Fluid conduit holder and related methods

Abstract

The fluid conduit holder 1 comprises a support 10 and a guide 20. The support 10 is configured to receive and position the fluid conduit 100 relative to the guide 20 in use, the guide 20 providing a tapered path for the lumen 110 of the fluid conduit 100.

Description

[Technical Field]

[0001] The present invention relates to a fluid conduit holder intended to accommodate a fluid conduit, such as a micropipette, which may be used, for example, for piezo-mediated microinjection or piezo-mediated intracytoplasmic sperm injection (piezo ICSI), to assist in delivering fluids, such as operating fluids (as disclosed in U.S. Patent No. 5,929,499), and / or sperm to the fluid conduit.

[0002] The invention further relates to a method of filling a fluid conduit using a fluid conduit holder, for example for in vitro oocyte fertilization by piezo ICSI. [Background technology]

[0003] Male-attributable infertility accounts for approximately 50% of couples' inability to conceive. In the 1980s, many procedures were developed to address fertilization problems resulting from male gamete dysfunction, including the injection of a single sperm cell (sperm) under the zona pellucida of the oocyte, but the success rate was very limited. However, during subzonal injection of the oocyte, the egg membrane was accidentally breached, allowing the sperm to be delivered into the oocyte cytoplasm. This led to the development of intracytoplasmic sperm injection, a technique still used in humans today.

[0004] Intracytoplasmic sperm injection (ICSI) is an in vitro fertilization procedure in which sperm cells are directly injected into the cytoplasm of an oocyte. ICSI is performed in a culture dish under a microscope by a human operator using a micromanipulator and a micropipette to manipulate the oocyte and sperm. A thin, glass micropipette (holding pipette) stabilizes the mature oocyte with gentle suction applied by a microinjector, while a thin, sharp glass micropipette (injecting micropipette) is used to retrieve a single spermatozoon, immobilized by injuring its tail with the tip of the micropipette. Mechanical force applied by the operator via the micromanipulator causes the injecting micropipette to puncture the oocyte's zona pellucida and membrane. The sperm are then injected into the oocyte's cytoplasm. ICSI has been involved in the birth of more than 2 million babies worldwide. However, ICSI is a labor-intensive technique that requires highly skilled operators, and there is still room for improvement in terms of standardization and success rate of the technique.

[0005] An alternative oocyte microinjection method, called "piezo-mediated" ICSI (piezo ICSI), was developed for mouse oocytes in 1995 because conventional ICSI failed to resolve fertilization issues in mouse IVF (IVF) procedures. Piezo ICSI is similar to conventional ICSI (c-ICSI) in that a micropipette containing a single sperm punctures the oocyte, delivering the sperm into the oocyte cytoplasm. However, whereas in c-ICSI the zona pellucida and chorionic membrane are punctured by mechanical force applied by the operator to the micropipette via a micromanipulator, piezo ICSI uses the piezoelectric effect to puncture the oocyte. The piezoelectric effect is a phenomenon in which certain materials accumulate charge in response to mechanical stress and, conversely, mechanical stress can be induced by the application of electricity. The piezoelectric effect has many technological applications. In piezo ICSI, a short "piezo pulse" is applied to the sperm-containing micropipette, resulting in an ultrafast, submicron forward momentum of the micropipette. This precise and rapid movement is used to puncture the zona pellucida and chorionic membrane in a manner that places less stress on the oocyte than the mechanical forces used in c-ICSI.

[0006] However, ICSI and piezo ICSI remain labor-intensive techniques requiring highly skilled operators, which the present invention seeks to improve. [Prior art documents] [Patent documents]

[0007] [Patent Document 1] International Publication No. 2021094588 Summary of the Invention

[0008] The present invention provides a fluid conduit holder comprising a support and a guide, wherein the support is configured to receive and position a fluid conduit relative to the guide in use, and the guide provides a tapered pathway to the lumen of the fluid conduit.

[0009] The present invention further provides a method of filling a fluid conduit using a fluid conduit holder including 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; ii. at least partially filling the fluid conduit with fluid through the guide; Includes.

[0010] The invention further provides further embodiments as set out in the dependent claims.

[0011] The claimed invention generally provides a subsystem suitable for use in or configured for use in a fluid delivery system, which may form part of a fluid delivery system in general, and in particular a fluid delivery system for in vitro fertilization.

[0012] The claimed invention advantageously assists an operator in delivering fluid (which may contain sperm and / or a working solution for a particular application) into a fluid conduit, such as a micropipette (or microcapillary pipette), which may be made of glass and may be used, for example, for piezo-mediated microinjection or piezo-mediated intracytoplasmic sperm injection (piezo ICSI), which may require significant control and precision. The working solution is typically filled from the rear (rear-filling). Sperm or other cells are usually in a holding medium, and the sperm holding medium is typically inserted from the front. The claimed invention may assist in filling the fluid conduit from one or both ends. The claimed invention may therefore shorten preparation time and reduce the possibility of risks and accidents, thereby reducing waste and improving success rates.

[0013] Fluid conduits can be shipped in holders, reducing the risk of damage to the fragile glass conduits and / or allowing them to be easily removed for attachment to a micromanipulator. [Brief explanation of the drawings]

[0014] Embodiments of the present disclosure will now be described, by way of example only, with reference to the accompanying drawings, in which:

[0015] [Figure 1] FIG. 1 is a schematic perspective view of a first fluid conduit holder according to some embodiments of the present disclosure. [Figure 2] FIG. 2 is a schematic perspective view of a package for a fluid conduit holder according to some embodiments of the present disclosure. [Figure 3a-3b] 3a and 3b are schematic cross-sectional views of an interface between a fluid conduit holder and a fluid conduit according to some embodiments of the present disclosure. [Figure 4] FIG. 4 is a schematic perspective view of a second fluid conduit holder according to some embodiments of the present disclosure. [Figure 5a-5b] 5a and 5b are photographs of a second fluid conduit holder according to some embodiments of the present disclosure. [Figure 6a-6b] 6a and 6b are photographs of a third fluid conduit holder according to some embodiments of the present disclosure. DETAILED DESCRIPTION OF THE INVENTION

[0016] Aspects and features of particular examples and embodiments are described herein. Some aspects and features of particular examples and embodiments can be implemented conventionally, and are not described in detail for the sake of brevity. Thus, it will be understood that aspects and features of the devices and methods described herein that are not described in detail can be implemented in accordance with any suitable conventional techniques.

[0017] FIG. 1 is a schematic perspective view of a first fluid conduit holder 1 according to some embodiments of the present disclosure. FIG. 1 illustrates the fluid conduit holder 1. The fluid conduit holder 1 includes three axially aligned rectangular block supports 10a, 10b, and 10c for receiving and supporting a fluid conduit 100, and a guide 20. The fluid conduit 100 may include a needle, a syringe, or a pipette (particularly a micropipette). However, the fluid conduit 100 itself is not the key to the present invention. The multiple supports 10 include a first support 10a distal to the guide 20, and second and third supports 10b and 10c. Here, 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 the example of Figure 1, the supports 10 each include a channel-like axial recess (not shown) for receiving a portion of the fluid conduit 100 and are secured to the base 30 of a protective seal package for the holder 1, which will be described below with reference to Figure 2.

[0018] 1 includes a funnel 20 adjacent to the third support 10c. When the holder 1 receives a fluid conduit 100 in or on the support 10 during use, the support 10 positions the fluid conduit 100 relative to the guide 20, which provides a tapered (narrowed) path to the lumen 110 of the fluid conduit 100. The guide 20 is configured to guide / direct a user inserting a container containing a fluid to be backfilled into the lumen 110 of the fluid conduit 100 so that the container can be easily inserted into the lumen 110 and the user can fill the fluid conduit 100 with fluid from the container.

[0019] Guide 20 includes a tapered guide that provides a narrowing path to lumen 110 of fluid conduit 100. Preferably, guide 20 is configured to provide a tapered path that guides toward lumen 110 in at least two perpendicular axes. In FIG. 1 , funnel guide 20 tapers toward lumen 110 in two perpendicular axes (i.e., the y-axis and z-axis, which are perpendicular to the x-axis along which fluid conduit 100 extends), thereby providing a path to lumen 110 that is guided toward (or constrained away from) lumen 110 in four Cartesian degrees of freedom (i.e., ±y and ±z). In other words, an object in funnel 20, such as a container inserted into the lumen of fluid conduit 100, is constrained in four Cartesian degrees of freedom to move within funnel 20 and is thus guided toward lumen 110.

[0020] 1 is also configured to be axially aligned with the fluid conduit 100 when the fluid conduit 100 is received in the holder 1, with the lumen of the funnel guide 20 being substantially concentric with the lumen 110 of the fluid conduit 100. In embodiments where the support 10 includes a recess for receiving the fluid conduit 100, the lumen of the guide 20 can be substantially concentric with the lumen of the recess to provide suitable alignment when the fluid conduit 100 is received therein.

[0021] 1 , the guide comprises a funnel 20 that tapers in two perpendicular axes and advantageously guides (restricts) movement in four Cartesian (x, y, z) degrees of freedom (allowing only free axial movement through the funnel 20 and restricting movement away from the lumen 110 when in proximity to the funnel 20). More generally, the guide 20 may comprise a funnel or cone, or a conical, frustoconical, concave, convex, or tapered surface, such as a portion of a funnel or cone (e.g., a half-funnel). In some embodiments, the guide 20 provides a tapered path that guides toward the lumen 110 in at least two perpendicular axes and / or provides a path to the lumen 110 that is guided toward (or restricted away from) the lumen 110 in at least three Cartesian degrees of freedom.

[0022] The guide 20 may be a separate element from the support 10, as shown in FIG. 1, or may be integrated into one or more of the supports 10, as shown in FIGS. 6a and 6b. The guide 20 is appropriately sized to fit the fluid conduit 100 to be used; this application is particularly relevant to holders suitable for micro-sized fluid conduits 100, such as P10, P20, and P50 micropipettes, which are suitable for volumes of 0.5-10 μl, 2-20 μl, and 5-50 μl, respectively. For piezo ICSI, the volume of working fluid backfilled into the micropipette is usually 10-20 μl. In some embodiments, the fluid conduit 100 may have a volume of substantially 0.5-50 μl, 2-30 μl, 5-25 μl, 10-20 μl, or 15 μl, and / or an inner diameter of substantially 0.5-20 μm, 1-10 μm, 2-8 μm, 3-7 μm, 4-6 μm, 3.5-5.5 μm, or 4.5-5.5 μm. In such embodiments, the guide 20 may include a narrow end having opening dimensions (e.g., diameter in the case of a funnel, width or height in the case of a curved surface) that are substantially the same as the inner diameter of the fluid conduit 100 in use, or may be slightly larger (to aid insertion but not unduly restrict the inner diameter of the fluid conduit 100) or slightly smaller (potentially providing a direct pathway into the lumen 110 of the fluid conduit 100). In some embodiments, the opening size at the narrow end of the guide 20 may therefore be substantially between 0.5 and 50 μm, 1 and 25 μm, 2 and 20 μm, 3 and 15 μm, 3 and 10 μm, 4 and 6 μm, 4.5 and 5.5, or 3 and 5 μm.

[0023] During an ICSI procedure, sperm-holding medium is aspirated from the front end, typically about 2–10 μl. In piezo ICSI, the micropipette is first backfilled with working fluid, typically about 15 μl, and then drawn down to the tip of the front end. After this, sperm-holding medium is aspirated from the tip of the front end, bringing it into direct contact with the working fluid.

[0024] That is, the present invention assists an operator in filling a fluid conduit 100 by providing a fluid holder 1 that, when in use, presents a tapered pathway to the lumen 110 of the fluid conduit 100 with the fluid conduit positioned in the holder 1. In use, the operator positions the fluid conduit 100 relative to the guide 20 (e.g., substantially abutting 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 presents a tapered pathway to the lumen 110 of the fluid conduit 100, assisting in the insertion of a container containing a fluid to be filled into the lumen 110 of the fluid conduit 100. In some embodiments, the guide 20 may be fluidly connected to one end of the fluid conduit 100 when in use.

[0025] 2 is a schematic diagram of packaging for a fluid conduit holder 1 according to some embodiments of the present disclosure. The package includes a base 30, a sidewall 35, and a removable protective peel-off film lid 40. The removable protective peel-off film lid 40 covers the support 10 and guide 20 therein. The package can provide a safe and sterile housing for the holder 1. An operator removes the film lid 40 to reveal the holder 1 ready for use.

[0026] 3a and 3b are schematic cross-sectional views of an interface between a fluid conduit holder 1 and a fluid conduit 100 according to some embodiments of the present disclosure. As shown in FIGS. 3a and 3b, in use, in some embodiments, the fluid conduit 100 may be received in the guide 20 as in FIG. 3a, or the guide 20 may be received in the fluid conduit 100 as in FIG. 3b. The guide 20 is sized appropriately for the intended fluid conduit 100 and intended interfacing method. The guide 20 may be replaceable / substitute, and multiple guides 20 of different sizes may be provided, for example, in a package. In some embodiments, the fluid conduit 100 and / or the guide 20 are keyed to suitable (interior / exterior) surfaces to provide a secure engagement mechanism. In FIG. 3a, the fluid conduit 100 is received in the narrow end of the guide 20, which abuts the exterior of the fluid conduit 100 or, optionally, is secured, for example, by a compressive interference fit. As shown in Figure 3a, guide 20 (alternatively, fluid conduit 100) may include a sealing element 60, such as an O-ring, configured to assist in a fluid seal between fluid conduit 100 and guide 20. In Figure 3b, guide 20 is received within lumen 110 of fluid conduit 100. As shown in Figure 3b, guide 20 may include a sealing element 60, such as a compressible and impermeable material, on an outer surface thereof to assist in a fluid seal between fluid conduit 100 and guide 20. Alternatively, fluid conduit 100 may be provided with a sealing element 60 on its inner wall.

[0027] FIG. 4 is a schematic perspective view of a second fluid conduit holder 1 according to some embodiments of the present disclosure. The main differences from the first embodiment of FIG. 1 are outlined here. In this second embodiment, the holder 1 includes only two supports 10: a first support 10a distal to the guide 20 for supporting the distal end of the fluid conduit 100, and a second support 10b for supporting both the proximal end of the fluid conduit 100 and the guide 20 during use. In this embodiment, the first support 10a extends axially beyond the length of the fluid conduit 100 and provides a recess. The recess includes a retaining member 70 (see inset cross-sectional view) for retaining the fluid conduit 100 therein. The retaining member 70 may include a flexible and / or resilient material. This material limits movement of the fluid conduit 100 in the recess and holds it in place. Having the first support portion 10a extend beyond the axial length of the end of the fluid conduit 100 helps to protect the distal end of the fluid conduit 100 during use, which may be sharp.

[0028] In the embodiment of Figure 4, guide 20 includes a concave surface 20 whose width along the y-axis tapers (narrows) towards lumen 110. The inset cross-sectional view shows that guide 20 is concave in a cross-section perpendicular to the elongation axis of fluid conduit 100, effectively functioning as a half-funnel. Figures 5a and 5b are photographs of the second fluid conduit holder 1, detailing concave guide 20. These images show that guide 20 is configured to guide in two perpendicular axes that, in use, provide a pathway to lumen 110, a pathway that is guided (constrained) in three Cartesian degrees of freedom. First, as shown best in the inset of FIG. 4 and in FIG. 5a, the guide 20 is guided by a tapered, concave shape that narrows in width (along the y-axis) as it approaches the lumen 110 (along either the x- or z-axis), thus guiding (restricting) movement in both y-degrees of freedom; and second, by the axial extension of the guide 20 along the x-axis. The guide 20 may be horizontal, as in FIG. 4, or angled toward the fluid conduit 100 in the z-axis (as shown in FIG. 5a), either way guiding (restricting) movement in the negative z-degree of freedom only. The guide 20 of FIGS. 5a and 5b therefore effectively acts like a half-funnel, restricting movement only in the negative z-degree of freedom, without restricting movement in the positive z-degree of freedom.

[0029] 6a and 6b are photographs of a third fluid conduit holder according to some embodiments of the present disclosure. In this embodiment, the holder 1 includes a first support 10a that is integrated into the package base 30 and distal to a second support 10b. The second support 10b includes a guide 20, both of which have a V-shaped channel recess. As best shown in FIG. 6b, the guide 20 includes a conical recess at the proximal axial end of the support 10, effectively providing an integrated funnel. In use, when a fluid conduit 100 is placed into the channel recess of the support 10, the guide 20 tapers in two perpendicular axes, thereby providing a path for the fluid conduit 100 to enter the lumen 110, guiding it toward the lumen 110 in two perpendicular axes. Thus, similar to FIG. 1, the conical recess tapers in two perpendicular axes, providing a path for the lumen 110 that is guided (bounded) in four Cartesian degrees of freedom.

[0030] The holder 1 also includes an upright portion 50 in the form of a wall that, like the first support portion 10a, is integral with the package base 30. The upright wall 50 positions and holds the second support portion 10b and may additionally support the protective tear-off lid 40 of the package for the holder 1 shown in FIG.

[0031] In some embodiments, guide 20 is configured to be axially aligned with fluid conduit 100 when conduit 100 is received in holder 1. In other embodiments, guide 20 further comprises a flexible tube, such as a telescoping tube or bellows (not shown), that allows guide 20 to move (and / or extend) out of axial alignment with fluid conduit 100 to adjust the angle of entry into guide 20, further assisting the user.

[0032] The various embodiments described herein are presented solely to aid in understanding and teaching the claimed features. These embodiments are provided only as a representative sample of embodiments and are not intended to be exhaustive and / or exclusive. It is understood that the advantages, embodiments, examples, functions, features, structures, and / or other aspects described herein should not be construed as limitations on the scope of the invention as defined by the claims or on the equivalents of the claims, and that other embodiments may be utilized and modifications may be made without departing from the scope of the claimed invention.

[0033] Various embodiments of the present invention may suitably comprise, consist of, or consist essentially of any suitable combination of the disclosed elements, components, features, parts, steps, means, etc., other than those specifically described herein. In addition, the present disclosure may include other inventions not currently claimed but which may be claimed in the future. Protection may also be sought for any features disclosed in one or more of the publications referenced herein in combination with the present disclosure. [Explanation of symbols]

[0034] Any reference signs in the claims shall have no effect on the scope of the claims but are merely for the aid of the reader.

[0035] 1 Fluid conduit holder 10 Support part 20 Guide 30 Package base 35 Package sidewall 40 packaging film 50 Upright part 60 sealing elements 70 Retention elements 100 Fluid conduit 110 Fluid conduit lumen

Claims

1. 1. A fluid conduit holder including a support and a guide, A fluid conduit holder, wherein the support is configured to receive and position a fluid conduit relative to the guide in use, the guide providing a tapered path to the lumen of the fluid conduit.

2. The holder of claim 1 , wherein the guide comprises a funnel or cone, or comprises a conical, frustoconical, concave, convex or tapered surface.

3. 3. A holder according to claim 1 or 2, wherein in use the guide provides a tapered path that guides into the lumen in at least three Cartesian degrees of freedom.

4. 4. A holder according to any one of claims 1 to 3, wherein in use the guide provides a tapered path that guides into the lumen in four Cartesian degrees of freedom.

5. 5. A holder according to any one of claims 1 to 4, wherein in use the guide provides a tapered path leading to the lumen in at least two perpendicular axes.

6. 6. A holder according to any one of claims 1 to 5, wherein in use the guide presents a tapered path which tapers towards the lumen in at least two perpendicular axes.

7. A holder according to any preceding claim, wherein, in use, the guide is fluidly connected to one end of the fluid conduit.

8. 8. The holder of any one of claims 1 to 7, wherein the guide comprises a narrow end having an opening dimension of substantially 0.5 to 50 μm, 1 to 25 μm, 2 to 20 μm, 3 to 15 μm, 3 to 10 μm, 4 to 6 μm, 4.5 to 5.5 or 3 to 5 μm.

9. the guide includes a narrow end and a wide end; i. the narrow end of the guide is configured to be received within the lumen of the fluid conduit in use; or ii. A holder according to any one of claims 1 to 8, wherein the fluid conduit is configured to be received within the narrow end of the guide in use.

10. 10. A holder according to any one of claims 1 to 9, wherein the guide includes a sealing element that seals the interior or exterior of the fluid conduit in use.

11. 11. A holder according to any preceding claim, wherein the support portion has a recess therein configured to at least partially receive the fluid conduit in use.

12. The holder of claim 11 , wherein the guide bore is substantially concentric with the recess bore.

13. A holder according to any preceding claim, wherein the support comprises a retaining member that retains the fluid conduit therein in use.

14. The holder of claim 1 , wherein the support comprises the guide.

15. The support portion is a first support at, near, or including the guide; a second support distal to the guide; and 15. The holder of any one of claims 1 to 14, comprising:

16. The holder of claim 1 , further comprising the fluid conduit.

17. The holder of claim 16 , wherein the fluid conduit comprises a needle, a syringe, or a pipette.

18. 20. The holder of claim 17, wherein the fluid conduit comprises a micropipette.

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

20. 20. The holder of any one of claims 16 to 19, wherein the fluid conduit has an internal diameter of substantially 0.5 to 20 μm, 1 to 10 μm, 2 to 8 μm, 3 to 7 μm, 4 to 6 μm, 3.5 to 5.5 μm, or 4.5 to 5.5 μm.

21. 21. The holder of any one of claims 16 to 20, wherein the lumen of the guide is substantially concentric with the lumen of the fluid conduit when the holder receives the fluid conduit.

22. 22. A package comprising the fluid conduit holder of any one of claims 1 to 21, optionally further comprising a removable protective film lid and / or a replaceable guide.

23. 23. A set of instructions configured to 3D print the fluid conduit holder or package of any one of claims 1 to 22.

24. 1. A method of filling a fluid conduit using a fluid conduit holder, the holder including 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; ii. at least partially filling the fluid conduit with fluid through the guide; A method comprising:

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

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