Fluid conduit holder and associated method

Abstract

A fluid conduit holder (1) comprising a support (10) and a guide (20) wherein the support (10) is configured, in use, to receive and position a fluid conduit (100) relative to the guide (20) such that the guide (20) provides a tapered path to a lumen (110) of the fluid conduit (100).

Description

Technical Field

[0001] The present invention relates to a fluid conduit holder. The fluid conduit holder is used to accommodate a fluid conduit, such as a micropipette, to assist in delivering a fluid, such as an operating liquid (such as the operating liquid disclosed in WO2021094588) and / or sperm, to the fluid conduit, which can be used, for example, in piezo-mediated microinjection or piezo-mediated intracytoplasmic sperm injection (Piezo-ICSI).

[0002] The present invention also relates to a method for filling a fluid conduit using the fluid conduit holder, for example, for in vitro fertilization of oocytes by Piezo-ICSI. Background Art

[0003] Male factor infertility is the cause of approximately 50% of couples being unable to conceive. In the 1980s, many surgeries were developed to address fertilization failure due to male gamete dysfunction, including injecting a single sperm cell (sperm) under the zona pellucida of the oocyte, but the success rate was very limited. However, it was during the performance of subzonal injection of the oocyte that the egg membrane was accidentally breached and the sperm was delivered into the ooplasm, subsequently laying the foundation for the development of intracytoplasmic sperm injection, which is still used in humans today.

[0004] Intracytoplasmic sperm injection (ICSI) is an in vitro fertilization procedure in which a sperm cell is directly injected into the cytoplasm of an oocyte. ICSI is performed by a human operator in a culture dish under a microscope using a micromanipulator and a micropipette to handle the oocyte and sperm. A fine glass micropipette (holding pipette) stabilizes the mature oocyte by gentle suction applied by a microsyringe, while a single sperm is collected from the other side using a thin and sharp glass micropipette (injection micropipette), and the sperm is immobilized by damaging the tail of the sperm using the tip of the micropipette. The mechanical force applied by the operator through the micromanipulator is used to pierce the zona pellucida and plasma membrane (egg membrane) of the oocyte using the injection micropipette. Then the sperm is injected into the cytoplasm of the oocyte. ICSI has helped more than two million babies be born globally. However, ICSI is a labor-intensive technique that requires highly skilled operators and still has room for improvement in terms of technical standardization and success rate.

[0005] Since conventional ICSI cannot solve the fertilization-related problems in mouse in vitro fertilization, an alternative oocyte microinjection method called "piezoelectric-mediated" ICSI (Piezo-ICSI) was developed for mouse oocytes in 1995. Piezo-ICSI is similar to conventional ICSI (c-ICSI) in that a micropipette containing a single sperm pierces the oocyte to deliver the sperm into the oocyte cytoplasm. However, in c-ICSI, the zona pellucida and the oolemma are pierced by mechanical force applied by the operator to the micropipette through a micromanipulator, while in Piezo-ICSI, the piezoelectric effect is utilized to pierce the oocyte. The piezoelectric effect is the phenomenon in which certain materials accumulate electric charge in response to mechanical stress, and conversely, mechanical stress can be induced by applying an electric power. The piezoelectric effect has many technical applications. In the case of piezo-ICSI, a short "piezoelectric pulse" is applied to the micropipette containing the sperm to generate an ultra-fast, sub-micron forward momentum of the micropipette. This precise and rapid movement is used to pierce the zona pellucida and the oolemma in a way that exerts less pressure on the oocyte than the mechanical force used in c-ICSI.

[0006] However, both ICSI and piezo-ICSI remain labor-intensive techniques that require highly skilled operators, and the present invention aims to improve this. Summary of the Invention

[0007] The present invention provides a fluid conduit holder that includes a support member and a guide member, wherein the support member is configured to receive a fluid conduit in use and position the fluid conduit relative to the guide member such that the guide member provides a tapered path to the lumen of the fluid conduit.

[0008] The present invention also provides a method of filling a fluid conduit using a fluid conduit holder that includes a support member and a guide member, the method comprising:

[0009] i. positioning the fluid conduit relative to the guide member such that the guide member provides a tapered path to the lumen of the fluid conduit; and

[0010] ii. at least partially filling the fluid conduit with a fluid via the guide member.

[0011] The present invention also provides additional embodiments as claimed in the dependent claims.

[0012] The claimed invention generally provides a subsystem that is suitable for use in a fluid delivery system or is configured for use in a fluid delivery system. The subsystem can generally form part of a fluid delivery system, and specifically, can form part of a fluid delivery system for in vitro fertilization.

[0013] The claimed invention advantageously assists an operator in delivering a fluid, which may include sperm and / or an operating fluid for a particular application, into a fluid conduit, such as a micropipette or a microcapillary pipette, which may be made of glass and may be used, for example, in piezo-mediated microinjection or piezo-mediated intracytoplasmic sperm injection (Piezo-ICSI), which may require precision and effective control. The operating fluid is typically back-filled. Sperm or other cells are typically in a holding medium, and the sperm holding medium is typically inserted from the front. The claimed invention can assist in filling from either one or both ends of the fluid conduit. Thus, the claimed invention can shorten the preparation time and reduce the risk and likelihood of accidents, thereby reducing waste and increasing the success rate.

[0014] The fluid conduit can be transported in a holder, thereby reducing the risk of damage to the conduit, which may be made of glass and thus fragile, and / or making the conduit easy to remove for attachment to a micromanipulator. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Embodiments of the present disclosure will now be described, by way of example only, with reference to the accompanying drawings, in which:

[0016] Figure 1 is a schematic perspective view of a first fluid conduit holder according to some embodiments of the present disclosure.

[0017] Figure 2 is a schematic perspective view of a package for a fluid conduit holder according to some embodiments of the present disclosure.

[0018] Figure 3a and Figure 3b is a schematic cross-sectional view of an interface between a fluid conduit holder and a fluid conduit according to some embodiments of the present disclosure.

[0019] Figure 4 is a schematic perspective view of a second fluid conduit holder according to some embodiments of the present disclosure.

[0020] Figure 5a and Figure 5b is a picture of a second fluid conduit holder according to some embodiments of the present disclosure.

[0021] Figure 6a and Figure 6b is a picture of a third fluid conduit holder according to some embodiments of the present disclosure. DETAILED DESCRIPTION

[0022] Aspects and features of certain examples and embodiments are described herein. Some aspects and features of certain examples and embodiments may be implemented routinely and, for the sake of brevity, these aspects and features are not described in detail. Accordingly, it will be understood that aspects and features of the devices and methods discussed herein that are not described in detail may be implemented according to any suitable routine techniques.

[0023] Figure 1 is a schematic perspective view of a first fluid conduit holder 1 according to some embodiments of the present disclosure. Figure 1 The fluid conduit holder 1 is shown, which includes a guide 20 and three axially aligned rectangular block supports 10a, 10b, 10c for receiving and supporting a fluid conduit 100. Although the fluid conduit 100 itself is not critical to the present invention, the fluid conduit 100 may include a needle, a syringe, or a pipette, specifically a micropipette. The support 10 includes a first support 10a remote from the guide 20, plus a second support 10b and a third support 10c, wherein the guide 20 is located on the third support 10c, and the second support 10b is located between the first and third supports as an intermediate support. In Figure 1 the example of, each of the three supports 10 includes an axial recess (not shown, such as a channel for receiving a portion of the fluid conduit 100) and is fixed to a base 30 of a protective sealed package for the holder 1, which will be described later with reference to Figure 2 for description.

[0024] Figure 1 The guide in includes a funnel 20 adjacent to the third support 10c. When the holder 1 receives the fluid conduit 100 in or on the support 10 during use, the support 10 will position the fluid conduit 100 relative to the guide 20 such that the guide 20 provides a tapered (narrowing) path to the lumen 110 of the fluid conduit 100. The guide 20 is configured to guide a user attempting to insert a container containing a fluid to be pre-filled into the lumen 110 of the fluid conduit 100 such that the container can be more easily inserted into the lumen 110 and the user can fill the fluid conduit 100 with the fluid from the container.

[0025] The guide 20 includes a tapered guide to provide a narrowing path to the lumen 110 of the fluid conduit 100. Preferably, the guide 20 is configured to provide a tapered path that guides towards the lumen 110 on at least two perpendicular axes. In Figure 1In it, the funnel guide 20 tapers towards the inner cavity 110 along two vertical axes (i.e., the y-axis and the z-axis perpendicular to the x-axis along which the fluid conduit 100 extends), and thus provides a path to the inner cavity 110 that is guided (or restricted from moving away) towards the inner cavity 110 in four Cartesian degrees of freedom (i.e., + / −y and + / −z). In other words, an object in the funnel 20 (such as a container to be inserted into the inner cavity of the fluid conduit 100) is restricted in movement within the funnel 20 in four Cartesian degrees of freedom and is thus guided to the inner cavity 110.

[0026] Figure 1 The funnel guide 20 in it is also configured to be axially aligned with the fluid conduit 100 when the conduit 100 is received in the holder 1, and the inner cavity of the funnel guide 20 is substantially concentric with the inner cavity 110 of the fluid conduit 100. In embodiments where the support 10 includes a recess for receiving the fluid conduit 100, the inner cavity of the guide 20 can be substantially concentric with the inner cavity of the recess to provide proper alignment when the fluid conduit 100 is received in the recess.

[0027] In Figure 1 an example, the guide includes a funnel 20 that advantageously tapers along two vertical axes and guides (restricts) movement in four Cartesian directions (x, y, z) degrees of freedom (only allowing free movement in the axial direction through the funnel 20 and restricting movement away from the inner cavity 110 when approaching the inner cavity 110). More generally, the guide 20 can include a funnel or cone structure, or include conical, frustoconical, concave, convex, or tapered surfaces, such as a part of a funnel or cone structure (e.g., a half-funnel). In some embodiments, the guide 20 provides a tapered path that guides towards the inner cavity 110 along at least two vertical axes, and / or provides a path to the inner cavity 110 that is guided (or restricted from moving away) towards the inner cavity 110 in at least three Cartesian degrees of freedom.

[0028] The guide 20 can be an element independent of the support 10, as Figure 1 shown, or the guide can be integrally incorporated into one or more of the plurality of supports 10 (described later), as Figures 6a to 6bAs shown. The size of the guide 20 is appropriately designed to fit the fluid conduit 100 to be used, and this application particularly focuses on holders suitable for micro-sized fluid conduits 100 (such as P10, P20, and P50 micropipettes suitable for volumes of 0.5 μl to 10 μl, 2 μl to 20 μl, and 5 μl to 50 μl, respectively). For Piezo-ICSI, in most cases, the amount of operating liquid pre-filled into the micropipette is between 10 μl and 20 μl. In some embodiments, the fluid conduit 100 has a volume of substantially 0.5 μl to 50 μl, 2 μl to 30 μl, 5 μl to 25 μl, 10 μl to 20 μl, or 15 μl, and / or may have an inner diameter of substantially 0.5 μm to 20 μm, 1 μm to 10 μm, 2 μm to 8 μm, 3 μm to 7 μm, 4 μm to 6 μm, 3.5 μm to 5.5 μm, or 4.5 μm to 5.5 μm. For such embodiments, the guide 20 may include a narrow end, the opening size of which (such as the diameter of the funnel, the width or height of the curved surface) is substantially the same as the opening size of the inner diameter of the fluid conduit 100 during use, or may be slightly larger than the opening size of the inner diameter of the fluid conduit (to assist insertion but not overly restrict the inner diameter of the fluid conduit 100), or may be slightly smaller than the opening size of the inner diameter of the fluid conduit (possibly providing a direct path to the lumen 110 of the fluid conduit 100). In some embodiments, the narrow end opening size of the guide 20 can thus be substantially 0.5μm to 50 μm, 1 μm to 25 μm, 2 μm to 20 μm, 3 μm to 15 μm, 3 μm to 10 μm, 4 μm to 6 μm, 4.5 μm to 5.5 μm, or 3 μm to 5 μm.

[0029] During ICSI surgery, sperm holding medium is aspirated from the front end, typically about 2 μl to 10 μl. In piezo-ICSI, first, the micropipette is pre-filled with an operating liquid of typically about 15 μl, and then the operating liquid is pulled down to the front tip. After that, sperm holding medium is aspirated from the front tip such that the sperm holding medium is in direct contact with the operating liquid.

[0030] Accordingly, the present invention provides a fluid holder 1 to assist an operator in filling a fluid conduit 100, which provides a tapered path to the lumen 110 of the fluid conduit 100 in use when the fluid conduit is located in the holder 1. In use, the operator positions the fluid conduit 100 relative to the guide 20 (e.g., substantially adjacent to the guide 20, or inserting the guide 20 into the conduit 100, or inserting the conduit 100 into the guide 20), such that the guide 20 provides a tapered path to the lumen 110 of the fluid conduit 100, thereby assisting in inserting a container containing the fluid to be filled into the lumen 110 of the fluid conduit 100. In some embodiments, the guide 20 may be fluidly connected to the end of the fluid conduit 100 in use.

[0031] Figure 2 is a schematic view of a package for the fluid conduit holder 1 according to some embodiments of the present disclosure. The package includes a base 30, side walls 35, and a removable protective tear-off membrane cover 40 that covers the support 10 and the guide 20 therein. The package can provide a safe and sterile housing for the holder 1. The operator removes the membrane cover 40 to expose the holder 1 ready for use.

[0032] Figure 3a and Figure 3b is a schematic cross-sectional view of the interface between the fluid conduit holder 1 and the fluid conduit 100 according to some embodiments of the present disclosure. As Figures 3a to 3b shown, in use, in some embodiments, the fluid conduit 100 may be received in the guide 20, as Figure 3a shown; or the guide 20 may be received in the fluid conduit 100, as Figure 3b shown. The size of the guide 20 is appropriately designed for the intended fluid conduit 100 and the intended interface method. The guide 20 may be interchangeable / replaceable, and for example, multiple guides 20 of different sizes may be provided in the package. In some embodiments, the fluid conduit 100 and / or the guide 20 are keyed on suitable (inner / outer) surfaces to provide a more secure engagement mechanism. In Figure 3a , the fluid conduit 100 is received in the narrow end of the guide 20, and the guide 20 abuts around the outside of the fluid conduit 100, or alternatively, is fixed around the outside of the fluid conduit, for example, using a compressible interference fit. As Figure 3a shown, the guide 20 (alternatively, the fluid conduit 100) may include a sealing element 60, such as an O-ring, which is configured to assist in fluid sealing between the fluid conduit 100 and the guide 20. In Figure 3b , the guide 20 is received within the lumen 110 of the fluid conduit 100. As Figure 3bAs shown, the guide member 20 may include a sealing element 60, such as a compressible, impermeable material on the outer surface of the guide member, to assist in providing a fluid seal between the fluid conduit 100 and the guide member 20. Alternatively, the fluid conduit 100 may be provided with a sealing element 60 on its inner wall.

[0033] Figure 4 is a schematic perspective view of a second fluid conduit holder 1 according to some embodiments of the present disclosure. Key differences from Figure 1 the first embodiment are outlined here. In this second embodiment, the holder 1 includes only two support members 10: a first support member 10a remote from the guide member 20 for supporting the distal end of the fluid conduit 100; and a second support member 10b for supporting both the guide member 20 and the proximal end of the fluid conduit 100 in use. In this embodiment, the first support member 10a extends axially beyond the length of the fluid conduit 100 and provides a recess (see the inserted cross-sectional view) for receiving a retaining element 70 to retain the fluid conduit 100 therein. The retaining element 70 may include a flexible material and / or an elastic material that restricts movement of the fluid conduit 100 within the recess, thereby holding the fluid conduit in place. The first support member 10a extends axially beyond the end axial length of the fluid conduit 100 to assist in protecting the distal end (which may be sharp) of the fluid conduit 100 in use.

[0034] In Figure 4 the embodiment, the guide member 20 includes a concave surface 20 that tapers (narrows) towards the lumen 110 in width along the y-axis. The inserted cross-sectional view shows that the guide member 20 is concave in cross-section (perpendicular to the axis along which the fluid conduit 100 extends) and effectively acts as a semi-funnel. Figure 5a and Figure 5b are pictures of the second fluid conduit holder 1 that more clearly show the concave surface guide member 20. These images show that the guide member 20 is configured to guide in two perpendicular axes, thereby providing a path to the lumen 110 in use that is guided (restricted) in three Cartesian degrees of freedom: first (best shown in the Figure 4 inset of Figure 5a and Figure 4 by the tapering of the concave shape to guide (restrict) movement in two y-direction degrees of freedom, which concave shape narrows in width (width along the y-axis) as it approaches the lumen 110 (along the x-axis or z-axis); and second, by the axial extension of the guide member 20 along the x-axis, the guide member 20 may be horizontal as shown, for example, in Figure 5a or inclined towards the fluid conduit 100 on the z-axis (as shown in Figures 5a to 5bThe guide 20 operates effectively like a semi - funnel, which does not limit movement in the positive z - direction degree of freedom and only limits movement in the negative z - direction.

[0035] Figure 6a and Figure 6b is a picture of a third fluid conduit holder according to some embodiments of the present disclosure. In this embodiment, the holder 1 includes a first support member 10a that is integrally incorporated into the packaging base 30 and is remote from a second support member 10b that includes the guide 20, and the two support members 10 have V - shaped channel recesses. As Figure 6b best shown, the guide 20 includes a tapered recess in the support member 10 at the axial proximal end of the support member, thus effectively providing an integrally incorporated funnel. In use, with the fluid conduit 100 located in the channel recess of the support member 10, the guide 20 thus provides a path to the lumen 110 of the fluid conduit 100 that tapers on two perpendicular axes and guides towards the lumen 110 on two perpendicular axes. Thus, as Figure 1 shown, the tapered recess tapers on two perpendicular axes and thus provides a path to the lumen 110 that is guided (restricted) in four Cartesian degrees of freedom.

[0036] The holder 1 further includes an upright member 50 in the form of a wall that is integrally incorporated into the packaging base 30 and is similar to the first support member 10a. The upright wall 50 positions and holds the second support member 10b and can additionally support a protective tear - off lid 40 for the packaging of the holder 1 (shown in Figure 2 ).

[0037] In some embodiments, the guide 20 is configured to be axially aligned with the fluid conduit 100 when the conduit 100 is received in the holder 1. In other embodiments, the guide 20 further includes a flexible tube, such as a telescopic tube or a bellows (not shown), which allows the guide 20 to move out of axial alignment (and / or extend) with the fluid conduit 100 to adjust the entry angle of the guide 20, thereby further assisting the user.

[0038] The various embodiments presented herein are provided only to assist in understanding and teaching the claimed features. These embodiments are provided only as representative samples of embodiments and are not exhaustive and / or exclusive. It should be understood that the advantages, embodiments, examples, functions, features, structures, and / or other aspects described herein should not be considered as limitations on the scope of the invention defined by the claims or on equivalents of the claims, and that other embodiments may be used and modifications may be made without departing from the scope of the claimed invention.

[0039] In addition to those specifically described herein, various embodiments of the present invention may suitably include, consist of, or consist essentially of a suitable combination of the disclosed elements, components, features, parts, steps, devices, etc. Further, the present disclosure may include other inventions that are not currently claimed but may be claimed in the future. Protection may also be sought for any features disclosed in any one or more of the documents incorporated herein by reference and combined in the present disclosure.

[0040] Index of Reference Signs

[0041] Any reference signs in the claims shall have no limiting effect on the scope of the claims but are merely for assistance to the reader.

[0042]

Claims

1. A fluid conduit retainer, comprising a support member and a guide member, wherein, The support member is configured to receive a fluid conduit in use and position the fluid conduit relative to the guide member such that the guide member provides a tapered path to the lumen of the fluid conduit.

2. The retainer according to claim 1, wherein, The guide member includes a funnel or cone structure, or includes a conical, frustoconical, concave, convex, or tapered surface.

3. The retainer according to any one of the preceding claims, wherein In use, the guide member provides a tapered path to the lumen that guides in at least three Cartesian degrees of freedom.

4. The retainer according to any one of the preceding claims, wherein, In use, the guide member provides a tapered path to the lumen that guides in four Cartesian degrees of freedom.

5. The retainer according to any one of the preceding claims, wherein, In use, the guide member provides a tapered path to the lumen that guides along at least two perpendicular axes.

6. The retainer according to any one of the preceding claims, wherein, In use, the guide member provides a tapered path that tapers towards the lumen along at least two perpendicular axes.

7. The retainer according to any one of the preceding claims, wherein In use, the guide member is fluidly connected to the end of the fluid conduit.

8. The retainer according to any one of the preceding claims, wherein, The guide member includes a narrow end, and the opening size of the narrow end is substantially 0.5 μm to 50 μm, 1 μm to 25 μm, 2 μm to 20 μm, 3 μm to 15 μm, 3 μm to 10 μm, 4 μm to 6 μm, 4.5 μm to 5.5 μm, or 3 μm to 5 μm.

9. The retainer according to any one of the preceding claims, wherein, The guide member includes a narrow end and a wide end, and: i. The narrow end of the guide member is configured to receive within the lumen of the fluid conduit in use; or ii. The fluid conduit is configured to receive within the narrow end of the guide member in use.

10. The retainer according to any one of the preceding claims, wherein, The guide member includes a sealing element for sealing around the fluid conduit inside or outside the fluid conduit in use.

11. The retainer according to any one of the preceding claims, wherein, The support member has a recess configured to at least partially receive the fluid conduit in use.

12. The retainer according to claim 11, wherein, The lumen of the guide member is substantially concentric with the lumen of the recess.

13. The retainer according to any one of the preceding claims, wherein, The support member includes a holding element for holding the fluid conduit within the holding element in use.

14. The retainer according to any one of the preceding claims, wherein, The support member includes the guide member.

15. The retainer according to any one of the preceding claims, wherein, The support member includes: a first support member located at, near, or including the guide member; and a second support member remote from the guide member.

16. The retainer according to any one of the preceding claims, further comprising the fluid conduit.

17. The retainer according to claim 16, wherein, The fluid conduit includes a needle, syringe, or pipette.

18. The retainer according to claim 17, wherein, The fluid conduit includes a micropipette.

19. The retainer according to any one of claims 16 to 18, wherein The volume of the fluid conduit is substantially 0.5 μl to 50 μl, 2 μl to 30 μl, 5 μl to 25 μl, 10 μl to 20 μl, or 15 μl.

20. The retainer according to any one of claims 16 to 19, wherein, The inner diameter of the fluid conduit is substantially 0.5 μm to 20 μm, 1 μm to 10 μm, 2 μm to 8 μm, 3 μm to 7 μm, 4 μm to 6 μm, 3.5 μm to 5.5 μm, or 4.5 μm to 5.5 μm.

21. The retainer according to any one of claims 16 to 20, wherein When the retainer receives the fluid conduit, the lumen of the guide member is substantially concentric with the lumen of the fluid conduit.

22. A package comprising a fluid conduit holder according to any one of the preceding claims, optionally, the package further comprising a removable protective membrane cover and / or an interchangeable guide.

23. A set of instructions configured to 3D print a fluid conduit holder or a package according to any one of the preceding claims.

24. A method of filling a fluid conduit using a fluid conduit holder, the holder comprising a support and a guide, the method comprising: i. positioning the fluid conduit relative to the guide such that the guide provides a tapered path to the lumen of the fluid conduit; and ii. at least partially filling the fluid conduit with fluid via the guide.

25. A method of in vitro fertilization of an oocyte, comprising the method according to claim 24, the method of in vitro fertilization of an oocyte comprising: i. at least partially filling the fluid conduit with sperm and optionally a working fluid; and further: ii. injecting the sperm into the oocyte by ICSI or piezo-ICSI.

Images