ONE-TOUCH DEVICE FOR COLLECTING FLUID

Abstract

The present invention relates to a device for collecting body fluids including (a) a fluid receiving part including a fluid receiving space in which an internal low pressure less than an external atmospheric pressure is formed; (b) a fluid collection part including a hollow microstructure connected to form an open system with the fluid receiving space of the fluid receiving part; and (c) a microstructure hollow barrier for blocking the hollow of the microstructure to maintain the internal low pressure formed in the fluid receiving means.

Claims

1. A device for collecting body fluids comprising: (a) a fluid receiving part comprising a fluid receiving space in which an internal low pressure less than an external atmospheric pressure is formed; (b) a fluid collection part comprising a hollow microstructure connected to form an open system with the fluid receiving space of the fluid receiving part; and (c) a microstructure hollow barrier for blocking a hollow of the microstructure to maintain the internal low pressure formed in the fluid receiving means.

2. The device according to claim 1, wherein the fluid receiving part comprises (a) a porous structure having air permeability and (b) a coating agent for coating an exterior of the porous structure such that the porous structure has airproof capability.

3. The device according to claim 1, wherein the microstructure hollow barrier is a polymer coating formed by a microstructure dipping method.

4. The device according to claim 3, wherein the polymer coating is composed of a biocompatible and biodegradable polymer.

5. The device according to claim 1, wherein the microstructure hollow barrier is a polymer cap formed by a microstructure capping method.

6. The device according to claim 5, wherein, when body fluids are collected from a subject, the polymer cap is separated from a microstructure-tip to open a hollow and form a pressure gradient when a vertical force is applied to cause the microstructure to penetrate a skin barrier of the subject.

7. The device according to claim 5, wherein the polymer cap forms an exposed-tip capping structure.

8. The device according to claim 5, wherein, when body fluids are collected from a subject, the polymer cap is separated from a microstructure-tip and does not enter a skin barrier of the subject.

9. The device according to claim 1, wherein the fluid receiving part has transparency.

10. An integrated analysis system for body fluids comprising the device for collecting body fluids according to any one of claims 1 to 9 and a device for analyzing collected fluids.

11. The integrated analysis system according to claim 10, wherein the device for analyzing collected fluids comprises a signal device for generating a fluid analysis signal.

Description

DESCRIPTION OF DRAWINGS

[0041] FIG. 1 illustrates one embodiment of a device for collecting body fluids. 1: FLUID RECEIVING PART; 2: FLUID COLLECTION PART; 3: HOLLOW BARRIER; 11: FLUID RECEIVING PART OUTER WALL; 12: FLUID RECEIVING SPACE; 21: HOLLOW MICROSTRUCTURE; 22: SUPPORT.

[0042] FIG. 2 illustrates one embodiment of a device for collecting body fluids including a polymer coating which is a hollow barrier formed by a dipping method. 31: POLYMER COATING.

[0043] FIG. 3 illustrates one embodiment of a device for collecting body fluids including a polymer cap which is a hollow barrier formed by a capping method. 32: POLYMER CAP.

[0044] FIG. 4 illustrates one embodiment of the distal end of a hollow microstructure.

[0045] FIG. 5 is a schematic diagram showing one embodiment of a device for collecting body fluids. FIG. 5a is an exploded view showing each part of a device for collecting body fluids, which includes a vacuum PDMS chamber openly connected with a hollow microstructure having a polymer cap. FIG. 5b shows an embodiment of a device for collecting body fluids. FIG. 5c shows one embodiment showing the principle of blood collection using a device for collecting body fluids. FIG. 5d shows a state in which a polymer cap formed on the tip of a microstructure is removed.

[0046] FIG. 6 illustrates formation of a hollow barrier by a dipping method. FIG. 6a shows the procedure of the dipping method for forming a polymer coating, and FIG. 6b is a graph showing an increase in the thickness of a polymer coating according to the number of a dipping process.

[0047] FIG. 7 illustrates formation of a hollow barrier by a capping method. FIG. 7a shows the procedure of the capping method for forming a polymer cap, FIG. 7b is a schematic view of an exposed-tip capping structure, and FIG. 7c shows one embodiment of the exposed-tip capping structure and an embedded-tip capping structure, which are formed by the capping method. FIG. 7d shows a result of measuring penetration force of a device in which a polymer cap having an exposed-tip or embedded-tip capping structure is formed. FIG. 7e is a schematic view of the embedded-tip capping structure.

[0048] FIG. 8 shows embodiments of polymer caps. FIG. 8a shows embodiments of polymer caps manufactured using carboxymethyl cellulose (CMC) and the uses thereof, and FIG. 8b shows embodiments of polymer caps manufactured using hyaluronic acid (HA) and the uses thereof.

[0049] FIG. 9 is a schematic diagram showing a pre-vacuum activation and a parylene film deposition process of a device for collecting body fluids. FIG. 9a is a cross-sectional view illustrating vacuum and airproof treatments of a device for collecting body fluids. FIG. 9b shows the fluid receiving part outer wall before parylene coating, and FIG. 9c shows the fluid receiving part outer wall after parylene coating.

[0050] FIG. 10 shows an embodiment of the use of a device for collecting body fluids in a simulated body fluid model. FIG. 10a shows an embodiment of the use of the device in which a hollow is blocked by a polymer coating, and FIG. 10b shows an embodiment of the use of the device in which a hollow is blocked by a polymer cap.

[0051] FIG. 11 shows the results of experiments on the relationship between a fluid capacity and the volume of a fluid receiving space.

MODES OF THE INVENTION

[0052] Hereinafter, the present invention will be described in detail by describing exemplary embodiments of the invention. It will be obvious to those skilled in the art that these embodiments are only provided to more particularly explain the present invention and the scope of the present invention is not limited thereto.

EXAMPLES

Example 1: Manufacture of Device for Collecting Fluids

[0053] A painless hollow microstructure with an optimized structure for minimally collecting invasive body fluid was manufactured by a drawing lithography-based technology based on conventional technologies [29, 30]. A polydimethylsiloxane (PDMS) chamber was prepared by pouring a 10:1 (v/v) prepolymer mixture of Sylgard 184 elastomer and a fining agent (manufactured by Dow Corning) onto an aluminum master. The mixture was cured in an 80° C. oven for one hour, and the resulting PDMS mold was carefully peeled off from the master. Subsequently, the hollow microstructures were assembled in a concentric shaft, thereby completing a one-touch body fluid collection instrument not having a polymer cap. Three aluminum masters were used to manufacture PDMS chambers having different volumes. The internal heights of the cylindrical chambers were 4 mm, and the internal diameters thereof were 3, 4, and 5 mm, respectively. Three PDMS chambers having different volumes of 28.26, 50.24, and 78.50 μl were manufactured.

Example 2: Manufacture of Hollow Microstructure, Hollow of which is Blocked by Polymer Coating or Polymer Cap

[0054] 2-1. Polymer Solution

[0055] Polyvinylpyrrolidone (PVP, 36 kDa, Sigma), carboxymethylcellulose (CMC, 90 kDa, low-viscosity, Sigma), and sodium hyaluronic acid (HA, 29 kDa, Soliance) were used as substrate polymers to block a hollow structure of a tip of a microstructure by dipping and capping methods. A PVP powder was dissolved in each of green and red dye solution (1%, w/v) at room temperature to prepare green and red dyes (Tartrazine, Bowon) containing a viscous PVP (35%, w/v) solution. CMC and HA powders were respectively dissolved in pink and yellow dye solutions (1%, w/v), thereby preparing a pink dye (Tartrazine, Bowon) containing a CMC (10%, w/v) solution and a yellow dye containing a HA (30%, w/v) solution, respectively.

[0056] 2-2. Dipping Method

[0057] A hollow microstructure was fixed in the downward direction on a micropositioner (Syringe pump, New Era Pump Systems) used to control position and a dipping rate. The microstructure was vertically dipped at a rate of 50 mm/min into a reservoir containing a 100 μl polymer coating solution until a hollow structure of a tip of the microstructure was completely submerged in a viscous PVP (35%, w/v) solution. A speed used to collect microstructure from a coating solution was maintained at 5 mm/min so as to form a polymer film on a surface of the tip of the microstructure. Immediately after the collection step, the coated polymer was air-dried. This dip-coating procedure was repeated until a polymer coating 31 having a desired thickness was obtained.

[0058] 2-3. Capping Method

[0059] A single polymer droplet was prepared using a solution dispenser (ML-5000X, Musashi) and automated X, Y, and Z stages (SHOT mini 100-s, Musashi) by discharging previously prepared polymer solutions onto the surface of a PDMS base layer at 0.9 kgf/cm for 0.05 seconds. The prepared PDMS base layer with the dispensed polymer droplet was fixed in the downward direction on a micropositioner (syringe pump, New Era Pump Systems) and was in contact with a microneedle, which was placed below the micropositioner, until the sharp tip of the microneedle just passed through the center of the droplet and was inserted into the PDMS base layer at a rate of 5 mm/min at room temperature for 4 min, to solidify the polymer. Finally, an isolation step was performed to separate the microstructure, on which the polymer-capped were formed, from the PDMS base layer at a rate of 50 mm/min.

[0060] 2-4. Mechanical Strength Analysis

[0061] The mechanical force required for a tip-blocked microstructure to penetrate the skin was measured by a displacement-force analyzer (Z0.5TN, Zwick) using 0.4-mm-thick synthetic skin (Gilchrist et al. 2008, Polyurethane elastomers, John Burn Inc., UK), which simulated the high-resistance stratum corneum of human skin. The tip-blocked microstructure was attached to the sensor probe of the force analyzer and pressed against the synthetic skin sheet at a speed of 60 μm/s via penetration force application. The penetration force of the tip-blocked microstructure was determined by measuring the maximum force before the penetration. The force was recorded as a function of displacement associated with the microstructure.

Example 3: Pre-Vacuuming and Coating with Parylene

[0062] The Parylene-C dimer (Femto Science Co.) and the microprocessor-controlled parylene coating system (Femto Science Co.) are commonly used to perform surface treatment with parylene. A parylene-C film with a thickness of 1 μm was thermally deposited on the surface of the closed blood extraction system by the following polymerization steps: (1) evaporation: parylene-C dimers (di-para-xylene) were first vaporized at a temperature of 160° C., (2) pyrolysis: the dimers were pyrolyzed at a temperature of 650° C. to form a highly reactive monomer (para-xylene) of parylene-C, and (3) deposition: the resulting polymer (poly-para-xylene) was finally coated onto the blood extraction system surface at room temperature. All of these processes were performed under vacuum conditions of less than 0.1 torr for 1.5 hours. The thickness of the parylene-C film was controlled by setting a quartz crystal microbalance (QCM) in the deposition chamber, and the film thickness was calculated by monitoring a frequency change in the QCM during deposition. The QCM change was measured from the beginning of the evaporation step, and the deposition was finished when the QCM frequency shift reached the value corresponding to the target thickness of the parylene-C film.

Example 4: Characterization of Surface Properties

[0063] The surface morphologies of the vacuum PDMS chamber with parylene coating were characterized by scanning electron microscopy (SEM; JEOL-7001, JEOL Ltd.) at an accelerating voltage of 15 kV. All samples were coated with a thin platinum layer by means of a sputtering machine for 100 seconds to produce a conductive surface prior to observation by SEM.

Example 5: Operation of Device for Collecting Body Fluid

[0064] FIG. 1c illustrates an operation principle of an embodiment of the device for collecting body fluids of the present invention.

[0065] The top of the microstructure tip of the fluid collection part of the device for collecting body fluids of the present invention was attached to a fluid collection site of a subject and was applied with a vertical force greater than the penetrating force that could penetrate the skin barrier of the subject so that the fluid collection part 2 was inserted into the subject skin.

[0066] In the case of a device with the polymer coating 31, the hollow of the fluid collection part was opened after a lapse of a decomposition time of the polymer coating and thus pressure gradients were formed, whereby a fluid from a subject was collected into the fluid receiving space 12 of the fluid receiving part 1.

[0067] In the case of the device with the polymer cap 32, the fluid collection part was inserted into the subject skin and, at the same time, the polymer cap 32 was removed. Immediately thereafter, pressure gradients were formed inside and outside the fluid receiving part, and thus, fluid from the subject was collected into the fluid receiving space 12 of the fluid receiving part 1.

Example 6: In-Vitro Blood Extraction

[0068] The liquid extraction capability of the blood extraction device was evaluated using distilled water (DW) and human whole-blood samples treated with 8% (v/v) anticoagulant solution CPDA-1. Each system was attached to a pusher, which was connected to the micropositioner (syringe pump, New Era Pump Systems) to allow accurate control of the insertion process for liquid extraction. The device was driven at a rate of 50 mm/min to insert the polymer-capped microstructure into the sample container with a synthetic skin cover to perform one-touch liquid extraction. After sampling, the system was removed from the sample container, and the extraction volumes of the liquid samples were measured using a 100 μl syringe (Hamilton).

Example 7: Blood Extraction Amount Control According to Volume Control of Fluid Receiving Space 12 of Fluid Receiving Part 1

[0069] A blood collection experiment was performed by means of a device for collecting a fluid equipped with a single hollow microstructure. This experiment was performed using a hollow microstructure with a 15° bevel angle and an inner diameter of 60 μm and a PDMS chamber with an internal negative pressure having a liquid containment space of 28.26 μl, 50.24 μl, or 78.50 μl. Blood collection was performed by applying the one-touch blood collection device to the virtual skin reproduction chamber containing a blood sample.

[0070] Results

[0071] 1. Self-Powered One-Touch Body Fluid Extraction Device

[0072] As shown in FIGS. 1 and 5A, the self-powered one-touch body fluid extraction system consists of three parts: 1) a fluid receiving part for providing power for the blood extraction by pressure gradient-driven force and for storing the extracted blood sample; 2) a fluid collection part including a hollow microstructure, which was minimally invasive and optimized for the skin, to induce the micro-channel for blood sampling; and 3) a hollow barrier for selectively blocking the hollow structure of the tip of the microstructure to maintain a closed pre-vacuum system and for precisely controlling the initiation of the blood sampling process.

[0073] A PDMS chamber corresponding to a fluid receiving part was manufactured using a traditional microfabrication technique [27, 28] and was designed as a 7 mm×7 mm×5 mm cube, with a cylindrical chamber having a 4 mm height and a 4 mm inside diameter. The hollow microstructure with an inner diameter of 60 μm, an outer diameter of 130 μm, and a 15° bevel angle was fabricated by drawing lithography, and the use of this optimized microstructure for minimally invasive blood extraction was demonstrated in previous studies [29, 30]. A real microscopic image of this system is illustrated in FIG. 5b.

[0074] The operating principle of this system is illustrated in FIG. 5c. The one-touch press system induces insertion of the hollow microstructure 21 into the skin by inducing the separation of the polymer cap 32 from the tip of the microstructure under the resistance force of the skin barrier. Simultaneously, the blood sample was extracted into the PDMS chamber by the pressure gradient between the fluid receiving space 12 of the fluid receiving part, in which was formed, and the blood vessel. The transparent property of the PDMS layer facilitates observation of a process of separating the polymer cap from the tip of the microstructure, and thus, is was used to reproduce the barrier function of the skin. A functional separation principle of the polymer cap 32 is illustrated in FIG. 5d.

[0075] 2. Tip for Blocking Hollow Microstructure

[0076] In particular, biocompatible polymers PVP, CMC, and HA have been widely used as structural materials for the fabrication of coated and dissolved microstructures in medical applications based on micro-fabrication techniques such as a dip-coating method and drawing-lithograph [31, 32]. In the present invention, the optimal concentrations of such polymers were used to manufacture a tip-blocked hollow microstructure structure using dipping and capping methods. Subsequently, to induce pre-vacuum activation, as a subsequent process, for the closed one-touch blood extraction system, the tip-blocked microstructure was openly connected to the PDMS chamber.

[0077] 3. Polymer-Coated Hollow Microstructure

[0078] The hollow microstructure was coated with viscous PVP (35%, w/v) solution with red dye using a microscale dipping method developed previously [33, 34]. As shown in FIG. 6a, the dipping method consisted of two distinct steps occurring in sequence: i) a dipping step: controlling to dip the tip of the microstructure into the polymer solution to determine a coating area and ii) a withdrawal and drying step: withdrawing the tip of the microstructure from a polymer solution at a rate of 5 mm/min and coating a viscous PVP polymer on a surface of the microstructure. By the drying, a solid polymer coating was attached onto the tip of the microstructure

[0079] In manufacturing the polymer-coated microstructure, it is important to block the hollow structure of the tip of the microstructure and allow the polymer coating to attach to the tip of the microstructure without destruction during subsequent processes, such as pre-vacuum treatment, for example, by keeping an interior pressure of the vacuum chamber lower.

[0080] To accomplish this, microstructures having different polymer coatings were manufactured, connected to vacuum chambers, and subjected to experiments. The thicknesses of the polymer coatings were controlled by the number of dips; polymer coating thicknesses of 3.25±0.44 μm, 16.1±2.09 μm, 39.53±1.93 μm, and 78.6±12.26 μm were obtained by dipping the tip of the microstructure into the coating solution one, two, four, and six times, respectively. Results are illustrated in FIG. 6b. The thickness of the polymer coating on the microneedle tip increased as the number of dips increased. To guarantee physical strength to ensure block of the hollow structure under a lower pressure condition, a polymer coating thickness was determined to be at least 39.53±1.93 μm (dipping into the coating solution four times).

[0081] 4. Polymer-Capped Hollow Microstructure

[0082] The hollow microstructure was capped with a viscous PVP (35%, w/v) polymer solution with green dye using a novel micron-scale capping method suggested by the present inventors. As shown in FIG. 7a, this capping method consists of three steps. (i) First, a capping polymer solution is provided on the surface of a PDMS base layer to form a single polymer droplet. In particular, the height of the solid polymer cap on the tip of the microstructure can be controlled by adjusting the amount of the polymer in a droplet using a dispenser. As illustrated in FIG. 7b, it was required to provide a size higher than 204 μm (H=L (an inner diameter of hollow microstructure)/tan θ)=60/tan 15° so as to guarantee complete block of the needle. With increasing drying time of a polymer droplet, the height of the polymer droplet was decreased. However, after four minutes, the height of a solidified structure was not changed because evaporation of distilled water was completed after four minutes. Therefore, to form a solid polymer cap having a height of 250 μm, it is required to provide a polymer droplet having a height of at least 400 μm on a surface of the PDMS base layer. Since a surface of the PDMS base layer is hydrophobic, a contact angle of the PDMS base layer to the polymer solution may be decreased by increasing the height of the droplet, and the polymer solution may be easily isolated along with the solid polymer cap in the isolation step.

[0083] (ii) In a second step, a polymer droplet fixed in the downward direction was precisely controlled to contact with the tip of the microstructure. As illustrated in FIG. 7c, an embedded-tip (see FIG. 7e) or exposed-tip (see FIG. 7b) capping structure may be manufactured by controlling the position of the microstructure tip in the droplet. To form the tip of the microstructure with an embedded-tip capping structure, the microstructure tip was inserted into the middle of a polymer droplet without insertion into a PDMS base layer. On the other hand, to form an exposed-tip capping structure, the top of the tip of the microstructure was inserted into a smooth PDMS base layer via a droplet. To visually control the position of the microstructure tip inside the PDMS base layer, the transparent property of PDMS was used. (iii) Finally, to solidify the polymer, the microstructure was allowed to stand at room temperature for four minutes and then a separation process of separating the polymer-capped microstructure from the PDMS based layer was performed.

[0084] To accomplish successful blood extraction by means of a microstructure, the microstructure should have strength sufficient to penetrate the skin barrier without damage. Both the blocking microstructures by capping exhibit penetration force smaller than an average fracture force (5.85±0.25 N) of hollow microstructures. Penetration forces are summarized in Table 1.

TABLE-US-00001 TABLE 1 Penetration force (N) Displacement (mm) No blocking 0.78 ± 0.01 1.47 ± 0.03 Blocking by capping (exposed tip) 1.31 ± 0.01 1.63 ± 0.02 Blocking by capping (embedded 2.30 ± 0.10 2.13 ± 0.05 tip)

[0085] Using a micron-scale capping method, the hollow microstructures were respectively capped with viscous CMC (10%, w/v) and HA (30%, w/v) polymer solutions respectively including pink and yellow dyes. A polymer cap separation process is illustrated in FIG. 8.

[0086] 5. Pre-Vacuum Activation and Parylene Film Deposition

[0087] The high gas permeability properties of PDMS were used to induce inner pre-vacuum activation for the closed system. On the other hand, the gas leaking through the pores of PDMS made it difficult to maintain the applied negative pressure. Parylene is widely used as an inert coating material to enhance biomedical compatibility due to its low permeability to gases and moisture, and may be coated on any geometric surfaces even having holes and cracks [35, 36]. Therefore, to form a highly closed system, the pre-vacuum activation of the one-touch blood extraction system may be maintained over a long period of time (more than 10 hours) by allowing the parylene deposition to effectively seal the porous structure of the PDMS chamber. The contents were confirmed by the Satoshi Konisni group [37]. All of the procedures for vacuumizing and airproofing were performed by a single parylene film deposition process.

[0088] First, all of the closed blood extraction systems were formed by blocking the tip of the microstructure with the polymer using the capping methods. Subsequently, pre-vacuum activation was performed by maintaining a low-pressure condition for at least 20 minutes. The low-pressure condition was established before a parylene film deposition process, and a low vacuum condition of less than 0.1 torr was maintained for 1.5 hours. FIG. 9a schematically illustrates the pre-vacuum activation and parylene film deposition process. Air molecules form vacuum through the porous structure of the PDMS to form a vacuum chamber. Subsequently, parylene was deposited on the surface of PDMS to successfully seal the porous structure with parylene caulking inside and with 1 μm-thick parylene coating outside. FIGS. 9b and c illustrate the scanning electron microscope (SEM) images of the PDMS chamber surface before and after the deposition with parylene and 1 μm-thick parylene film coating. Potential energy was stored inside the pre-vacuum activated blood extraction system which has been airproofed, thereby providing the driving force for blood sampling when interconnected with a blood vessel via the channel of the hollow microneedle. The pre-vacuum activation mechanism demonstrated that this system can be used for blood sampling without the need for an external power source and, further, the volume of an extracted sample may be adjusted by controlling the volume of the vacuum chamber.

[0089] 6. Fluid Extraction Control

[0090] Press force was applied to the blood extraction system to induce insertion of the microstructure into the sample container by passing through the synthetic skin cover for the liquid extract. It was observed that the polymer coated on the tip of the microstructure was inserted into the container with the microstructure without residues on a surface of the synthetic skin by the dipping method. Accordingly, to dissolve the polymer coated on the tip of the microstructure, sample extraction was initiated after 120 seconds of insertion of the microstructure into the sample container and this extraction process was completed within four seconds. However, the capping polymer was removed for the surface of the microstructure tip and moved in a downward direction due to resistance of a synthetic skin. As illustrated in FIG. 10, extraction was immediately performed within four seconds.

[0091] 7. Blood Extraction Volume Control

[0092] Blood extraction was performed using a device equipped with each of vacuum PDMS chambers having different volumes of 28.26 μl, 50.24 μl, and 78.50 μl. As results, blood samples were respectively obtained in total volumes of 14.3 μl, 28.3 μl, and 43.7 μl sufficient for microsystem analysis within five seconds (see FIG. 11).

[0093] Although some embodiments of the present invention have been described in detail, it is obvious to those skilled in the art that the embodiments are merely provided as preferred examples and the scope of the present invention is not limited thereto. Therefore, the substantial range of the present invention is defined by the appended claims and equivalent thereof.

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