersion, the method comprising the steps of: providing a sample cartridge according to any one of claims 1 to 8; mixing an aqueous phase and an oily liquid phase in a tube of the sample cartridge, and thus producing a dispersion, wherein one of the phases also contains solid particles or one or more components capable of forming a gel or solid in response to changing at least one environmental condition around the components, and if one of the phases also contains one or more components capable of forming a gel or solid, the method further comprises the step of inducing the formation of a gel or solid by changing the at least one environmental condition around the components, thereby converting the droplets into particles and producing a dispersion of particles in the tube.

15. A device (300) for incubating a dispersion of particles, cells, or droplets, and / or for carrying out a biochemical reaction with thereof, wherein the device is A sample cartridge (100) according to any one of claims 1 to 8, A temperature control unit (310) having a temperature control surface (311), wherein the temperature control unit (310) is adapted to heat and / or cool via the temperature control surface. Equipped with, The device is configured such that the tube of the sample cartridge can be brought into contact with the temperature control surface (311) using one of its lateral sides (141, 142), or is in contact with and can be pressed by or against the temperature control surface (311), so that the tube is deformed when pressed by or against the temperature control surface, and the internal space (113) of the pressed tube has a flat, non-circular cross-section.

16. The device according to claim 15, further comprising an opposing unit (320), the opposing unit (320) being positioned at a distance from the temperature control unit (310) so as to face the temperature control unit (310), the opposing unit (320) having an opposing surface (321) facing the temperature control surface (311), and the opposing unit (320) not forming part of the sample cartridge and being a separate component provided within the device (300) separately from the sample cartridge.

17. The device according to claim 16, wherein one or both of the temperature control surface or the opposing surface are permeable.

18. The device according to any one of claims 15 to 17, further comprising an optical detection means (340), wherein the optical detection means is configured to detect and / or analyze the contents of the internal space (113) of the tube using optical spectroscopy and / or imaging.

19. Use of the device according to any one of claims 15 to 18 for generating and / or processing dispersions of particles, cells, or droplets, wherein the use is accompanied by a sample cartridge according to any one of claims 1 to 8, and the use is performed as defined in any one of claims 9 to 12.

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