Mucosal cell collection device and mucosal cell collection method

Abstract

This mucosal cell collection device 1 comprises: an ultrasound radiation means 11; a tubular member 13 which is provided around the ultrasound radiation means 11 and which has a protrusion 12 that protrudes from one end of the ultrasound radiation means 11; an introduction means 16 by which a liquid is introduced into a space 15 formed by the ultrasound radiation means 11, the protrusion 12 of the tubular member 13 and a specimen 14; and an extraction means 17 by which cells 14b that have separated from a mucous membrane 14a of the specimen 14 are extracted together with the liquid from the space 15.

Description

Technical Field

[0001] The present invention relates to a mucosal cell collection device and a mucosal cell collection method. The present invention claims priority based on Japanese Patent Application No. 2023-008199 filed in Japan on January 23, 2023, the content of which is incorporated herein by reference.

Background Art

[0002] According to survey results, approximately 21% of the general adult population in Japan suffers from insomnia due to sleep disorders, and approximately 15% of the general adult population in Japan is aware of daytime sleepiness. Long-term sustained sleep deprivation and sleep disorders may increase the susceptibility to lifestyle-related diseases and depression. It is important to appropriately address sleep disorders. Sleep disorders are considered to be caused by disruption of the body clock (circadian rhythm). Understanding circadian rhythm is extremely useful in the diagnosis and treatment of sleep disorders. It is known that the expression level of clock genes that govern circadian rhythm increases and decreases in accordance with the circadian rhythm cycle. By measuring the expression level of clock genes in a sample and comparing it with a molecular timetable, it is possible to accurately know what time of the internal body clock it is at the time when the sample is collected. That is, to grasp a person's circadian rhythm, it is necessary to collect cells every few hours during daily life and measure the expression level of clock genes. For this measurement, a device that can appropriately collect cells during daily life is useful. In addition, appropriately measuring the expression level of proteins useful for diagnosis and disease condition evaluation by collecting cells is expected to lead to unprecedented inspection and diagnosis. The oral cavity is sometimes called a mirror that reflects an individual's health status. Changes indicating disease appear as changes in the oral mucosa, which may reveal systemic conditions such as diabetes and vitamin deficiency, or local effects caused by chronic intake of tobacco and alcohol.

[0003] Patent Document 1 describes a liquefaction collection device for collecting cells from mucosal tissue without destroying the cell membrane, wherein the reservoir housing contains an ultrasonic generator and a liquefaction-promoting medium LPM (such as PBS) that transmits ultrasonic energy to the mucosal tissue and converts tissue components into a soluble state, the reservoir housing forms a closed space in which the ultrasonic generator, LPM and tissue are in contact, the LPM is introduced into the space from a spray system device via a tube, and the liquefied tissue sample is collected from the reservoir housing via a tube into a sample container by the pressure of a continuously applied suction pump.

[0004] Patent Document 2 describes a probe, which is a tissue collection device for a diagnostic system that collects tumor markers in prostate tissue, comprising a cylindrical cartridge that contacts the tissue, and having an ultrasonic transducer and an aluminum rod that transmits its energy, and a liquid vibrating coupling medium that fills the space between the tip of the rod and the tissue within a chamber which is the internal space of the cartridge, wherein the medium generates cavitation to effectively transmit ultrasonic energy to the tissue and collect epithelial cells.

[0005] Patent Document 3 describes a Helicobacter pylori collection system in which fluid is injected into the stomach from outside the body through an injection channel formed in the insertion part of an endoscope introduced into the human body, ultrasonic vibrations are applied to the gastric wall via the injected fluid using an ultrasonic transducer introduced into the stomach through the lumen of the endoscope, the fluid in the stomach is aspirated using a suction device, and Helicobacter pylori is filtered and collected using a bacterial collection filter on the suction tube.

[0006] Non-patent document 1 describes a portable STAMP device (Surfactant-based Tissue Acquisition for Molecular Profiling) that collects nucleic acids, proteins, etc., in the same ratio as in living organisms, by equipping the internal space of a cylindrical housing that comes into contact with tissue with one side of an ultrasonic element and a sampling buffer. [Prior art documents] [Patent Documents]

[0007] [Patent Document 1] Japan Special Publication No. 2012-518171 [Patent Document 2] U.S. Patent No. 6589173 [Patent Document 3] Japanese Patent Publication No. 2001-245890 [Non-patent literature]

[0008] [Non-Patent Document 1] Sumit Paliwal, et al., “One-step acquisition of functional biomolecules from tissues,” Proceedings of the National Academy of Sciences, August 17, 2010, Vol. 107, No. 33, pp. 14627-14632. [Overview of the initiative] [Problems that the invention aims to solve]

[0009] Cell collection is generally performed using blood or oral mucosa. Cell collection from blood is highly invasive because it involves puncturing blood vessels with a needle, making it difficult to perform multiple cell collections in a single day. Cell collection from oral mucosa involves scraping cells from the mucosa, which can lead to cell damage that interferes with analysis, or contamination by substances from the oral cavity that can interfere with analysis.

[0010] This invention has been made in view of the above circumstances, and aims to provide a minimally invasive mucosal cell collection device and mucosal cell collection method. [Means for solving the problem]

[0011] The inventors of the present invention conducted extensive research to achieve the above objective and discovered that mucosal cells can be collected in a minimally invasive manner by irradiating the mucous membrane of a subject with ultrasound through a liquid and extracting the cells separated from the mucous membrane of the subject together with the liquid, thus completing the present invention. The present invention includes the following embodiments.

[0012] [1] Ultrasonic irradiation means, A cylindrical member provided around the ultrasonic irradiation means and having a protruding portion that extends from one end of the ultrasonic irradiation means, An introduction means for introducing liquid into the space formed by the ultrasonic irradiation means, the protruding portion of the cylindrical member, and the subject, and Extraction means for removing cells separated from the mucous membrane of the subject along with the liquid from the space. A mucosal cell collection device equipped with the following features. [2] The mucosal cell collection apparatus according to [1], wherein the ultrasonic irradiation means comprises an ultrasonic transducer and an ultrasonic transmission member that transmits ultrasonic waves generated by the ultrasonic transducer. [3] The mucosal cell collection apparatus according to [1] or [2], wherein the introduction means and the extraction means each comprise a tube. [4] The mucosal cell collection device according to [1] or [2], further comprising a sealing member between the ultrasonic irradiation means and the cylindrical member. [5] The mucosal cell collection device according to [2], wherein the ultrasonic transducer includes a disc portion formed in the shape of a disc and is configured to vibrate laterally along the radial direction of the disc portion. [6] The mucosal cell collection device according to [5], wherein the ultrasonic transmission member converts the vibration of the ultrasonic transducer into vibration in a vertical direction perpendicular to the lateral direction. [7] The mucosal cell collection device according to [6], wherein the ultrasonic transmission member is formed in the shape of a cymbal. [8] The mucosal cell collection device according to [7], wherein the ultrasonic transmitting member comprises an outer peripheral portion connected to the disc portion, an inclined portion connected to the outer peripheral portion and extending inclined away from the disc portion, and a convex portion connected to the inclined portion and convex in the opposite direction to the disc portion. [9] The mucosal cell collecting device according to [8], wherein the cylindrical member is provided around the convex portion.

[10] The mucosal cell collecting device according to [9], wherein the maximum lateral dimension is 11 mm or less, and the maximum longitudinal dimension is 10 mm or less.

[11] The mucosal cell collecting device according to [2], wherein the ultrasonic transmission member is formed into a shape in which a cross-sectional area orthogonal to the axial direction of the ultrasonic transmission member decreases toward the tip of the protruding portion.

[12] The mucosal cell collecting device according to

[11] , wherein the cylindrical member is formed into a shape in which a cross-sectional area orthogonal to the axial direction of the cylindrical member decreases toward the tip of the protruding portion.

[13] The mucosal cell collecting device according to [2], wherein the cylindrical member is attached to the ultrasonic transmission member at a position corresponding to a node when the ultrasonic transmission member vibrates due to ultrasonic waves generated by the ultrasonic transducer.

[14] The mucosal cell collecting device according to

[13] , further comprising a seal member between the ultrasonic transmission member and the cylindrical member, wherein the seal member is attached to the ultrasonic transmission member and / or the cylindrical member at the position corresponding to the node.

[15] an ultrasonic irradiation means, a cylindrical member provided around the ultrasonic irradiation means and having a protruding portion protruding from one end of the ultrasonic irradiation means, an introducing means for introducing a liquid into a space formed by the ultrasonic irradiation means, the protruding portion of the cylindrical member and a subject, and an extracting means for extracting cells separated from the mucosa of the subject together with the liquid out of the space A mucosal cell collecting method using a mucosal cell collecting device, comprising: a step of irradiating the mucosa of the subject with ultrasonic waves via the liquid, and a step of extracting cells separated from the mucosa of the subject together with the liquid A mucosal cell collecting method, comprising:

[16] The method for collecting mucosal cells according to

[15] , further comprising, before the irradiating step or at an initial stage of the irradiating step, filling the space formed by the ultrasonic wave irradiation means, the protruding portion of the cylindrical member and the subject with the liquid, circulating the liquid, and washing and removing oral substances in the space and / or oral substances adhering to a mucosal surface of the subject with the liquid. [Effects of the Invention]

[0013] According to the present invention, a minimally invasive mucosal cell collection device and a minimally invasive mucosal cell collection method can be provided. Furthermore, according to the present invention, there is no need to scrape cells from the oral mucosa, and cells are less likely to be destroyed, so that analysis is not hindered. Furthermore, according to the present invention, since the mucosal surface is washed before cell collection, oral substances that hinder analysis are less likely to mix into collected mucosal cells, and thus analysis is not hindered. [Brief Description of the Drawings]

[0014] [Figure 1] Fig. 1 is a longitudinal cross-sectional view of a mucosal cell collection device according to an embodiment of the present invention. [Figure 2] Fig. 2 is an end view seen from the lower end of the mucosal cell collection device in Fig. 1. [Figure 3] Fig. 3 is a diagram for explaining a mucosal cell collection method according to an embodiment of the present invention. [Figure 4] Fig. 4 is a graph showing measurement results of Experimental Example 1. A horizontal axis of the graph represents frequency [kHz], and a vertical axis of the graph represents ultrasonic intensity [W / cm2]. [Figure 5] Fig. 5 is a graph showing measurement results of Experimental Example 2. A horizontal axis of the graph represents applied voltage [Vp-p], and a vertical axis of the graph represents ultrasonic intensity [W / cm2]. [Figure 6] Fig. 6 is a photograph showing an outline of an experimental operation of Experimental Example 3. [Figure 7] Fig. 7 is a graph showing an outline of an experimental operation of Experimental Example 3. [Figure 8] Fig. 8 is a photograph (fluorescence microscope, eyepiece) showing the results of Experimental Example 3. [Figure 9] Figure 9 is a photograph (fluorescence microscope scan) showing the results of Experimental Example 3. [Figure 10] Figure 10 shows the experimental schedule for Experiment Example 4. [Figure 11] Figure 11 shows the experimental schedule for Experiment Example 4. [Figure 12] Figure 12 shows the experimental schedule for Experiment Example 4. [Figure 13] Figure 13 is a graph showing the results of Experimental Example 4. The horizontal axis of the graph represents the applied voltage [Vp-p], and the vertical axis represents the number of cells collected. [Figure 14] Figure 14 shows the experimental schedule for Experiment Example 6. [Figure 15] Figure 15 shows the experimental schedule for Experiment Example 6. [Figure 16] Figure 16 shows the experimental schedule for Experiment Example 6. [Figure 17] Figure 17 is a graph showing the results of Experimental Example 6. The horizontal axis of the graph represents the sampling time [min], and the vertical axis represents the number of cells collected. [Figure 18] Figure 18 shows the experimental schedule for Experiment Example 7. [Figure 19] Figure 19 shows the experimental schedule for Experiment Example 7. [Figure 20] Figure 20 is a graph showing the results of Experiment Example 7. The horizontal axis of the graph represents the immersion time [min], and the vertical axis represents the number of cells collected. [Figure 21] Figure 21 shows the experimental schedule for Experiment Example 8. [Figure 22] Figure 22 is a graph showing the results of Experiment Example 8. The horizontal axis of the graph represents the number of samplings, and the vertical axis represents the number of cells collected. [Figure 23] Figure 23 is a perspective view of the mucosal cell collection device according to the second embodiment. [Figure 24] Figure 24 is a perspective view showing an example in which an endoscope is applied to the mucosal cell collection device according to the second embodiment. [Figure 25]Figure 25 is a longitudinal cross-sectional view of a mucosal cell collection device according to the second embodiment. [Figure 26] Figure 26 is a diagram illustrating the conversion of the vibration direction by the ultrasonic transmission member according to the second embodiment. [Figure 27] Figure 27 is a longitudinal cross-sectional view of a mucosal cell collection device according to the third embodiment. [Figure 28] Figure 28 is an end view of the mucosal cell collection device shown in Figure 27, viewed from the bottom. [Figure 29] Figure 29 is a diagram illustrating the mounting position of the cylindrical member and / or sealing member according to the third embodiment. [Figure 30] Figure 30 shows the mounting positions of the sealing member, from position (1) to position (4). [Figure 31] Figure 31 shows the relationship between the mounting position of the sealing member and the relative value of the output ultrasonic intensity. [Modes for carrying out the invention]

[0015] In this specification and the claims, the "~" indicating a numerical range means that the numbers before and after it are included as the lower and upper limits, respectively.

[0016] The embodiments of the present invention will be described below with reference to the drawings as appropriate, but the present invention is not limited to the embodiments and drawings described below, and various modifications are possible as long as they do not depart from the spirit of the invention.

[0017] [Mucosal cell collection device] The mucosal cell collection apparatus of the present invention comprises an ultrasonic irradiation means, a cylindrical member provided around the ultrasonic irradiation means and having a protruding portion that extends from one end of the ultrasonic irradiation means, an introduction means for introducing liquid into the space formed by the ultrasonic irradiation means, the protruding portion of the cylindrical member, and the subject, and an extraction means for extracting cells separated from the mucosa of the subject together with the liquid from the space. The mucosal cell collection device of the present invention will be described below with reference to one embodiment.

[0018] The mucosal cell collection device 1, shown in Figure 1 in longitudinal section, comprises an ultrasonic irradiation means 11, a cylindrical member 13 provided around the ultrasonic irradiation means 11 and having a protrusion 12 protruding from one end of the ultrasonic irradiation means 11, an introduction means 16 for introducing liquid into the space 15 formed by the ultrasonic irradiation means 11, the protrusion 12 of the cylindrical member 13 and the subject 14, and an extraction means 17 for extracting cells separated from the mucosa 14a of the subject 14 together with the liquid from the space 15.

[0019] The ultrasonic irradiation means 11 consists of, for example, an ultrasonic transducer 11a and an ultrasonic transmission member 11b that transmits ultrasonic waves generated by the ultrasonic transducer 11a. The ultrasonic transducer 11a is a component that converts high-frequency power from the oscillator into ultrasonic vibrations. The ultrasonic transducer 11a is, for example, an electrostrictive or magnetostrictive type. The electrostrictive type expands and contracts when a voltage is applied, and the magnetostrictive type expands and contracts when a magnetic field is applied, thereby generating ultrasonic waves. As the ultrasonic transducer 11a, for example, a Langevin transducer, which is an electrostrictive ultrasonic transducer, is preferred. The resonant frequency of the ultrasonic transducer 11a is not particularly limited, but 20 to 100 kHz is preferred. The ultrasonic transmission member 11b is preferably made of a metal that has excellent transmission efficiency for ultrasonic waves generated by the ultrasonic transducer 11a. Examples of metals include iron alloys such as steel, stainless steel, maraging steel, 42 alloy, Invar, Kovar, Sendust, Permendür, silicon steel, KS steel, and Spiegel Eisen; alloy steels such as Krupp steel, chromium molybdenum steel, manganese molybdenum steel, and Yasugi steel; copper alloys such as brass, red copper, Rombuck, nickel silver, bronze, cupronickel, red copper, Constantan, Nordic gold, and Knifé; aluminum alloys such as duralumin, Silmin, Hastelloy, Monel, Inconel, nichrome, Sunplatinum, and Permalloy; and magnesium alloys. The size of the ultrasonic irradiation means 11 can be appropriately set according to the site from which mucosal cells are collected. When used for collecting oral mucosal cells, the size of the ultrasonic irradiation means 11, if cylindrical, is preferably 0.05 to 0.2 m in length and 0.03 m or less in diameter.

[0020] The shape of the tubular member 13 is not particularly limited as long as it is cylindrical, but a cylindrical shape is preferred. Since the tubular member 13 is disposable after each cell collection, synthetic resin is preferred as the material of the tubular member 13. The tubular member 13 can be manufactured by various methods such as injection molding and 3D printing. The cylindrical member 13 is a separate component from the ultrasonic irradiation means 11. Preferably, the inner surface of the cylindrical member 13 is provided with a sealing member to prevent liquid leakage from the space 15. An O-ring 18 is preferred as the sealing member, and a groove 13a for mounting the O-ring 18 is provided on the inner surface of the cylindrical member 13. The O-ring 18 is preferably elastically deformable and can be made of, for example, natural rubber, synthetic rubber, or a thermoplastic elastomer. The sealing member may be integrally molded with the cylindrical member 13. The gap between the inner surface of the cylindrical member 13 and the ultrasonic irradiation means 11 is not particularly limited, as long as the cylindrical member 13 can be attached to and detached from the ultrasonic irradiation means 11. The cylindrical member 13 is provided with a through hole 13b for connecting the space 15 and the introduction means 16, and a through hole 13c for connecting the space 15 and the extraction means 17. When the cylindrical member 13 and at least a part of the introduction means 16 and / or extraction means 17 are formed integrally, the lumen of the introduction means 16 and the through hole 13b and / or the lumen of the extraction means 17 and the through hole 13c become one. The tubular member 13 of the mucosal cell collection device 1 is preferably disposable, while the ultrasonic irradiation means 11 is preferably reusable. In particular, the tubular member 13 is preferably disposable because it can be manufactured inexpensively and contamination can be avoided.

[0021] The introduction means 16 includes, for example, a tube. The introduction means 16 is connected to a reservoir (not shown) and a pump (not shown) for supplying liquid to be introduced into the space 15. The extraction means 17 includes, for example, a tube. The extraction means 17 is connected to a recovery tank (not shown) for recovering the liquid extracted from the space 15.

[0022] The end view shown in Figure 2 is an end view of the mucosal cell collection device 1 as seen from the lower end. The cylindrical member 13, through hole 13b, through hole 13c, and O-ring 18 are the same as in Figure 1. The groove 13a of the cylindrical member 13 cannot be directly viewed, but the O-ring 18 is fitted into it. If the cylindrical member 13 is cylindrical in shape, the space 15 will be a roughly cylindrical space, but a slight protrusion is provided for the introduction and extraction of liquid through the through holes 13b and 13c. The outer diameter D1 of the cylindrical member 13, the diameter d1 of the substantially cylindrical space 15, and the diameter d2 of the portion including the groove 13a (= d1 + depth of groove 13a × 2) are not particularly limited. Also, the diameter d3 of the through hole 13b and the diameter d4 of the through hole 13c are not particularly limited.

[0023] Subject 14 is preferably a subject derived from a mammal such as a human. Mucosa 14a is the tissue that is always moist with mucus, forming the inner walls of organs such as the digestive tract, respiratory tract, excretory tract, and reproductive tract. Mucosa 14a is an epithelial layer derived from the ectoderm, covered with epithelial cells, and is involved in absorption and secretion. Mucosa 14a is located in various body cavities and faces the external environment and internal organs. Mucosa 14a connects to the skin in many places, such as the nostrils, lips, ears, reproductive tract, and anus. Mucosal epithelium is the epithelium that protects the surface of the mucosa. Therefore, in areas with strong mechanical stimuli (oral cavity, esophagus, anus, etc.), it is composed of "stratified squamous epithelium," while in areas where secretion and absorption occur (stomach, intestines, etc.), it is composed of "simple columnar epithelium." The lamina propria is a layer of connective tissue densely composed of collagen fibers. The muscularis mucosa is a thin layer of smooth muscle located below the lamina propria, separating the lamina propria from the submucosa. Examples of mucous membranes 14a include the buccal mucosa, gastric mucosa, intestinal mucosa, olfactory epithelium, oral mucosa, and endometrium. Oral mucosa and buccal mucosa are preferred because cell collection is easy using the mucosal cell collection device of this embodiment. Gastric mucosa, intestinal mucosa, and endometrium are also preferred if the mucosal cell collection device of this embodiment can be brought into contact with the mucosa via the forceps channel of an endoscope. The endometrium is the mucosal tissue that lines the inside of the uterus. In addition, the mucosal cell collection device of this embodiment can be applied to cell collection from serous membranes on the surface of organs in the abdominal cavity, thoracic cavity, etc., as well as to the pia mater in the ventricles of the brain. The serous membrane is a membrane that covers the surface of body cavities and the organs inside them and secretes serous fluid. The pia mater is the innermost membrane of the meninges that surround the brain and spinal cord, and is a thin, reticular membrane that completely covers the surface of the brain, including the ventricular surface.

[0024] Examples of the aforementioned liquids include buffer solutions that cause minimal damage to cells, such as phosphate-buffered saline (PBS).

[0025] [Mucosal cell collection method] The method for collecting mucosal cells according to this embodiment comprises the steps of irradiating the mucosa of a subject with ultrasound through a liquid, and extracting the cells separated from the mucosa of the subject together with the liquid.

[0026] In the mucosal cell collection method of this embodiment, as shown in Figure 3, the mucosal cell collection device 1 is pressed against the mucosa 14a of the subject 14, liquid is introduced into the space 15 via the introduction means 16, and ultrasonic waves generated by the ultrasonic irradiation means 11 are irradiated onto the mucosa 14a of the subject 14 through the liquid. As a result, cells 14b are separated from the mucosa 14a and dispersed in the liquid.

[0027] The mucosal cell collection method of this embodiment may further include a step of filling the space 15 formed by the ultrasonic irradiation means 11, the protruding portion 12 of the cylindrical member 13, and the subject 14 with the liquid and circulating it, before or at the beginning of the irradiation step, in which oral cavity substances in the space 15 and / or oral cavity substances attached to the surface of the mucosa 14a of the subject 14 are washed and removed with the liquid. Here, "initial stage of the irradiation process" refers to the first half of the total irradiation process time. By incorporating this process, it becomes possible to collect cells with fewer impurities.

[0028] When the mucosal cell collection device 1 is pressed against the mucosa 14a of the subject 14, the mucosal cell collection device 1 may be adsorbed to the mucosa 14a by creating negative pressure in the space 15.

[0029] By flowing the liquid through space 15 without irradiating it with ultrasound, substances present in space 15 that would interfere with the analysis can be washed away and removed.

[0030] Cells can be collected from the mucous membrane by removing the liquid in which the cells 14b are dispersed via the extraction means 17.

[0031] The tubular member 13 of the mucosal cell collection device 1 is preferably disposable, while the ultrasonic irradiation means 11 is preferably reusable. In particular, the tubular member 13 is preferably disposable because it can be manufactured inexpensively and contamination can be avoided. [Examples]

[0032] The present invention will be described in more detail below with reference to examples, but the present invention is not limited to the examples described later.

[0033] [Experimental Example 1] The frequency characteristics of the mucosal cell collection device 1 shown in Figure 1 were evaluated. A Langevin transducer (HEC-1540P2BF, manufactured by Honda Electronics Co., Ltd.; resonant frequency 40kHz) was used as the ultrasonic transducer 11a, and a cylindrical metal material with a diameter of 14 mm, made by machining brass, was used as the ultrasonic transmission member 11b. The outer diameter of the cylindrical member 13 was 21 mm, the diameter of the space 15 was 15 mm, and the diameter of the O-ring 18 was 17.1 mm. The space 15 was filled with water so that the protruding part 12 of the mucosal cell collection device 1 was submerged in water. A hydrophone (TC4013-1) was placed 1 mm from the lower end of the protruding part 12, and an ultrasonic transducer 11a was subjected to a voltage of 20 Vp-p to generate ultrasound. The measurement results are shown in the graph in Figure 4. In the graph in Figure 4, the horizontal axis represents frequency [kHz], and the vertical axis represents ultrasonic intensity [W / cm]. 2 ]

[0034] [Experimental Example 2] Using the same configuration as in Experimental Example 1, the applied voltage was changed and the ultrasonic intensity was measured. The results are shown in the graph in Figure 5. The frequency of the ultrasonic transducer 11a was set to 35.1 kHz. In the graph shown in Figure 5, the horizontal axis represents the applied voltage [Vp-p], and the vertical axis represents the ultrasonic intensity [W / cm²]. 2 ]

[0035] [Experimental Example 3] A portion of the esophagus was removed from a pig (Figure 6, upper left). The excised esophagus was cut and unfolded (Figure 6, upper right). The surface was washed (Figure 6, bottom left). Cell collection experiments were conducted from excised esophageal mucosa samples from pigs using the same mucosal cell collection device as in Experimental Example 1. Phosphate-buffered saline (PBS) was used as the liquid for cell collection. The applied voltage to the ultrasonic transducer was set to 20 Vp-p, and the frequency to 35.1 kHz. This is within the range where cavitation occurs. As shown in Figure 7, ultrasonic irradiation was started while injecting PBS (0 min), aspiration of PBS was started at a rate of 20 mL / h (5 min), and ultrasonic irradiation and injection / aspiration were terminated (20 min). The PBS containing the cells isolated from the mucous membrane was collected, placed in a centrifuge tube, and the cells were precipitated by centrifugation. The obtained cells were stained using a staining kit (Live / Dead Cell Staining Kit II, PromoKine). Figure 8 shows the results of observing stained cells through the eyepiece using a fluorescence microscope (IX81, Olympus Corporation) with excitation fluorescence at a wavelength of 488 nm, while Figure 9 shows the results of scanning cells from the fluorescence microscope. Based on the shape and size of the cells, it was determined that there was a high probability that epithelial cells had been successfully collected.

[0036] [Experimental Example 4] The experiment was conducted in the same manner as in Experiment Example 3, except that the ultrasonic intensity was changed as shown in Figures 10 to 12. The results are shown in the graph in Figure 13. In the graph in Figure 13, the horizontal axis represents the applied voltage [Vp-p], and the vertical axis represents the number of cells collected. The higher the applied voltage (stronger the ultrasonic intensity), the more cells could be collected.

[0037] [Experimental Example 5] (Comparative Example) The experiment was conducted in the same manner as in Experiment Example 3, except that ultrasound irradiation was omitted. Only eight cells were collected.

[0038] [Experimental Example 6] The experiment was conducted in the same manner as in Experiment Example 3, except that the cell collection time was changed as shown in Figures 14 to 16. The results are shown in the graph in Figure 17. In the graph in Figure 17, the horizontal axis represents the sampling time [min], and the vertical axis represents the number of cells collected. The longer the sampling time, the more cells could be collected.

[0039] [Experimental Example 7] (Comparative Example) To confirm the difference in efficiency between a mucosal cell collection device and the common method of collecting cells from mucosal tissue using a cotton swab, we conducted a cell collection experiment by swabbing the mucosa with a cotton swab. The experimental schedule is shown in Figures 18 and 19. The results are shown in the graph in Figure 20. In the graph in Figure 20, the horizontal axis represents the immersion time [min], and the vertical axis represents the number of cells collected. While longer immersion times allowed for the collection of more cells, the number was still less than when using a mucosal cell collection device. Furthermore, cells collected using cotton swabs were mostly smaller than 14 μm, suggesting contamination with impurities or fragmented cells. Therefore, genetic analysis was expected to be difficult.

[0040] [Experimental Example 8] Since we plan to collect cells every four hours, or six times a day, to measure the dynamic changes in RNA transcription or protein expression from genes, the purpose is to confirm what effect six cell collections from the same site have on the tissue. The experiment was conducted according to the experimental schedule shown in Figure 21. The results are shown in the graph in Figure 22. In the graph in Figure 22, the horizontal axis represents the number of samplings, and the vertical axis represents the number of cells collected.

[0041] Approximately 3,000 cells were collected during a one-minute sampling time at an ultrasound intensity slightly exceeding the cavitation threshold. Furthermore, it was found that multiple cell collections can be performed within a single day using the mucosal cell collection device of the present invention.

[0042] Next, the mucosal cell collection device 201 according to the second embodiment will be described. In the first embodiment, the Langevin transducer, which is an electrostrictive ultrasonic transducer, was given as an example of the ultrasonic transducer 11a, but the embodiment is not limited to this. As shown in Figures 23 to 26, in the second embodiment, the configuration of the ultrasonic transducer 220 and the ultrasonic transmission member 222 differs from that of the first embodiment described above. In the following description, the same reference numerals are used for components similar to those in the first embodiment described above, and detailed descriptions are omitted.

[0043] Figure 23 is a perspective view of the mucosal cell collection device 201 according to the second embodiment. Figure 24 is a perspective view showing an example of applying an endoscope to the mucosal cell collection device 201 according to the second embodiment. Figure 25 is a longitudinal cross-sectional view of the mucosal cell collection device 201 according to the second embodiment. Figure 26 is a diagram illustrating the conversion of the vibration direction by the ultrasonic transmission member 222 according to the second embodiment. Referring also to Figures 23 to 26, the mucosal cell collection device 201 comprises an ultrasonic irradiation means 211, a support 230 that supports the ultrasonic irradiation means 211, a cylindrical member 213 provided around the ultrasonic irradiation means 211 and having a protrusion 212 protruding from one end of the ultrasonic irradiation means 211, an introduction means 16 for introducing liquid into the space 15 formed by the ultrasonic irradiation means 211, the protrusion 212 of the cylindrical member 213 and the subject 14, and an extraction means 17 for extracting cells separated from the mucosa 14a of the subject 14 from the space 15 together with the liquid.

[0044] The ultrasonic irradiation means 211 includes an ultrasonic transducer 220 and an ultrasonic transmission member 222 that transmits ultrasonic waves generated by the ultrasonic transducer 220. The ultrasonic transducer 220 includes a disc-shaped disc portion 221. An example of the disc portion 221 is a piezoelectric element (piezo element) that converts the force applied to a piezoelectric material into vibration. The piezoelectric element may be formed from, for example, PZT (lead zirconate titanate), a type of piezoelectric ceramic.

[0045] The ultrasonic transducer 220 is configured to vibrate laterally along the radial direction of the disc portion 221. Hereinafter, this lateral vibration will also be referred to as "lateral vibration". Electrical wiring 231 is connected to the upper and lower surfaces of the ultrasonic transducer 220. The electrical wiring 231 may be connected to the ultrasonic transducer 220 via, for example, solder 232.

[0046] For example, when a voltage is applied to the ultrasonic transducer 220 through the electrical wiring 231, the ultrasonic transducer 220 vibrates (lateral vibration) in the direction of arrow V1 in Figure 26 (lateral direction, radially inward in the example shown). The ultrasonic transmission member 222 converts the vibration of the ultrasonic transducer 220 into vibration in the vertical direction perpendicular to the lateral direction (hereinafter also referred to as "longitudinal vibration"). For example, when the ultrasonic transducer 220 vibrates in the direction of arrow V1 in Figure 26 (lateral vibration, radially inward in the example shown), the ultrasonic transmission member 222 vibrates (longitudinal vibration) in the direction of arrow V3 in Figure 26 (vertical direction, downward in the example shown).

[0047] The ultrasonic transmission member 222 is preferably made of a metal that has excellent transmission efficiency for ultrasonic waves generated by the ultrasonic transducer 220. Examples of suitable metals include those used in the first embodiment described above.

[0048] The ultrasonic transmission member 222 is formed in a cymbal shape. The ultrasonic transmission member 222 includes an outer peripheral portion 223 connected to the disc portion 221, an inclined portion 224 connected to the outer peripheral portion 223 and extending inclined away from the disc portion 221, and a convex portion 225 connected to the inclined portion 224 and convex in the opposite direction from the disc portion 221.

[0049] The outer peripheral portion 223 is formed in an annular shape along the outer circumference of the disc portion 221. The outer peripheral portion 223 may be connected to the outer circumference of the lower surface of the disc portion 221 via, for example, an adhesive 233. The inclined portion 224 is formed in a conical shape (tapered shape) that slopes downward as it moves radially inward from the inner edge of the outer circumference portion 223. For example, when the ultrasonic transducer 220 vibrates in the direction of arrow V1 in Figure 26 (lateral vibration, radially inward in the example shown), the inclined portion 224 is displaced in the direction of arrow V2 in Figure 26 (diagonal direction, radially inward and downward in the example shown). The protrusion 225 is formed in a convex shape that protrudes downward from the inner periphery of the inclined portion 224. The protrusion 225 is arranged coaxially with the disc portion 221. The outer diameter of the protrusion 225 is smaller than the outer diameter of the ultrasonic transducer 220.

[0050] The cylindrical member 213 is provided around the protrusion 225 of the ultrasonic transmission member 222. The shape of the cylindrical member 213 is not particularly limited as long as it is cylindrical, but a cylindrical shape is preferred. The cylindrical member 213 is detachably attached to the protrusion 225 of the ultrasonic transmission member 222. The cylindrical member 213 may be detachably attached to the protrusion 225, for example, via a screw structure. For example, the inner surface of the cylindrical member 213 may be provided with a female thread (not shown), and the outer surface of the protrusion 225 may be provided with a male thread (not shown). Detachability may be achieved by screwing the male thread of the protrusion 225 into the female thread of the cylindrical member 213.

[0051] It is preferable that the inner surface of the cylindrical member 213 is provided with a sealing member to prevent leakage of liquid from the space 15. An O-ring 18 is preferred as the sealing member. It is preferable that a groove 213a for mounting the O-ring 18 is provided on the inner surface of the cylindrical member 213. The sealing member may be integrally molded with the cylindrical member 213. In this case, it is preferable that a groove for mounting the sealing member is provided on the outer surface of the protrusion 225.

[0052] It is preferable that a groove 225a for mounting the O-ring 18 is provided on the outer surface of the protrusion 225. The sealing member may be integrally molded with the protrusion 225. In this case, it is preferable that a groove for mounting the sealing member is provided on the inner surface of the cylindrical member 213.

[0053] The cylindrical member 213 has through holes 213b for connecting the space 15 and the introduction means 16, and through holes 213b for connecting the space 15 and the extraction means 17. The through holes 213b and 213c open the cylindrical member 213 in the radial direction. Preferably, the through holes 213b and 213c are arranged on opposite sides of the cylindrical member 213 in the radial direction.

[0054] The cylindrical member 213 and at least a part of the introduction means 16 and / or the extraction means 17 may be formed integrally. In this case, the lumen of the introduction means 16 and the through hole 213b and / or the lumen of the extraction means 17 and the through hole 213c are integrated.

[0055] The introduction means 16 includes, for example, a tube. One end of the introduction means 16 is in communication with the space 15. The other end of the introduction means 16 may be connected to a reservoir (not shown) and a pump (not shown) for supplying liquid to be introduced into the space 15. The extraction means 17 includes, for example, a tube. One end of the extraction means 17 is in communication with the space 15. The other end of the extraction means 17 may be connected to a recovery tank (not shown) for recovering the liquid extracted from the space 15.

[0056] The support 230 is formed in a cylindrical shape. The support 230 is positioned coaxially with the ultrasonic transducer 220. The outer diameter of the support 230 may be the same as the outer diameter of the ultrasonic transducer 220. The support 230 may be connected to the upper surface of the ultrasonic transducer 220 via, for example, an adhesive 233. The support 230 supports the upper surface of the ultrasonic transducer 220 and prevents deflection deformation.

[0057] The support 230 may have, for example, the tip 240 of an endoscope (an example of an imaging device) attached to it. Preferably, the tip 240 of the endoscope is detachably attached to the side of the support 230 opposite to the ultrasonic transducer 220. In Figure 24, the outer diameters of the tip 240 of the endoscope and the support 230 are approximately equal, but this is not limited to this. For example, the outer diameters of the mucosal cell collection device 201 and the support 230 may be made smaller so as not to interfere with the illumination and observation functions of the endoscope. Alternatively, the mucosal cell collection device 201 and the support 230 may be displaced from the center and positioned above the endoscope forceps channel, etc. Furthermore, a cylindrical resin cap may be attached to the tip 240 of the endoscope. In this case, the mucosal cell collection device 201 and the support 230 may be positioned at the tip of the cap and attached to the tip 240 of the endoscope via the cap.

[0058] The size of the mucosal cell collection device 201 can be appropriately set according to the site from which mucosal cells are collected. In this embodiment, the maximum lateral dimension Wmax of the mucosal cell collection device 201 is 11 mm or less, and the maximum vertical dimension Hmax is 10 mm or less. The maximum lateral dimension Wmax of the mucosal cell collection device 201 corresponds to the diameter of the ultrasonic transducer 220. The maximum vertical dimension Hmax of the mucosal cell collection device 201 corresponds to the distance between the upper end of the support 230 and the lower end of the protrusion 212.

[0059] In the second embodiment described above, the ultrasonic transducer 220 includes a disc-shaped disc portion 221. The ultrasonic transducer 220 is configured to vibrate laterally along the radial direction of the disc portion 221. This configuration makes it easier to miniaturize the ultrasonic transducer 220 in the thickness direction (longitudinal direction) compared to when the ultrasonic transducer is a Langevin transducer (when the ultrasonic transducer includes a block body). Therefore, it contributes to miniaturization and weight reduction of the mucosal cell collection device 201 in the longitudinal direction. For example, by miniaturizing the mucosal cell collection device 201, it becomes easier to collect cells from the mucosa inside the body (e.g., the inner wall of the digestive tract such as the stomach and intestines, and the surface of abdominal organs).

[0060] In the second embodiment, the ultrasonic transmission member 222 converts the vibrations of the ultrasonic transducer 220 into vibrations in the vertical direction perpendicular to the lateral direction. With this configuration, the lateral vibration of the ultrasonic transducer 220 can be converted into longitudinal vibration, thereby generating longitudinal vibration at the tip of the device (the lower end of the protruding portion 212).

[0061] In the second embodiment, the ultrasonic transmission member 222 is formed in a cymbal shape. This configuration makes it easier to miniaturize the ultrasonic transmission member 222 in the thickness direction (vertical direction) compared to when the ultrasonic transmission member is formed in a cylindrical shape. Therefore, it contributes to miniaturization and weight reduction of the mucosal cell collection device 201 in the vertical direction.

[0062] In the second embodiment, the ultrasonic transmission member 222 includes an outer peripheral portion 223 connected to the disc portion 221, an inclined portion 224 connected to the outer peripheral portion 223 and extending inclined away from the disc portion 221, and a convex portion 225 connected to the inclined portion 224 and convex in the opposite direction from the disc portion 221. With this configuration, the lateral vibration of the disc portion 221 is converted into longitudinal vibration of the convex portion 225 by the mechanical bending deformation of the inclined portion 224 connected to the outer peripheral portion 223, thereby generating longitudinal vibration of the device tip (the lower end of the protruding portion 212).

[0063] In the second embodiment, the cylindrical member 213 is provided around the protrusion 225. This configuration allows for miniaturization of the cylindrical member 213 in conjunction with the protrusion 225, which offers significant practical benefits when applied to specific uses (for example, sampling from small animals or reducing the burden on patients).

[0064] In the second embodiment, the maximum lateral dimension Wmax of the mucosal cell collection device 201 is 11 mm or less, and the maximum vertical dimension Hmax is 10 mm or less. This configuration contributes to miniaturization and weight reduction of the mucosal cell collection device 201.

[0065] Next, the mucosal cell collection device 301 according to the third embodiment will be described. In the first embodiment, an example was given in which the ultrasonic transmission member 11b is cylindrical and the tubular member 13 is cylindrical, but the embodiment is not limited to this. As shown in Figures 27 to 29, in the third embodiment, the configuration of the ultrasonic transmission member 322 and the tubular member 313 differs from that of the first embodiment described above. In the following description, the same reference numerals are used for components similar to those in the first embodiment described above, and detailed descriptions are omitted.

[0066] Figure 27 is a longitudinal cross-sectional view of the mucosal cell collection device 301 according to the third embodiment. Figure 28 is an end view of the mucosal cell collection device 301 of Figure 27, viewed from the lower end. Figure 29 is a diagram illustrating the mounting position of the cylindrical member 313 and / or sealing member according to the third embodiment. In Figures 27 and 29, the cylindrical member 313 of the mucosal cell collection device 301 is shown with a cross-sectional hatch. Referring also to Figures 27 to 29, the mucosal cell collection device 301 comprises an ultrasonic irradiation means 311, a cylindrical member 313 provided around the ultrasonic irradiation means 311 and having a protrusion 312 protruding from one end of the ultrasonic irradiation means 311, an introduction means 16 for introducing liquid into the space 15 formed by the ultrasonic irradiation means 311, the protrusion 312 of the cylindrical member 313 and the subject 14, and an extraction means 17 for extracting cells separated from the mucosa 14a of the subject 14 together with the liquid from the space 15.

[0067] The ultrasonic irradiation means 311 comprises an ultrasonic transducer 11a and an ultrasonic transmission member 322 that transmits ultrasonic waves generated by the ultrasonic transducer 11a. The ultrasonic transmission member 322 is formed in such a shape that the cross-sectional area perpendicular to the axial direction of the ultrasonic transmission member 322 decreases towards the tip of the protrusion 312. The cross-sectional area perpendicular to the axial direction of the ultrasonic transmission member 322 corresponds to the area of ​​the ultrasonic transmission member 322 when viewed in cross-section (cross-sectional area).

[0068] The ultrasonic transmission member 322 in this embodiment corresponds to a horn. A horn is a type of transmission body in which the cross-sectional area in the longitudinal direction changes, and it is responsible not only for transmitting vibrations but also for modifying the vibration amplitude.

[0069] The ultrasonic transmission member 322 is formed in an exponential shape, for example, with straight sections connected to a large end face and a small end face. In this embodiment, the ultrasonic transmission member 322 includes a first straight section 323 formed in the shape of a cylindrical shape extending in a straight line, a constricted section 324 connected to the lower end of the first straight section 323 and decreasing in diameter as it extends downwards, and a second straight section 325 formed in the shape of a cylindrical shape extending in a straight line connected to the lower end of the constricted section 324. Figure 27 shows, as an example of the dimensions of the ultrasonic transmission member 322, the vertical length of the first straight section 323, the vertical length of the constricted section 324, and the vertical length of the second straight section 325 (the upper reference line of dimension 20.87 shown in Figure 27 is the starting point of the straight section of the horn and is approximately the position where the extraction means 17 is bent) (units [mm] are not shown), but the lengths of each part are not limited to those shown above.

[0070] The form of the ultrasonic transmission member 322 is not limited to those described above. For example, the ultrasonic transmission member 322 may be formed in the form of a step, conical, simple exponential, catenoidal, Fourier, stepped composite, or the like.

[0071] When a Langevin transducer is provided as the ultrasonic transducer 11a, it is preferable that the ultrasonic transmission member 322 has a shape that resonates at the resonant frequency of the Langevin transducer. It is preferable that the ultrasonic transmission member 322 has a shape that is easy to insert into the oral cavity. It is preferable that the ultrasonic transmission member 322 is made of a material (for example, a titanium alloy) that has excellent fatigue strength, vibration characteristics, and corrosion resistance.

[0072] The cylindrical member 313 is formed in such a shape that the cross-sectional area perpendicular to the axial direction of the cylindrical member 313 decreases towards the tip of the protruding portion 312. The cross-sectional area perpendicular to the axial direction of the cylindrical member 313 corresponds to the area of ​​the cylindrical member 313 when viewed in cross-section (cross-sectional area).

[0073] The cylindrical member 313 is formed in a shape that follows, for example, a part of the outer shape of the ultrasonic transmission member 322. In this embodiment, the cylindrical member 313 includes a reduced-diameter portion 314 that follows the lower part of the constricted portion 324 of the ultrasonic transmission member 322 and decreases in diameter as it goes downwards, and a straight tube portion 315 that follows the second straight portion 325 of the ultrasonic transmission member 322, connects to the lower end of the reduced-diameter portion 314, and is formed in a cylindrical shape that extends in a straight line. However, the form of the cylindrical member 313 is not limited to the above. In Figure 27, as an example of the dimensions of the cylindrical member 313, the vertical length of the portion of the cylindrical member 313 excluding the protruding portion 312 is shown (unit [mm] is not shown), but it is not limited to the above. Figure 28 shows, as an example of the dimensions of the cylindrical member 313, the length in the long axis and uniaxial direction of the cylindrical member 313 as viewed from the lower end of the mucosal cell collection device 301, and the inner diameter of the straight tube section 315 (corresponding to the diameter d1 of the approximately cylindrical space 15 described above) (units [mm] are not shown), but the dimensions are not limited to those shown.

[0074] The cylindrical member 313 is attached to the ultrasonic transmission member 322 at a position NP that corresponds to a node when the ultrasonic transmission member 322 vibrates due to the ultrasonic waves generated by the ultrasonic transducer 11a. The position NP that corresponds to a node when the ultrasonic transmission member 322 vibrates corresponds to a position where the ultrasonic transmission member 322 does not vibrate when a standing wave is applied to it (a position different from the position that corresponds to an antinode where it vibrates).

[0075] The mucosal cell collection device 301 includes an O-ring 18 (an example of a sealing member) between the ultrasonic transmission member 322 and the cylindrical member 313. The O-ring 18 is attached to the ultrasonic transmission member 322 and / or the cylindrical member 313 at position NP corresponding to the aforementioned section.

[0076] In this embodiment, a groove 313a for mounting an O-ring 18 is provided on the inner surface of the cylindrical member 313. The groove 313a is provided on the upper inner circumferential surface of the reduced diameter portion 314 of the cylindrical member 313. However, the form of the groove 313a is not limited to that described above.

[0077] In this embodiment, the O-ring 18 is attached to the constricted portion 324 of the ultrasonic transmission member 322 and the reduced diameter portion 314 of the cylindrical member 313 at position NP corresponding to the aforementioned section. However, the mounting configuration of the O-ring 18 is not limited to the above.

[0078] In the third embodiment described above, the ultrasonic transmission member 322 is formed in such a shape that the cross-sectional area perpendicular to the axial direction of the ultrasonic transmission member 322 decreases towards the tip of the protrusion 312. This configuration offers significant advantages when the tip of the sampling device can be made thinner to suit specific applications (for example, sampling from small animals or reducing the burden on patients).

[0079] In the third embodiment, the cylindrical member 313 is formed in such a shape that the cross-sectional area perpendicular to the axial direction of the cylindrical member 313 decreases towards the tip of the protruding portion 312. This configuration offers significant practical benefits when the tip of the sampling device is made thinner to suit specific applications (for example, sampling from small animals or reducing the burden on patients).

[0080] In the third embodiment, the cylindrical member 313 is attached to the ultrasonic transmission member 322 at a position NP that corresponds to a node when the ultrasonic transmission member 322 vibrates due to the ultrasonic waves generated by the ultrasonic transducer 11a. This configuration allows for efficient transmission of vibrations from the ultrasonic transmission member 322 to the cylindrical member 313.

[0081] In the third embodiment, an O-ring 18 is provided between the ultrasonic transmission member 322 and the cylindrical member 313, and the O-ring 18 is attached to the ultrasonic transmission member 322 and the cylindrical member 313 at position NP corresponding to the aforementioned section. With this configuration, even when the O-ring 18 is provided, the vibrations of the ultrasonic transmission member 322 can be efficiently transmitted to the cylindrical member 313.

[0082] The present invention will be described in more detail below with an example of the mounting position of the sealing member, but the present invention is not limited to the example of the mounting position of the sealing member described later.

[0083] Figure 30 shows the mounting positions of the sealing member, from position (1) to position (4). Figure 31 shows the relationship between the mounting position of the sealing member and the relative value of the output ultrasonic intensity. As shown in Figure 30, position (4) corresponds to the position of the aforementioned section. The positions approach the position of the aforementioned section in the order from position (1) to position (4).

[0084] The vertical axis in Figure 31 shows the relative value with the intensity at position (1) set to 1. As shown in Figure 31, it was found that the intensity increases as the sample approaches the position corresponding to the aforementioned section (position (4)). [Explanation of symbols]

[0085] 1,201,301 Mucosal cell collection device 11,211,311 Ultrasonic irradiation means 11a,220 Ultrasonic transducer 11b,222,322 Ultrasonic transmission member 12,212,312 Projection 13,213,313 Cylindrical member 13a,213a,313a Groove 13b,13c,213b,213c through hole 14 Subjects 14a mucosa 14b cells 15 Space 16. Introduction Methods 17 Retrieval means 18 O-rings 221 Disc section 223 Outer perimeter 224 Slope 225 Convex part Hmax: Maximum vertical dimension The position corresponding to the NP node W-Float Maximum horizontal dimension

Claims

1. Ultrasonic irradiation means, A cylindrical member provided around the ultrasonic irradiation means and having a protruding portion that extends from one end of the ultrasonic irradiation means, An introduction means for introducing liquid into the space formed by the ultrasonic irradiation means, the protruding portion of the cylindrical member, and the subject, and Extraction means for removing cells separated from the mucous membrane of the subject along with the liquid from the space. A mucosal cell collection device equipped with the following features.

2. The mucosal cell collection apparatus according to claim 1, wherein the ultrasonic irradiation means comprises an ultrasonic transducer and an ultrasonic transmission member that transmits ultrasonic waves generated by the ultrasonic transducer.

3. The mucosal cell collection apparatus according to claim 1 or 2, wherein the introduction means and the extraction means each comprise a tube.

4. The mucosal cell collection apparatus according to claim 1 or 2, further comprising a sealing member between the ultrasonic irradiation means and the cylindrical member.

5. The mucosal cell collection device according to claim 2, wherein the ultrasonic transducer includes a disc-shaped portion and is configured to vibrate laterally along the radial direction of the disc portion.

6. The mucosal cell collection apparatus according to claim 5, wherein the ultrasonic transmission member converts the vibration of the ultrasonic transducer into vibration in a vertical direction perpendicular to the lateral direction.

7. The mucosal cell collection device according to claim 6, wherein the ultrasonic transmission member is formed in the shape of a cymbal.

8. The mucosal cell collection device according to claim 7, wherein the ultrasonic transmission member comprises an outer peripheral portion connected to the disc portion, an inclined portion connected to the outer peripheral portion and extending inclined away from the disc portion, and a convex portion connected to the inclined portion and convex in the opposite direction from the disc portion.

9. The mucosal cell collection device according to claim 8, wherein the cylindrical member is provided around the protrusion.

10. The maximum horizontal dimension is 11 mm or less. The mucosal cell collection device according to claim 9, wherein the maximum vertical dimension is 10 mm or less.

11. The mucosal cell collection device according to claim 2, wherein the ultrasonic transmission member is formed in a shape in which the cross-sectional area perpendicular to the axial direction of the ultrasonic transmission member decreases towards the tip of the protrusion.

12. The mucosal cell collection device according to claim 11, wherein the cylindrical member is formed in such a shape that the cross-sectional area perpendicular to the axial direction of the cylindrical member decreases towards the tip of the protruding portion.

13. The mucosal cell collection device according to claim 2, wherein the cylindrical member is attached to the ultrasonic transmission member at a position corresponding to a node when the ultrasonic transmission member vibrates due to the ultrasonic waves generated by the ultrasonic transducer.

14. A sealing member is provided between the ultrasonic transmission member and the cylindrical member. The mucosal cell collection device according to claim 13, wherein the sealing member is attached to the ultrasonic transmission member and / or the cylindrical member at the position corresponding to the node.

15. Ultrasonic irradiation means, A cylindrical member provided around the ultrasonic irradiation means and having a protruding portion that extends from one end of the ultrasonic irradiation means, An introduction means for introducing liquid into the space formed by the ultrasonic irradiation means, the protruding portion of the cylindrical member, and the subject, and Extraction means for removing cells separated from the mucous membrane of the subject along with the liquid from the space. A method for collecting mucosal cells using a mucosal cell collection device, comprising: The steps of irradiating the mucous membrane of the subject with ultrasound through the liquid, and The process of removing cells separated from the mucous membrane of the subject together with the liquid. A method for collecting mucosal cells, comprising the following features.

16. Furthermore, the method for collecting mucosal cells according to claim 15, comprising the step of filling the space formed by the ultrasonic irradiation means, the protruding portion of the cylindrical member, and the subject with the liquid and circulating it, before or at the beginning of the irradiation step, and washing and removing oral substance in the space and / or oral substance adhering to the mucosal surface of the subject with the liquid.

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