Hollow-lid tubes for diverse applications

Abstract

A hollow-lid tube is provided with a body extending along a longitudinal axis and a lid configured to fit thereon. A seal is positioned within the tube to align a cavity within the body with one or more holes in the lid, and prevent a sample from escaping the cavity other than by those holes. Additionally, a membrane is positioned to separate the volume of the cavity and an environment surrounding the hollow-lid tube, for example by positioning the membrane within the hole itself, between the body and the lid, on the lid over the hole, etc. Samples to be tested or treated can be provided to a plurality of these tubes and subsequently positioned within a testing solution such that the sample and the solution each contact the membrane. Thus, simultaneous testing and / or assaying of one or more samples can be performed, greatly reducing analysis time and material costs.

Description

HOLLOW-LID TUBES FOR DIVERSE APPLICATIONSCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the priority benefit of U.S. Provisional Patent Applications No. 63 / 441,262, filed January 26, 2023, 63 / 448,045, filed February 24, 2023, and 63 / 625,344, filed January 26, 2024, the entireties of which are explicitly incorporated by reference herein.STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH AND DEVELOPMENT

[0002] This invention was made with U.S. Government support under award numbers AG20618 and AR071681, awarded by the National Institutes of Health. The government has certain rights in the invention.BACKGROUND

[0003] Laboratory equipment is integral to the performance of life sciences research, including biochemistry, molecular biology, genetics, pharmacology, etc. Microcentrifuge tubes are widely used in industry to facilitate protocols for myriad applications, including PCR reactions, centrifugation, sample preparation, analysis, reagent storage, etc. However, the role of these tubes in these protocols is typically restricted to that of a temporary storage vessel until more specialized equipment is acquired and implemented, e.g., short-term storage in the case of analysis as samples are provided to a separate analysis medium, long-term storage in the case of sample preparation where the sample must be held until such time the sample and the rest of the experiment is ready, etc. Having served such basic functions, the tubes are typically discarded.

[0004] What is desired, therefore, are tubes that provide improved functionality to increase the efficiency of protocols and / or throughout of samples particularly in laboratory settings, such as during dialysis, the development and modification of biomaterials including cells and tissues, etc.SUMMARY

[0005] Aspects of the present disclosure are directed to a hollow-lid tube. In some embodiments, the hollow-lid tube includes a body extending along a longitudinal axis. In someembodiments, the body has a closed end, an open end, and a cavity extending from the closed end to the open end. In some embodiments, the hollow-lid tube includes a lid. In some embodiments, the lid includes at least one hole therethrough and in fluid communication with the cavity and a seal. In some embodiments, the seal is configured to maintain the body and the hole in a desired alignment along the longitudinal axis. In some embodiments, the hollow-lid tube includes a membrane separating the cavity from an exterior surface of the tube.

[0006] In some embodiments, a first hole is concentric with the cavity when the lid is positioned on the body. In some embodiments, the seal surrounds the first hole. In some embodiments, the membrane is positioned within the hole. In some embodiments, the membrane is a dialysis membrane. In some embodiments, the membrane is integrated with the seal. In some embodiments, the lid is hingedly attached to the body. In some embodiments, the open end and the hole have substantially the same diameter. In some embodiments, the open end has an inner diameter surface and the seal includes a friction fit between the lid and the inner diameter surface.

[0007] Aspects of the present disclosure are directed to a hollow-lid tube including a body extending along a longitudinal axis, wherein the body has an inner diameter surface, a closed end, an open end, and a cavity extending from the closed end to the open end; and a lid including a seal configured to maintain alignment of the lid and the body via a friction fit between the lid and the inner diameter surface; at least one hole positioned within the seal, the hole providing fluid communication between the cavity and an environment exterior to the cavity; and a dialysis membrane positioned within the hole. In some embodiments, the lid is hingedly attached to the body.

[0008] Aspects of the present disclosure are directed to a method of performing dialysis. In some embodiments, the method includes providing one or more hollow-lid tubes. As discussed above, in some embodiments, the hollow-lid tubes include a body extending along a longitudinal axis, wherein the body has a closed end, an open end, and a cavity extending from the closed end to the open end; a lid including at least one hole therethrough and in fluid communication with the cavity; and a seal, wherein the seal is configured to maintain the body and the hole in a desired alignment along the longitudinal axis, and a membrane.

[0009] In some embodiments, the membrane is positioned within the hole. In some embodiments, the membrane is integrated with the seal. In some embodiments, the lid is hingedly attached to the body. In some embodiments, the open end and the hole have substantially the same diameter. In some embodiments, the open end has an inner diametersurface and the seal includes a friction fit between the lid and the inner diameter surface. In some embodiments, the one or more hollow-lid tubes includes a multiwell microtiter plate as the body, a substrate including an array of holes corresponding to wells in the multiwell microtiter plate, and one or more membranes positioned between the holes and wells in the multiwell microtiter plate.

[0010] In some embodiments, the method includes providing a sample to the cavity, attaching the lid to the body such that the membrane separates the cavity from an exterior surface of the tube, and positioning the tube in a volume of a solution such that the sample and the solution are both in contact with the membrane. In some embodiments, the sample includes a concentration of bone powder in a bone protein extraction solution.BRIEF DESCRIPTION OF THE DRAWINGS

[0011] The drawings show embodiments of the disclosed subject matter for the purpose of illustrating the invention. However, it should be understood that the present application is not limited to the precise arrangements and instrumentalities shown in the drawings, wherein:

[0012] FIGs. 1A-1F are schematic representations of hollow-lid tubes according to some embodiments of the present disclosure;

[0013] FIG. 2 is a chart of a method of performing a protocol such as dialysis utilizing hollow-lid tubes according to some embodiments of the present disclosure; and

[0014] FIGs. 3A-3C is a schematic representation of performing a protocol such as dialysis, keeping cells, separation of cell organelles, small-scale preliminary tests, etc., utilizing hollow-lid tubes according to some embodiments of the present disclosure.DETAILED DESCRIPTION

[0015] Referring now to FIGs. 1 A-1F, some embodiments of the present disclosure are directed to a hollow-lid tube 100. In some embodiments, hollow-lid tube 100 includes at least one body 102 and at least one lid 104. In some embodiments, body 102 extends along at least a longitudinal axis L. The embodiments of the present disclosure will be described with bodies 102 having a generally cylindrical shape, as would be particularly suited for use in microcentrifuge applications. However, the present disclosure is not intended to be limited in this regard, as body 102 may be any suitable cross-sectional shape, e.g., triangular, rectangular, etc. Further, the embodiments of the present disclosure will be described with longitudinalaxes L that are larger than a corresponding latitudinal axis (not pictured), however the present disclosure is not intended to be limited in this regard, as body 102 may also extend spatially along axes other than longitudinal axis L, e.g., a latitudinal axis.

[0016] Still referring to FIGs. 1 A-1F, in some embodiments, body 102 includes a closed end 102A and an open end 102B. In some embodiments, body 102 includes a cavity 102C. In some embodiments, cavity 102C extends from closed end 102 A. In some embodiments, cavity 102C extends from closed end 102 A to open end 102B. Cavity 102C has any suitable volume to hold a sample S of a desired size and / or volume, as will be discussed in greater detail below. In some embodiments, open end 102B and / or body 102 have an inner diameter surface 102S.

[0017] In some embodiments, lid 104 is configured to inhibit sample S from leaving cavity 102C. In some embodiments, lid 104 is configured to attach to body 102 at or adjacent to open end 102B. Lid 104 attaches to body 102 via any suitable mechanism, e.g., a threaded fit, a friction fit, etc., or combinations thereof, as will be discussed in greater detail below. In some embodiments, lid 104 reversibly attaches to body 102. In some embodiments, lid 104 is hingedly attached to body 102.

[0018] In some embodiments, lid 104 includes at least one hole 104H. In some embodiments, hole 104H extends through lid 104. The embodiments of the present disclosure will be described with holes 104H having a generally circular shape. However, the present disclosure is not intended to be limited in this regard, as holes 104H may be any suitable cross- sectional shape, e.g., triangular, rectangular, etc. In some embodiments, lid 104 includes a plurality of holes 104H (not pictured).

[0019] In some embodiments, tube 100 includes one or more seals 104S. In some embodiments, seal 104S is configured to maintain body 102 and hole 104H in a desired alignment. In some embodiments, this desired alignment is along longitudinal axis L. In some embodiments, seal 104S is configured to inhibit sample S from leaving cavity 102C. In some embodiments, lid 104 includes one or more seals 104S. In some embodiments, seal 104S surrounds hole 104H. In some embodiments, seal 104S is positioned on body 102, rather than lid 104. In some embodiments, seal 104S includes structural components positioned on body 102 and lid 104 that, in combination, provide the desired alignment between body 102 and hole 104H, inhibition to movement of sample S from cavity 102C, etc., or combinations thereof. In some embodiments, seal 104S includes one or more gaskets. In some embodiments,seal 104S is a friction fit between lid 104 and inner diameter surface 102S, as will be discussed in greater detail below.

[0020] Referring now specifically to FIGs. 1C-1D, in some embodiments, holes 104H are in fluid communication with cavity 102C at least when lid 104 is positioned on body 102. In some embodiments, holes 104H are in fluid communication with cavity 102C and an environment surrounding hollow-lid tube 100, e.g., environment E. In some embodiments, hole 104H and cavity 102C are concentric or substantially concentric when lid 104 is positioned on body 102. In some embodiments, open end 102B and hole 104H have substantially the same diameter. In some exemplary embodiments, lid 104 includes an annular protrusion 104P extending therefrom, with hole 104H positioned within the protrusion. Annular protrusion 104P is sized so that it may be inserted into cavity 102C, forming a friction fit with inner diameter surface 102S, reversibly maintaining alignment between hole 104H and cavity 102C, and functioning as seal 104S to prevent sample S from escaping the cavity except via the hole.

[0021] In some embodiments, hollow-lid tube 100 includes one or more membranes 106. In some embodiments, hollow-lid tube 100 includes a plurality of membranes 106. In some embodiments, membrane 106 is positioned to separate cavity 102C from environment E, e.g., an exterior surface of tube 100, when lid 104 is positioned on body 102. In some embodiments, membrane 106 is positioned on lid 104. In some embodiments, membrane 106 is positioned within hole 104H. In some embodiments, membrane 106 is positioned on body 102. In some embodiments, membrane 106 is positioned at open end 102B. In some embodiments, membrane 106 is positioned within cavity 102C. In some embodiments, membrane 106 is integrated with seal 104S. In some embodiments, membrane 106 is any suitable composition for performing the desired separation therewith. In some embodiments, membrane 106 is commercially available, made by investigator / researcher, or combinations thereof. In some embodiments, membrane 106 includes a molecular sieve, e.g., as applied for molecular filtration (cell organelles). In some embodiments, the molecular sieve includes a net with holes within nanometer holes, micrometer holes, or combinations thereof. In some embodiments, membrane 106 is a dialysis membrane.

[0022] In some embodiments, body 102 and / or lid 104 are composed of any suitable material or combination of materials to hold a desired volume of sample S. In some embodiments, body 102 and / or lid 104 are composed of one or more polymers, glasses, or combinations thereof, e.g., polypropylene, polyethylene, borosilicate glass, etc. Exemplary embodiments of hollow-lid tubes 100 are provided in Table 1 below:Table 1 : Lid and Body Sizes of Exemplary Hollow-Lid Tubes

[0023] Referring specifically to FIGs. IE- IF, in some embodiments, tube 100 includes a clasping mechanism 108. In some embodiments, clasping mechanism 108 is configured to reversibly grip body 102 to inhibit lid 104 from disconnecting from the body, e.g., from open end 102B during handling of tube 100. In some embodiments, clasping mechanism 108 includes one or more hooks.

[0024] Some embodiments of the present disclosure are directed to a procedure for making a hollow-lid tube, e.g., tube 100. In some embodiments, a rod or tube having the desired diameter of the hole is heated. In some exemplary embodiments, a glass tube, e.g., a Pasteur pipette, is heated in a flame of Bunsen burner. The heated rod / tube is then contacted with the lid of an Eppendorf-type tube of the selected size, melting the hole in the center thereof. In some embodiments, the lid edge is smooth. In some embodiments, the hollow-lid tubes can be made during production of the microcentrifuge tubes. In some embodiments, a membrane is positioned between the cavity of the body and the hole in the lid, e.g., a membrane is positioned over the open end of the tube, and the lid is closed to hold the membrane in place.

[0025] Referring now to FIG. 2, some embodiments of the present disclosure are directed to a method 200 of performing a protocol, e.g., dialysis. At 202, one or more hollow-lid tubes are provided. As discussed above, in some embodiments, the hollow-lid tubes include at least a body, a lid, and a membrane. In some embodiments, the body of the hollow-lid tubes extends along a longitudinal axis. In some embodiments, the body has a closed end, an open end, and a cavity extending from the closed end to the open end. In some embodiments, the lid includes at least one hole therethrough and in fluid communication with the cavity. In some embodiments, the hollow-lid tubes include, e.g., on the lid, body, or combination thereof, a seal. In someembodiments, the seal is configured to maintain the body and the hole in a desired alignment along the longitudinal axis, configured to inhibit a sample from leaving the cavity, or combinations thereof. In some embodiments, the membrane is positioned within the hole. In some embodiments, the membrane is integrated with the seal.

[0026] Still referring to FIG. 2, at 204, a sample is provided to the cavity. In some embodiments, the sample includes a concentration of bone powder in a bone protein extraction solution. At 206, the lid is attached to the body such that the membrane separates the cavity from an exterior surface of the tube.

[0027] In some embodiments, the selected experiment can then be conducted. At 208, the tube is positioned in a volume of a solution such that the sample and the solution are both in contact with the membrane. In an exemplary embodiment, after filling the tube with the sample, the mouth of the tube is covered with a membrane, e.g., any type undergoing the investigation or a dialysis membrane, the hollow-lid tubes are closed (allowing the membrane to line the hole), the tubes are placed, e.g., into a rack, and the rack with the tubes placed into a beaker with a buffer. For high-throughput analyses, microtiter-plates of various sizes can be used to make holes at the bottom of each well (to close the plate, a plate similar to a holder for pipette tips can be used). In an exemplary embodiment, the hollow-lid tubes includes a multiwell microtiter plate as the body, a substrate including an array of holes corresponding to wells in the multiwell microtiter plate, and one or more membranes positioned between the wells in the multiwell microtiter plate and an environment surrounding the plate, e.g., via positioning in the array of holes, positioning between the substrate and the wells, positioning such that the substrate is between the membrane and the plate, etc., or combinations thereof.

[0028] Referring now to FIGs. 3 A-3C, hollow-lid tubes consistent with embodiments of the present disclosure enable high-throughput testing and analysis of target samples. For example, referring specifically to FIG. 3 A, a plurality of hollow-lid tubes 100 can be provided, each with a dialysis membrane separating the sample held within the tubes from a surrounding solution, e.g., buffer B. Simultaneous testing and / or assaying of one or more samples S can greatly reduce analysis time, as well as material costs as membrane surface area per sample is reduced. In addition to dialysis processes, e.g., see FIG. 3A, the hollow-lid tubes are further particularly beneficial for the development and testing of membranes themselves, e.g., those prepared for in vivo implantation after in vitro tests; the development and testing of biomaterials or implants; modification and analysis of cells and tissues (microscale); extraction and / or dialysis of bone matrix proteins as well as proteins from other tissues of humans, animals, andplants; keeping cells, for example, before single-cell analysis, i.e., single-cell genomics, singlecell proteomics, metabolomics, etc., cells are kept on a membrane and / or separation of cell organelles of any type of eukaryotic organism, phytoplankton organisms, e.g., environmental tests and research, fisheries, agriculture, etc. using molecular sieve with or instead of membranes, e.g., see FIG. 3B; small-scale preliminary and / or high-throughput tests of synthetic and natural membranes for desalination of water and other solutions, e.g., see FIG. 3C, etc.

[0029] Although the invention has been described and illustrated with respect to exemplary embodiments thereof, it should be understood by those skilled in the art that the foregoing and various other changes, omissions and additions may be made therein and thereto, without parting from the spirit and scope of the present invention.

Claims

CLAIMSWhat is claimed is:

1. A hollow-lid tube, comprising: a body extending along a longitudinal axis, wherein the body has a closed end, an open end, and a cavity extending from the closed end to the open end; and a lid including: at least one hole therethrough and in fluid communication with the cavity, and a seal, wherein the seal is configured to maintain the body and the hole in a desired alignment along the longitudinal axis.

2. The hollow-lid tube according to claim 1, wherein a first hole is concentric with the cavity when the lid is positioned on the body.

3. The hollow-lid tube according to claim 2, wherein the seal surrounds the first hole.

4. The hollow-lid tube according to claim 1, further comprising a membrane separating the cavity from an exterior surface of the tube.

5. The hollow-lid tube according to claim 4, wherein the membrane is positioned within the hole.

6. The hollow-lid tube according to claim 4, wherein the membrane is a dialysis membrane.

7. The hollow-lid tube according to claim 4, wherein the membrane is integrated with the seal.

8. The hollow-lid tube according to claim 1, wherein the lid is hingedly attached to the body.

9. The hollow-lid tube according to claim 1, wherein the open end and the hole have substantially the same diameter.

10. The hollow-lid tube according to claim 1, wherein the open end has an inner diameter surface and the seal includes a friction fit between the lid and the inner diameter surface.

11. A method of performing dialysis, comprising: providing one or more hollow-lid tubes, comprising: a body extending along a longitudinal axis, wherein the body has a closed end, an open end, and a cavity extending from the closed end to the open end; a lid including: at least one hole therethrough and in fluid communication with the cavity; and a seal, wherein the seal is configured to maintain the body and the hole in a desired alignment along the longitudinal axis, and a membrane, providing a sample to the cavity; attaching the lid to the body such that the membrane separates the cavity from an exterior surface of the tube; and positioning the tube in a volume of a solution such that the sample and the solution are both in contact with the membrane.

12. The method according to claim 11, wherein the membrane is positioned within the hole.

13. The method according to claim 11, wherein the membrane is integrated with the seal.

14. The method according to claim 11, wherein the lid is hingedly attached to the body.

15. The method according to claim 11, wherein the open end and the hole have substantially the same diameter.

16. The method according to claim 11, wherein the open end has an inner diameter surface and the seal includes a friction fit between the lid and the inner diameter surface.

17. The method according to claim 11, wherein the sample includes a concentration of bone powder in a bone protein extraction solution.

18. The method according to claim 11, wherein the one or more hollow-lid tubes includes a multiwell microtiter plate as the body, a substrate including an array of holes corresponding to wells in the multiwell microtiter plate, and one or more membranes positioned between the holes and wells in the multiwell microtiter plate.

19. A hollow-lid tube, comprising: a body extending along a longitudinal axis, wherein the body has an inner diameter surface, a closed end, an open end, and a cavity extending from the closed end to the open end; and a lid including: a seal configured to maintain alignment of the lid and the body via a friction fit between the lid and the inner diameter surface; at least one hole positioned within the seal, the hole providing fluid communication between the cavity and an environment exterior to the cavity; and a dialysis membrane positioned within the hole.

20. The hollow-lid tube according to claim 19, wherein the lid is hingedly attached to the body.

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