Medical Device For The Selective Separation Of A Biological Sample

Abstract

The present invention provides a medical device and a method for the selective separation of a biological sample of a mammal into a first portion and a second portion. It comprises a first layer comprising a first reservoir for receiving the biological sample and for retaining the first portion of the sample, and a second layer comprising a second reservoir for receiving the second portion of the sample. Between the first layer and the second layer a third layer is provided, wherein the third layer comprises a plurality of channels configured to provide a fluid communication between the first reservoir and the second reservoir. Furthermore, between the first layer and the second layer a fourth layer adjacent to the third layer is provided, wherein the fourth layer comprises and/or is configured as a separation layer. At least the third and fourth layer are configured to selectively separate the biological sample between the first reservoir and the second reservoir into the first portion and the second portion of the sample. According to the invention, the layers are to be understood to be stackable in a substantially vertical plane, forming a three-dimensional layered structure.

Claims

1. Medical device (1) for the selective separation of a biological sample of a mammal into a first portion and a second portion, comprising: a first layer (10) comprising a first reservoir (12) for receiving the biological sample and for retaining the first portion of the sample; a second layer (40) comprising a second reservoir (42) for receiving the second portion of the sample; a third layer (30) between the first layer (10) and the second layer (40), wherein the third layer (30) comprises a plurality of channels (32) configured to provide a fluid communication between the first reservoir (12) and the second reservoir (42); and a fourth layer (20) between the first layer (10) and the second layer (40) and adjacent to the third layer (30) comprising a separation layer (22), wherein at least the third and fourth layer (30; 20) are configured to selectively separate the biological sample between the first reservoir (12) and the second reservoir (42) into the first portion and the second portion of the sample.

2. Medical device (1) according to claim 1, wherein the fourth layer (20) comprises a porous structure located proximate a top surface of the first reservoir (12), wherein the porous structure comprises one of a multi-hole array, as triangular geometries or openings 22a defining a plurality of pores coinciding with the opening of the plurality channels (32) of the third layer (30) at the fourth layer (20).

3. Medical device (1) according to claim 1, wherein the biological sample is a fluid and/or semi-solid sample comprising semen comprising a biologically active component, wherein the biologically active component is separated from the first reservoir, wherein the biologically active component comprises sperm, preferably spermatozoa.

4. Medical device (1) according to claim 1, wherein the plurality of channels (32) of the third layer (30) may have an angular orientation relative to the first layer (10) and/or second layer (40) of at least 60 degrees, preferably an angular orientation perpendicular to the first layer (10) and/or second layer (40).

5. Medical device (1) according to claim 1, wherein the plurality of channels (32) of the third layer (30), is configured for directing movement of a biologically active component in the biological sample from the first reservoir (10) to the second reservoir (40).

6. Medical device (1) according to claim 5, wherein each channel (32) of the third layer (30) comprises a surface structure that promotes and/or supports motility of a biologically active component in the biological sample, and/or that selectively promotes and/or supports a biologically active motile component in the biological sample.

7. Medical device (1) according to claim 1, wherein each of the plurality of channels (32) of the third layer (30) is at least one of non-linear and/or comprises an arrangement of edges.

8. Medical device (1) according to claim 1, wherein each of the plurality of channels (32) of the third layer (30) has a width that is larger at the first layer (10) than it is at the second layer (40).

9. Medical device (1) according to claim 1, wherein each of the plurality of channels (32) of the third layer (30) define a meandering, sinuous, helical, and/or zig-zag path.

10. Medical device (1) according to claim 1, wherein the second reservoir (42) comprises at least one side, bottom, and/or top surface having a rounded, cylindrical, semicircular, conical, U-shaped, and/or polygonal shape, wherein preferably the at least one side, bottom and top surface is shaped to at least one of provide a retaining structure for the second portion of the sample, and/or to prevent convection and/or turbulence during flow of the second portion of the sample.

11. Medical device (1) according to claim 1, wherein the first reservoir (12) is formed as a channel or a plurality of channels, and wherein the channel or each of the plurality of channels is preferably non-linear and/or defines a meandering, sinuous, helical, pedigree, circular, ellipse, U-shaped, serpentine-shaped, and/or zig-zag path having no sharp edges.

12. Medical device (1) according to claim 1, wherein at least one of the first reservoir (12) and/or the second reservoir (42) is configured to minimize convection, fluid resistance, and/or turbulent flow when the biological sample flows through the device.

13. Medical device (1) according to claim 2, wherein the plurality of openings 22a are sized and shaped to minimize the occurrence of convection and/or pressure difference within and/or between at least the second and third layer (40; 30), and wherein the each of the plurality of openings 22a has a triangular geometry.

14. Medical device (1) according to claim 1, wherein the separation layer (22) further comprises porous structure comprising a plurality of openings 22a each having a pore size between 20 and 500 m, preferably between 50 and 200 m, in particular 100 m.

15. Medical device (1) according to claim 1, wherein the first reservoir (12) comprises a volume between 0.5 and 5 mL, the second reservoir (42) comprises a volume between 100 L and 1 mL, preferably 500 L, each of the plurality of channels (32) comprises a length between 2 and 15 mm, and/or wherein the fourth layer (20) comprises a length between 100 m and 1 mm, preferably between 400 m and 600 m, in particular 500 m.

16. Medical device (1) according to claim 1, wherein the first reservoir (12) and the second reservoir (42) each comprise an inlet and an outlet (A1, A2, A3, A4), wherein preferably at least one inlet and/or one outlet (A1, A2, A3, A4) can be selectively opened and/or closed, wherein preferably an inlet and/or outlet (A1, A2, A3, A4) of each reservoir (12; 42) is adapted to connect and/or receive a second medical device, preferably a third reservoir, a syringe, and/or catheter, and/or wherein an inlet and/or outlet (A1, A2, A3, A4) of each reservoir (12; 42) is configured to minimize convection, fluid resistance, and/or turbulent flow when the biological sample is removed from the second reservoir (42).

17. Medical device (1) according to claim 1, wherein at least one of the first reservoir (12) and/or the second reservoir (42) comprises a valve arrangement, wherein the valve arrangement preferably minimizes the occurrence of gas accumulation in the at least one of the first and/or second reservoir (12; 42).

18. Medical device (1) according to claim 1, wherein the plurality of channels (32) of the third layer (30) comprises a closing means for sealing of the third layer (30) at at least one of the first and/or second layer (10; 40), the closing means being configured to prevent convection from the plurality of channels (32) of the third layer (30) to the second reservoir (40) when a flow of the second portion of the biological sample occurs in the first or second reservoir (12; 42), and wherein the closing means preferably comprises one of a microfluidic valve, a sealable membrane, a solenoid valve, and/or diaphragm valve and is configured to be selectively closed in at least one flow direction.

19. Medical device (1) according to claim 1, wherein the first layer (10), second layer (40), third layer (30 and fourth layer (20) are unitarily formed, and wherein the device (1) is portable usable as a home-care product.

20. System for the selective separation of a biological sample of a mammal into a first portion and a second portion, comprising: a biological sample; at least one medical device (1); comprising: a first reservoir (12) for receiving the biological sample and for retaining the first portion of the sample; and a second reservoir (42) for receiving the second portion of the sample; at least one medical application device for loading the first reservoir (12) of the at least one medical device (1) with the biological sample; and a second medical application device for removing a second portion of the biological sample from the second reservoir (42) of the at least one medical device (1) and configured to administer the second portion of the biological sample to a patient.

21. System according to claim 20, further comprising a heating unit for maintaining the system at a desired temperature, preferably between 25 and 37 degrees Celsius.

22. Method for the selective separation of a biological sample of a mammal with a medical device (1) comprising the steps of: providing a medical device (a) comprising a first layer (10) comprising a first reservoir (12) for receiving the biological sample and for retaining the first portion of the sample, a second layer (40) comprising a fluidly sealed second reservoir (42) for receiving the second portion of the sample, a third layer (30) between the first layer (10) and the second layer (40), wherein the third layer (30) comprises a plurality of channels (32) configured to provide a fluid communication between the first reservoir (12) and the second reservoir (42), and a fourth layer (20) between the first layer (10) and the second layer (40) and adjacent to the third layer (30) comprising a separation layer (22), wherein at least the third and fourth layer (30; 20) are configured to selectively separate the biological sample between the first reservoir (12) and the second reservoir (42) into the first portion and the second portion of the sample; preparing at least the second reservoir (42), the separation layer (22), and the plurality of channels (32) of the third layer (30) with a buffer; loading the first reservoir (12) with a biological sample of a mammal (200); fluidly sealing (300) the first reservoir (12) without establishing a pressure gradient between the first reservoir (12) and the second reservoir (42); and selectively separating (400) the biological sample between the first reservoir (12) and the second reservoir (42) into a first portion and a second portion.

23. Method according to claim 22, wherein the second reservoir (42) comprises a surface-bound compound for selectively directing a second portion of the biological sample to the second reservoir (42), and wherein the compound preferably comprises a compound for biological molecular signaling, preferably a chemokine.

24. Method according to claim 22, further comprising the step of adjusting a plane of the medical device (1) to prevent the accumulation of gas in at least the first reservoir (12) during the loading of the first reservoir (12) with the biological sample.

25. Method according to claim 22, wherein the step of selectively separating of the biological sample occurs over a period of time ranging from 10 minutes to 60 minutes, preferably ranging from 20 minutes to 45 minutes, in particular 30 minutes, and/or wherein the medical device (1) is heated to a temperature ranging from 25 degrees to 37 degrees Celsius prior to the loading of the first reservoir (12) with a biological sample of a mammal and during the selectively separating of the biological sample.

26. Medical device (1) according to claim 1, wherein the fourth layer (20) comprises a porous structure located proximate the plurality of channels (32) of the third layer (30), wherein the porous structure comprises one of a multi-hole array, as triangular geometries or openings 22a defining a plurality of pores coinciding with the opening of the plurality channels (32) of the third layer (30) at the fourth layer (20).

27. Medical device (1) according to claim 17, wherein the valve arrangement further comprises a flow constriction means to control a flow of the biological sample with the reservoir (12; 42).

28. Method according to claim 22, wherein the preparing step further comprises one of loading (100), flushing, incubating, and/or mixing at least the second reservoir (42), the separation layer (22), and the plurality of channels (32) of the third layer (30) with a buffer

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0071] The present disclosure will be more readily appreciated by reference to the following detailed description when being considered in connection with the accompanying drawings in which:

[0072] FIG. 1A is a schematic cross sectional view of a medical device for selectively separating a biological sample of a mammal, wherein the fourth layer is adjacent to the first layer;

[0073] FIG. 1B is a schematic cross sectional view of a medical device for selectively separating a biological sample of a mammal, wherein the fourth layer is adjacent to the second layer;

[0074] FIG. 2A is a schematic top view of another medical device for selectively separating a biological sample of a mammal at different transversal sections;

[0075] FIG. 2B is a schematic perspective view of said medical device for selectively separating a biological sample of a mammal;

[0076] FIG. 2C is a schematic cross sectional view of said medical device for selectively separating a biological sample of a mammal;

[0077] FIG. 2D is a enlarged top/bottom view of the fourth layer, wherein the separation layer comprises a multi-hole or pore structure;

[0078] FIG. 3A-C show schematic cross sectional views of a medical device for selectively separating a biological sample of a mammal with alternative configurations of the plurality of channels and the second reservoir;

[0079] FIG. 4A-C show schematic perspective views a medical device for selectively separating a biological sample of a mammal with alternative configurations of inlets and outlets of the first and second reservoir;

[0080] FIG. 5 is a schematic diagram of a method for selectively separating a biological sample of a mammal

[0081] FIG. 6 shows the schematic cross sectional view of FIG. 3A to indicate a method according to the invention; and

[0082] FIG. 7 schematically shows the separation of a second portion of the biological sample from the first reservoir into the second reservoir.

[0083] FIG. 8A shows a schematic perspective view of a medical device for selectively separating a biological sample of a mammal as seen from one side of the medical device, wherein the separation layer comprises openings with triangular geometries;

[0084] FIG. 8B shows a schematic perspective view of said medical device for selectively separating a biological sample of a mammal as seen from the bottom of said medical device, the bottom wall of the first layer being removed;

[0085] FIG. 8C shows a blowup schematic bottom view of said medical device for selectively separating a biological sample of a mammal, the bottom wall of the first layer being removed;

[0086] FIG. 8D shows another schematic perspective view of said medical device for selectively separating a biological sample of a mammal as seen from the bottom of said medical device, the bottom wall of the first layer being removed;

[0087] FIG. 8E shows a schematic perspective view of said medical device for selectively separating a biological sample of a mammal, wherein the second and third layer are removed;

[0088] FIG. 8F shows a schematic perspective view of said medical device for selectively separating a biological sample of a mammal at a cross section, wherein the first and second layer are removed.

[0089] In the following, the invention will be explained in more detail with reference to the accompanying Figures. In the Figures, like elements are denoted by identical reference numerals and repeated description thereof may be omitted in order to avoid redundancies.

DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS

[0090] In FIG. 1A, a medical device 1 is depicted for the selective separation of a biological sample of a mammal into a first portion and a second portion. The medical device 1 generally comprises four layers. The bottom layer 10 comprises a first reservoir 12 for receiving the biological sample. As depicted in FIG. 1, the first reservoir 12 may comprise the majority of the first layer 10 and furthermore is in fluid communication with the fourth layer 20 as indicated by the dashed line. However, other configurations, e.g. a larger first layer relative to the first reservoir, may be provided for e.g. stability and handling purposes.

[0091] The fourth layer 20 comprises a separation layer 22, which preferably comprises a porous structure, i.e. allowing compounds with a certain size or molecular mass to pass through the porous structure to the third layer 30. For example, the fourth layer may be a cellulose-based membrane, comprising pores which are large enough for spermatozoa to pass the membrane yet are small enough to prevent agglomerates or particles to enter the third layer 30.

[0092] The third layer 30 comprises a plurality of channels 32 as depicted by the plurality of vertical through lines. The channels 32 as schematically indicated here are not drawn to scale and may hence form a bottom surface area that corresponds to e.g. the size or pore sizes of the separation layer 22.

[0093] The plurality of channels 32 of the third layer 30 are furthermore in fluid communication with the second reservoir 42 of the second layer 40. Accordingly, a fluid communication is established between the first reservoir 12 of the first layer 10 and the second reservoir 42 of the second layer 40.

[0094] FIG. 1B shows an alternative configuration of the medical device 1 depicted in FIG. 1A. Here, the fourth layer 20 is disposed between the third layer 30 and the second layer 40, i.e. the separation layer 22 being adjacent to the second reservoir 42 and the plurality of channels 32.

[0095] FIGS. 2A, 2B, and 2C depict another medical device 1, wherein the plurality of channels 32 of the third layer 30 is wider towards the first layer 10, i.e. at the fourth layer 20, as depicted. Since the fourth layer 20 may also be disposed between the third layer 30 and the second layer 40, the width of the plurality of channels may be understood as to be wider at the first layer. The narrowing of the channels 32 is depicted in FIG. 2A when comparing the top side 32b of the third layer 30 with the bottom side 32a of the third layer 30. Furthermore, the plurality of channels 32 comprises a zig-zag structure comprising an arrangement of edges, which at least has the advantage that e.g. the upwards wall-swimming behavior of motile spermatozoa is supported and/or direct towards the second reservoir 42.

[0096] The first reservoir 12 is furthermore depicted as a serpentine-shaped channel. Such a shape may e.g. facilitate the replacement of the buffer and/or the loading since the occurrence of bubble formation and/or entrapment or blocking of the first reservoir 12 by accumulating gas is prevented. Furthermore, such a shape may provide a homogenous spreading of the biological sample.

[0097] By the same token, the second reservoir 42 is depicted as a U-shaped channel. Such a shape may e.g. facilitate the removal of the second portion of the biological sample, reducing the build-up of a negative pressure, ensuring a removal of the entire portion from the second reservoir 42, and providing a laminar flow, thereby reducing convection and/or mixing of the content of the second reservoir 42 with the plurality of channels 32.

[0098] Furthermore, the medical device 1 comprises an inlet and outlet A1 and A2, and A3 and A4 for the first layer 10 and the second layer 40, respectively. The connections facilitate the loading of the biological sample and the removal of the second portion of the biological sample. The inlets and outlets A1, A2, A3, A4 are depicted to be configured as luer connectors for both handling purposes and to optimize flow characteristics, e.g. providing a laminar flow and/or reducing the occurrence of vortices, see e.g. FIG. 2B.

[0099] The narrowing of the plurality of channels 32 is furthermore depicted in FIG. 2C, forming a conical or pyramid shape. In addition, the U-shaped channel of the second reservoir 42 is depicted as a polygonal or pyramid shaped channel in a Z-plane, i.e. forming a roof-shaped structure from the bottom of the second reservoir 42. Such a shape may furthermore increase the accumulation of spermatozoa in the top of the second reservoir 42, forming a trapping system, thereby preventing highly motile spermatozoa to swim back into the plurality of channels 32, also during removal of the second portion from the second reservoir 42.

[0100] The separation layer 22 preferably comprises a porous structure or multi-hole array 24 as depicted in FIG. 2D. The holes may be linear so that the top side and bottom side are symmetrical. The indicated holes are merely indicative. Accordingly, different spacing between the holes or pores may be provided and the sizes may be varied. Alternatively, the separation layer 22 may provide a continuous porous structure or multi-hole array 24 on its entire surface or at least a substantial part thereof. However, as indicated in FIG. 2D, a porous structure or multi-hole array 24 may be configured to be aligned with the plurality of channels 32 of the third layer 30 for e.g. manufacturing purposes and/or to reduce the occurrence of misalignment. Accordingly, a mask may be used to ensure optimal alignment and prevent an impaired fluid communication of the pores or holes.

[0101] The plurality of channels 32 as well as the second reservoir 42 may have alternative configurations, as depicted in FIGS. 3A-C. The first layer 10, the third layer 30 and the fourth layer 20 are here made of a single part and form an integral part with the second layer 40. As shown in FIG. 3A, the second reservoir 42 may comprise a rounded configuration, i.e. comprising no sharp edges, at the top end of the plurality of channels 32 (compare FIG. 2A, 32b). The second reservoir 42, however, may generally comprise a polygonal shape, e.g. a pyramid shape as shown in FIGS. 3A-C, such that a corner is formed at the top of the second reservoir 42. Accordingly, highly motile spermatozoa comprising natural boundary-following swimming behavior may migrate from the first reservoir 12 through the separation layer 22 and the plurality of channels 32 to the second reservoir 42, wherein the corner at the top of the second reservoir 42 retains the spermatozoa at the top, i.e. prevents or at least impairs the spermatozoa from swimming back into the plurality of channels 32. The plurality of channels 32 is here shown to comprise a narrowing configuration along a vertical axis, i.e. being broader at the separation layer 22 than at the second reservoir 42. Again, such a configuration may facilitate both the migration of spermatozoa towards the second reservoir 42 and the retention of spermatozoa in the second reservoir 42.

[0102] Alternatively, as shown in FIG. 3B, the plurality of channels 32 may be linear, i.e. the walls not being congruent from the bottom to the top of the plurality of channels 32. Accordingly, the second reservoir 42 may comprise a larger cross sectional area, which may at least have the advantage that e.g. larger volumes may be obtained in the second reservoir 42. Here, by means of example, no rounded shape of the second reservoir 42 is indicated at the plurality of channels 32.

[0103] The second reservoir 42 may also align with more than 2 channels of the plurality of channels 32, as depicted in FIG. 3C. Accordingly, even larger volumes of the second reservoir 42 may be obtained. Such configuration may e.g. be desirable for manufacturing purposes, wherein e.g. an inlet and outlet A3, A4, are provided at either end. Accordingly, removal of spermatozoa may be facilitated. It will be understood that the described configurations only depict examples and do not limit the scope of the invention. Accordingly, other configurations and/or combinations may be provided.

[0104] As shown in FIGS. 2B and 2C, the inlets and outlets A1, A2, A3, A4 may extend from the sides of the first layer 10 and the second layer 40. However, other configurations may be implemented, as depicted in FIGS. 4A-C. For example, as shown in FIG. 4B, the inlet and outlet A3, A4 of the second layer 40 may extend from the top of the second reservoir 42 and the second layer 40, wherein e.g. hydrostatic pressure may be used to facilitate removal of the second portion. In addition, the inlet and outlet A1, A2 of the first layer 10 may extend from only one side. This configuration at least has the advantage that the medical device 1 can be held more easily by a user, i.e. having solid and/or homogeneous sides without technical features at 4 sides. This furthermore reduces the risk of contamination. FIG. 4C shows another alternative, wherein the inlets, e.g. A1 and A3, are disposed above each other and the outlets, e.g. A2 and A4, are disposed above each other. This may reduce confusion and/or increases user-friendliness by having one side for injection or loading and one side for removal or aspiration. Furthermore, this has at least the advantage that the medical device 1 is smaller in a vertical dimension, may allow better scalability of the reservoirs 12, 42, and ensures that the medical device 1 is stackable.

[0105] The inlets and outlets A1, A2, A3, A4, may furthermore be configured as or comprise safety means. For example, they may be configured to provide a snap-fit, a luer connector, or a luer lock, known in the art. Accordingly, in the event of increased tension or pressure, such a safety means prevents a connected medical application device to be released inadvertently. Accordingly, the safety means may comprise a foolproof or poka-yoke configuration, e.g. by providing a hermaphroditic connector, an omnidirectional connector, or by providing a mechanical fit, which ensures a connection to be fail-safe. The connection may furthermore be interlocking and/or provide a clearance fit, transition fit, or interference fit. In addition, the safety means may comprise a regulating, blocking, and/or locking mechanism, which is activated when e.g. a desirable or predefined maximum pressure or flow is exceeded. Such a feature may e.g. be provided by valve arrangements or the like, as described above.

[0106] FIG. 5 depicts schematically the steps of a method according to the invention for the selective separation of a biological sample of a mammal with an above described medical device. In a first step 100, at least the second reservoir 42, the separation layer 22, and the plurality of channels 32 of the third layer 30 are loaded with a buffer. The buffer may be lyophilized and already within the medical device 1, such that the injection of sterile water and mixing may be sufficient. Alternatively, a pre-mixed buffer may be injected. The injection may occur with a needle and a syringe if at least one of the layers is made of a material that allows puncture with a needle such as e.g. a biocompatible elastomer. After loading, the needle and syringe are retracted, wherein the elastomer is formed such that no fluid leaks from the first reservoir 12. However, the medical device 1 preferably comprises inlets and outlets A1, A2, A3, A4, for injection. Alternatively, the medical device 1 may be provided with a buffer by submersion into a corresponding buffer volume. Furthermore, said compartments may be flushed, incubated, and/or mixed with said buffer. In addition, the second reservoir, the plurality of channels of the third layer, the separation layer, and the first reservoir may be sterilized, flushed, and/or calibrated before the loading of said compartments with the buffer, e.g. with ethanol, sterile and/or distilled water, and buffers, respectively.

[0107] The medical device 1 may then be optionally pre-heated to preferably 37 degrees Celsius in a second, optional, step 200, prior to loading of the biological sample into the first reservoir 12. Alternatively, the medical device 1 may be kept at room temperature, e.g. between 20 and 25 degrees Celsius. However, to facilitate e.g. liquefaction of seminal plasma and for biocompatibility reasons, 37 degrees Celsius may be preferable. The buffer and/or medical device 1 may optionally also be pre-heated prior to the loading of the respective layers (10; 20; 30; 40) of the medical device 1 with buffer, step 110. Accordingly, the medical device 1 may also be retained at room temperature, i.e. not heated, before and/or after loading of the medical device 1 with buffer.

[0108] By the same token, the biological sample may be pre-heated and/or incubated to preferably 37 degrees Celsius prior to loading of the first reservoir 12. This at least has the advantage that a liquefaction of e.g. semen may occur, the handling of the biological sample may be facilitated, and/or may provide better biocompatibility or viability of the biological sample. For example, a pre-incubation of the biological sample at 37 degrees Celsius between 1 and 60 minutes, preferably between 10 and 30 minutes, may be preferable.

[0109] The first reservoir is then loaded with a biological sample of a mammal in a third step 300, wherein the second reservoir 42 is fluidly sealed. The loading of the first reservoir 12 with the biological sample may, if present, replace a buffer in the first reservoir 12. Such loading may e.g. occur with a syringe and a corresponding inlet or outlet A1, A2 of the first reservoir 12 by e.g. injection of a fluid or air. However, the first reservoir 12 and the first layer 10 may be made of a material that allows puncture with a needle such as e.g. a biocompatible elastomer. Accordingly, the loading may alternatively occur with a needle and a syringe, which may be preferable for handling or hygiene purposes. After loading, the needle and syringe are retracted, wherein the elastomer is formed such that no fluid leaks from the first reservoir 12. However, for safety reasons, the use of sharp needles may be not preferable. Furthermore, needles may only be used after sufficient liquefaction of seminal plasma for handling purposes. Appropriate needle dimensions may hence be provided to reduce safety issues, facilitate handling, and/or reduce adverse effects on the quality and/or DNA integrity of spermatozoa.

[0110] The first reservoir is accordingly fluidly sealed in a fourth step 400, without establishing a pressure gradient between the first reservoir 12 and the second reservoir 42. The prevention of a pressure gradient and/or convection may e.g. be accomplished by the configuration of the plurality of channels 32 and/or the configuration of the separation layer 22.

[0111] The biological sample is then selectively separated between the first reservoir 12 and the second reservoir 42 into a first portion and a second portion in a fifth step 500. The selective separation preferably occurs at 37 degrees Celsius, e.g. by heating, but may alternatively occur at room temperature. Furthermore, the selective separation preferably occurs due to a combination of directional motility and gravitational force. For example, spermatozoa may be selectively separated from non-motile sperm due to their natural wall swimming character. Preferably, the plurality of channels 32 of the third layer 30 are thereby substantially oriented towards or aligned with a plane of gravitational force. This prevents or at least reduces the undesirable accumulation of non-motile sperm cells, sediment, and particles into the plurality of channels 32 of the third layer 30. This not only prevents the occurrence of a potential block, impairing movement of highly motile spermatozoa towards the second reservoir 42, but also increases purity and enrichment of motile spermatozoa with high quality DNA at the top of the second reservoir 42.

[0112] FIG. 6 shows an example of the different stages of the method according to the invention. By means of example, the medical device 1 is depicted as a cross sectional side view similar to the embodiment shown in FIG. 3A. As indicated in step I.i, the first reservoir 12, the second reservoir 42, the plurality of channels 32 and separation layer 22 therebetween, may be filled with a buffer (grey). Alternatively, the first reservoir 12 may remain empty, as indicated by step I. ii (white). Both steps hence resemble the method step 100, as depicted in e.g. FIG. 5.

[0113] Step II. depicts how the first reservoir 12 of the medical device 1 may then be filled with a biological sample (black), similar to the method step 300 shown in FIG. 5. The biological sample (black) may hence either replace the buffer (grey) or fill the first reservoir, according to step I.i and I.ii, respectively. Alternatively, the first reservoir 12 of may be filled with the biological sample before the filling of the second reservoir 42, plurality of channels 32, and the separation layer 22, i.e. step I.ii would follow after filling the first reservoir 12 with the biological sample. After fluidly sealing all compartments, the separation of the second portion into the second reservoir 42 may then take place (not shown).

[0114] Step III. shows how a second portion (not shown) may be removed from the second reservoir 42 after the separation of the second portion from the biological sample into the second reservoir 42. As indicated, the second portion is removed together with the buffer, leaving the second reservoir 42 empty (white) or at least removing a substantial volume from the second reservoir 42. As described above, the removal may occur according to various alternatives, e.g. by injecting air or injecting a fluid at an inlet and accordingly receiving the second portion in the buffer at the outlet of the second reservoir 42. Alternatively, the second portion may be obtained by e.g. aspiration or suction.

[0115] The separation of the second portion of the biological sample between the first reservoir 12 and the second reservoir 42 is schematically depicted in FIG. 7, according to, by means of example, the medical device 1 of FIG. 3A. After loading of the first reservoir 12 at an inlet (A1, A2, A3, A4), depicted in I., the biological sample is provided in the first reservoir 12. The biological sample is here depicted as a mixture of circles (empty) and dots (black) representing a first portion and second portion, respectively. Accordingly, the second portion may e.g. comprise high quality spermatozoa while the first portion may e.g. comprise other cells, dead sperm cells and/or seminal plasma.

[0116] After distribution, the circles, e.g. dead cells, settle at the bottom of the reservoir 12 due to e.g. gravitational force and/or random motion. However, the dots, e.g. high quality spermatozoa, migrate towards the second reservoir 42 due to a combination of their natural boundary-following and upward swimming behavior. Accordingly, the dots pass through the separation layer 22 and enter the plurality of channels 32, preferring to follow edges and/or walls in contrast to random swimming behavior of e.g. low quality spermatozoa. The narrowing of the plurality of channels 32 at the top, i.e. at the second reservoir 42, as shown in FIG. 7, not only directs spermatozoa but furthermore prevents or at least impairs a back-swimming after the dots reach the second reservoir.

[0117] The configuration of the second reservoir 42 at the top is chosen to retain the majority of high quality spermatozoa at the top of the reservoir 42. For example, the provision of edges, as depicted in e.g. FIG. 7, facilitates the accumulation of high quality spermatozoa. Towards the end of step II., the second portion, e.g. spermatozoa, may hence be retrieved from the second reservoir 42, e.g. at an outlet of the second reservoir 42, as depicted in FIG. 7. Accordingly, the medical device 1 allows for an efficient separation of the second portion of the biological sample between the first reservoir 12 and the second reservoir 42 due to a combination of the configuration of the medical device 1, biological activity of the second portion, e.g. natural boundary-following and upward swimming behavior, and/or gravity.

[0118] The separation layer 22 of the fourth layer 20 may comprise alternative configurations. Accordingly, instead of or as a configuration of e.g. a porous structure or multi-hole array, said layer may comprise triangular geometries as depicted in FIGS. 8A-F. FIG. 8A shows a side view of such a configuration, wherein the separation layer 22 is interposed between the first layer 10 and the third layer 30. Although both the third layer 30 and fourth layer 20 are depicted to comprise an outer zig-zag structure, other configurations such as a continuous, smooth and/or homogenous outer wall may be provided in alternative embodiments. However, a zig-zag structure may at least have the advantage that a better grip or holding of the medical device 1 may be provided while simultaneously facilitating a stronger structural support.

[0119] The triangular geometries provide openings 22a that establish a fluid communication between the reservoir 12 of the first layer 10 and the plurality of channels 32 of the third layer 30, as generally depicted in FIGS. 8B-8F. The openings 22a are shown to be linear so that the top side and bottom side are symmetrical. However, alternative configurations, e.g. wherein the openings 22a are congruent from the first layer 10 towards the third layer 30, may be provided. Preferably, at least one corner of each opening 22a is an extension of the triangular wall structure in the plurality of channels, such that the openings 22a are aligned, preferably fully aligned with the plurality of channels 32 of the third layer 30, as best depicted in e.g. FIGS. 8D and 8F. The size and the number of the openings 22a as well as the spacing between said openings are merely indicative, such that other patterns including e.g. smaller and/or more triangles may be provided. Whereas FIGS. 8E and 8F depict the openings 22a of the separation layer 22 as an extracted view, FIGS. 8B-8D depict said openings in a solid depiction of the separation layer 22.

[0120] The at least partial alignment of the openings 22a with the plurality of channels 32 of the third layer 30 has at least the advantage that manufacturing of the device is facilitated and is possible using e.g. 3D printing with even low density printers. This allows for cheap prototyping and manufacturing and doesn't require any assembly steps since the device may be printed in one piece. Also, by providing corners that sperm can swim in, the triangular opening provide immediate guidance for sperm after leaving the first reservoir. Accordingly, the separation layer 22 may facilitate the selective separation of a biological sample of a mammal, in particular of sperm.

[0121] Although the size and geometry of the openings 22a may vary, said openings 22a preferably comprise dimensions between 0.5 mm0.5 mm0.5 mm and 5 mm5 mm5 mm, preferably between 1 mm1 mm1 mm and 3 mm3 mm3 mm, more preferably 1.4 mm1.4 mm2 mm, while comprising a height substantially corresponding to a height of the separation layer 22 in the range of 0.5 to 5 mm, preferably between 1 and 3 mm, more preferably 2 mm.

[0122] The embodiment as depicted in FIGS. 8A-F furthermore comprises a first layer 10, wherein the corners of said layer comprise a rounded shape. Accordingly, said shape may substantially correspond to the shape of the first reservoir 12. The rounded shape at least has the advantage that flow performance of the biological sample is improved when loaded into said first reservoir 12 through one of the inlets A1, A2. Furthermore, the rounded corners may increase the usability or handling of the medical device 1 by a user. However, other shapes including rectangular and/or solid shapes may be implemented such that the outer wall of the first layer 10 may enclose or receive the first reservoir 12, e.g. the bottom of said reservoir.

[0123] By the same token, the plurality of channels 32 of the third layer 30 are depicted in FIGS. 8A-F to not comprise a congruent, narrowing, or pyramid shape, i.e. not comprising a shape that is wider at the fourth layer 20 than at the second layer 40. However, such configurations as well as other configurations, as described in the above, may also be provided.

[0124] Furthermore, the second layer 40 is depicted comprising the second reservoir 42 with a roof-shaped configuration. This at least has the advantage that e.g. sperm may follow the boundaries and corners provided by said roof-shaped configuration and is hence retained or at least substantially prevented from leaving the second reservoir 42 after entering said reservoir. However, other configurations of the second reservoir 42, as described in the above, may be provided.

[0125] The orientation of the inlets and outlets A1, A2, A3, A4 is depicted in FIGS. 8A-D such that the inlet and outlet A1, A2 of the first layer 10 or reservoir 12 are oriented in opposite directions in one orientation plane, whereas the inlet and outlet A3, A4 of the second layer 40 or reservoir 42 are oriented in the same direction, i.e. are orientated in parallel in another orientation plane. All inlets and outlets are furthermore arranged at one end or side of the medical device 1. This configuration at least has the advantage that handling of the medical device 1 may be facilitated, leaving enough space to hold the medical device 1, and furthermore reduces the risk of confusion for a user by performing different steps, e.g. the loading of the first reservoir 12 and the removal of a second portion from the second reservoir 42, in different planes of orientation. However, other orientations of said inlets and outlets may be provided, as described above and/or depicted in e.g. FIGS. 4A-C.

[0126] It will be obvious for a person skilled in the art that these embodiments and items only depict examples of a plurality of possibilities. Hence, the embodiments shown here should not be understood to form a limitation of these features and configurations. Any possible combination and configuration of the described features can be chosen according to the scope of the invention.

LIST OF REFERENCE NUMERALS

[0127] 1 Medical device [0128] 10 First layer [0129] 12 First reservoir [0130] 20 Fourth layer [0131] 22 Separation layer [0132] 22a Openings [0133] 24 Multi-hole array or porous structure [0134] 30 Third layer [0135] 32 Plurality of channels [0136] 32a Bottom of third layer [0137] 32b Top of third layer [0138] 40 Second layer [0139] 42 Second reservoir [0140] 100 First method step [0141] 200 Second method step [0142] 300 Third method step [0143] 400 Fourth method step [0144] 500 Fifth method step [0145] A1 Inlet or outlet of first layer/reservoir [0146] A2 Inlet or outlet of first layer/reservoir [0147] A3 Inlet or outlet of second layer/reservoir [0148] A4 Inlet or outlet of second layer/reservoir