Cell examination method using gelation of alcohol solution composition

Abstract

The present invention relates to a technology for gelling a sample (e.g., exfoliated human cells) in a vial, safely transporting the sample to a testing center, restoring the sample to its original state (e.g., converting it to a liquid phase) at the testing center, and then examining the sample. More specifically, the present invention relates to a technology for converting a sample-preserving liquid contained in a vial into a jelly state, safely transporting the sample to a testing center without deforming it, redissolving the sample-preserving liquid into a liquid phase at the testing center, and then examining the sample. The present invention is advantageous in that gelling of the cell-fixing liquid filled in the vial is simplified because the cell-fixing liquid is gelled by mixing a selected solution with a selected material.

Description

Technical Field

[0001] The present invention relates to a technology for gelling a sample (e.g., exfoliated cells from a human body) in a vial, safely transporting the sample to an inspection center, restoring the sample to its original state (e.g., converting it into a liquid phase) at the inspection center, and inspecting the sample.

[0002] More specifically, the present invention relates to a technology for converting a sample preservation liquid contained in a vial into a jelly state, safely transporting the sample to an inspection center without deformation, redissolving the sample preservation liquid into a liquid phase at the inspection center, and inspecting the sample. Background Art

[0003] Generally, a sample (exfoliated cells) is obtained from a human body and then spread on a slide for cytodiagnosis at a testing center. The state of the exfoliated cells is then observed for diagnosis.

[0004] Patients who have difficulty visiting a testing center should collect their own exfoliated cells (samples) and then transport them to the testing center in an intact state. To do this, the sample can be placed in a sealed vial along with a cell fixation liquid to protect it and then transported to the testing center.

[0005] The cell fixation liquid filled in the vial to prevent sample deformation is generally composed of an aqueous solution. During the transportation of the vial, the cell fixation liquid often leaks through the gap between the vial body and the vial cap, which can cause sample deformation.

[0006] There is a need to develop a technology that can solve these problems of the existing technology.

[0007] Meanwhile, existing documents related to the present invention are as follows.

[0008] (1) EP 0511430 A2 (November 4, 1992) "Cell preservative solution"

[0009] (2)US 2006 / 0088814 A1 (April 27, 2006) "Enhanced cell preservativesolution and methods for using same"

[0010] (3) Korean Patent Application Publication No. 10-2005-0116689 (December 13, 2005) "Gel preservative, method for preparing the same, and cell examining method using the same" Summary of the Invention

[0011] Technical issues

[0012] The present invention is proposed in view of the above-mentioned viewpoints, and its purpose is to provide a cell examination method using gelation of an alcohol solution composition. Through this method, the sample contained in a vial (for example, exfoliated cells from the human body) is gelled with the alcohol solution composition so that the sample can be safely transported over long distances and can be easily restored to its original state when the cell examination is performed at the examination center.

[0013] Technical Solution

[0014] To achieve the above object, the cell examination method using gelation of an alcohol solution composition according to the first embodiment of the present invention may be configured to include: (a) mixing any one of a first solution containing 30 to 60 parts by weight of a methanol aqueous solution, 0.1 to 0.2 parts by weight of EDTA, 0.01 to 0.05 parts by weight of bile acid, and 0.05 to 0.1 parts by weight of a 1N acetic acid aqueous solution or a second solution containing 40 to 50 parts by weight of an ethanol aqueous solution, 0.1 to 0.2 parts by weight of EDTA, and 0.2 to 1 parts by weight of a glycol additive with 5 to 20 parts by weight of one or more of gelatin or a photocurable protein; (a) mixing and gelling any one of a natural cellulose polymer material, 1 to 100 parts by weight of a synthetic acrylic polymer material bead at a temperature of 1° C. to 40° C. to produce an alcoholic jelly-like material; (b) placing a collected sample into a sealed container containing the alcoholic jelly-like material; (d) adding the gelled alcoholic jelly-like material in the sealed container to a re-dissolution solution; (e) maintaining the re-dissolution solution at a temperature of 1° C. to 40° C. to re-dissolve the alcoholic jelly-like material into a liquid phase; and (f) obtaining a sample from the re-dissolved alcoholic jelly-like material and examining the sample.

[0015] The cell examination method using gelation of an alcohol solution composition according to the second embodiment of the present invention may be configured to include: (a) placing any one of a first solution containing 30 to 60 parts by weight of a methanol aqueous solution, 0.1 to 0.2 parts by weight of EDTA, 0.01 to 0.05 parts by weight of bile acid, and 0.05 to 0.1 parts by weight of a 1N acetic acid aqueous solution or a second solution containing 40 to 50 parts by weight of an ethanol aqueous solution, 0.1 to 0.2 parts by weight of EDTA, and 0.2 to 1 parts by weight of a glycol additive (hereinafter referred to as the "selected solution") in a sealed container; (b) placing a sample in the sealed container filled with the selected solution; and (c) heating the sample at 1°C to 40°C. (d) adding the gelled alcoholic jelly-like material in the sealed container to a re-dissolving solution; (e) maintaining the re-dissolving solution at a temperature of 1°C to 40°C to re-dissolve the gelled alcoholic jelly-like material into a liquid material; and (f) obtaining a sample from the re-dissolved liquid material and examining the sample.

[0016] At this time, step (d) may include the step of preparing a re-dissolution solution comprising 30 to 60 parts by weight of a methanol aqueous solution, 0.1 to 0.2 parts by weight of EDTA, 0.01 to 0.05 parts by weight of bile acid, and 0.05 to 0.1 parts by weight of a 1N acetic acid aqueous solution.

[0017] In addition, step (d) may include the step of preparing a solution for redissolution, the solution being configured to include 40 to 50 parts by weight of an ethanol aqueous solution, 0.1 to 0.2 parts by weight of EDTA, and 0.2 to 1 parts by weight of a glycol additive.

[0018] Beneficial effects

[0019] The present invention has an advantage in that gelation of the cell-fixing liquid filled in the interior of the vial is simple because the cell-fixing liquid is gelled by mixing a selected solution and a selected material.

[0020] The present invention has the advantage that, by gelling the cell fixation liquid, leakage of the cell fixation liquid from the vial during long-distance transportation can be prevented.

[0021] The present invention is also advantageous in that the conversion from the gelled state to the liquid phase for cell examination is simple. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 is an illustrative view of a vial, which is a closed container that contains a sample for transport.

[0023] Figure 2 It is an exemplary view of a state where a vial is mounted on a stirrer for gelation.

[0024] Figure 3 It is an exemplary view of a state where a vial is mounted on a stirrer for reconstitution.

[0025] Figure 4 This is a flowchart showing a cell examination process using gelation of an alcohol solution composition according to the first embodiment of the present invention.

[0026] Figure 5 This is a flowchart showing a cell examination process using gelation of an alcohol solution composition according to a second embodiment of the present invention. DETAILED DESCRIPTION

[0027] Hereinafter, the present invention will be described in detail.

[0028] Figure 1 is an illustrative view of a vial, which is a sealed container that contains a sample for transport. Figure 1 The vial 10 may include a hollow body 12 having one end opened and a cap 11 that opens and closes the opening portion of the body 12 .

[0029] In this case, the vial 10 can be made by extruding a resin containing an antioxidant, because the alcohol component may be oxidized when the vial 10 containing the cell fixation liquid is exposed to light or oxygen. The vial 10 can also be made translucent by adding a light-blocking dye.

[0030] By using an opaque bag-type packaging material as a container for the liquid for cell fixation, exposure to oxygen and light can be prevented.

[0031] At the same time, the portion of the sample not immersed in the cell fixation liquid may be damaged during the process of placing a brush tip (not shown) for collecting the sample into the vial 10. Therefore, it is preferable to manufacture the vial 10 to a size that can completely accommodate the brush tip. It is also preferable to completely fill the vial 10 with the cell fixation liquid while the brush tip for collecting the sample is placed inside the vial 10.

[0032] Conventionally, when a cell-fixing liquid in the form of an aqueous solution is filled into a vial 10 along with a sample, leakage during long-distance transport can cause sample deformation. To address this issue, the present invention provides a composition capable of gelling the cell-fixing liquid. In other words, when the cell-fixing liquid is gelled, the likelihood of leakage or evaporation is low, and the original state of the sample can be maintained.

[0033] Here, the cell fixation liquid contained in the vial 10 is transported to the examination center in a jelly state, and the gelled state is maintained by keeping the vial 10 at a low temperature of room temperature or lower during transportation.

[0034] Figure 2 1 is an exemplary view of a state where the vial 10 is mounted on the stirrer 20 for gelation. Figure 2 , a stirrer 20 for gelation may be provided as a means for mixing the selected solution and the selected material of the present invention to produce an alcohol-based jelly-like material.

[0035] First, the vial 10 is mounted on the table member 21 of the blender 20 for gelling. In this state, the blender 20 for gelling is operated to position the table member 21 below the injection member 22, and the jelly composition is filled into the vial 10 as it drips downward from the injection member 22.

[0036] The jelly composition in the vial 10 can be gelled by operating the stirrer 20 and thereby moving the workbench member 21 along the Figure 2 Mix thoroughly by moving back and forth in the direction of the arrow.

[0037] Figure 3 This is an exemplary view showing a state where the vial 10 is mounted on the stirrer 30 for redissolution. When the alcoholic jelly material of the present invention arrives at the testing center in a state contained together with a sample in the vial 10, it is necessary to convert the alcoholic jelly material in the vial 10 into a liquid phase before removing the sample from the vial 10.

[0038] To this end, the vial 10 is mounted on the table member 31 of the stirrer 30 for reconstitution. In this state, the stirrer 30 for reconstitution is operated to position the table member 31 below the injection member 32, and as the reconstitution solution drips downward from the injection member 32, the alcohol jelly-like material in the vial 10 is converted back into a liquid phase.

[0039] At this time, by operating the stirrer 30 for redissolving and thereby moving the table member 31 along the Figure 3 The reconstitution solution in the vial 10 and the jelly composition can be thoroughly mixed by reciprocating the movement in the direction of the arrow in the vial.

[0040] In this manual, for the sake of convenience, Figure 2 Agitator 20 for gelation and Figure 3 The stirrer 30 for re-dissolving is described with different names, but the same equipment can be used. Figure 2 Agitator 20 for gelation and Figure 3 The stirrer 30 for re-dissolution is shown in a similar form, but may be configured in a different form.

[0041] Figure 4 This is a flowchart showing a cell examination process using gelation of an alcohol solution composition according to the first embodiment of the present invention.

[0042] Step S110: First, select any one solution (hereinafter referred to as "selected solution") from a first solution containing 30 to 60 parts by weight of a methanol aqueous solution, 0.1 to 0.2 parts by weight of EDTA (ethylenediaminetetraacetic acid), 0.01 to 0.05 parts by weight of bile acid, and 0.05 to 0.1 parts by weight of a 1N acetic acid aqueous solution, or a second solution containing 40 to 50 parts by weight of an ethanol aqueous solution, 0.1 to 0.2 parts by weight of EDTA, and 0.2 to 1 parts by weight of a glycol additive.

[0043] In addition, any one material is selected from 5 to 20 parts by weight of one or more of gelatin or photocurable protein (methacrylated gelatin; GelMA), 1 to 100 parts by weight of natural cellulose polymer material, or 1 to 100 parts by weight of synthetic acrylic polymer material beads (hereinafter referred to as "selected material").

[0044] Subsequently, the selected solution and the selected material are placed in the vial 10 and then mixed at a temperature of 1° C. to 40° C. for gelation to produce an alcohol-based jelly-like material.

[0045] At this time, when the temperature around the vial 10 is as low as less than 1° C., gelatin, photocurable protein, natural cellulose polymer material, and synthetic acrylic polymer beads are not dissolved in the vial 10 , resulting in poor gelation or jellification.

[0046] When the temperature around the vial 10 is high exceeding 40° C., the alcohol component having a low boiling point evaporates, thereby causing the physical properties of the entire composition to change and the sample fixing ability of the cell fixing means to become excessively strong or weak.

[0047] Step S120: Next, the sample is placed in the vial 10 containing the alcohol jelly material. At this time, the vial 10 is preferably made into a size that can fully accommodate a brush head (not shown) for collecting the sample, so that the brush head can be safely placed in the vial 10.

[0048] In this way, the vial 10 containing the sample is transported to the inspection center. During transportation, the vial 10 needs to be kept at a low temperature of room temperature or below to prevent the jelly state from being converted into a liquid phase.

[0049] When the vial 10 containing the sample is transported, the sample needs to be removed from the vial 10 and subjected to cell testing at a testing center. In order to safely remove the sample from the vial 10 without deformation, the cell fixation means in the vial 10 needs to be converted back into a liquid phase.

[0050] Steps S130 and S140: The gelled alcoholic jelly-like material in the vial 10 may be redissolved into a liquid phase by adding the alcoholic jelly-like material to a redissolution solution and then maintaining the redissolution solution at a temperature of 1°C to 40°C.

[0051] At this time, the vial 10 containing the jelly-like material and the sample is placed in a container. Figure 3 It is possible to implement an embodiment in which the reconstitution solution is dripped into the interior of the vial 10 while being mounted on the mechanism of the stirrer 30 for reconstitution. It is also possible to implement an embodiment in which the vial 10 is placed in the reconstitution solution separately from the stirrer 30 for reconstitution.

[0052] Meanwhile, the re-dissolution solution in the first embodiment of the present invention may be configured to include 30 to 60 parts by weight of a methanol aqueous solution, 0.1 to 0.2 parts by weight of EDTA, 0.01 to 0.05 parts by weight of bile acid, and 0.05 to 0.1 parts by weight of a 1N acetic acid aqueous solution.

[0053] The re-dissolution solution in the first embodiment of the present invention may also be configured to include 40 to 50 parts by weight of an ethanol aqueous solution, 0.1 to 0.2 parts by weight of EDTA, and 0.2 to 1 parts by weight of a glycol additive.

[0054] Step S150: Figure 3 As shown, after the material in the vial 10 is converted back into a liquid phase, a sample in the vial 10 can be removed using a separate device (not shown) and subjected to cytological examination.

[0055] In a first embodiment of the present invention, the selected solution is selected from a first solution comprising 30 to 60 parts by weight of a methanol aqueous solution, 0.1 to 0.2 parts by weight of EDTA, 0.01 to 0.05 parts by weight of bile acid, and 0.05 to 0.1 parts by weight of a 1N acetic acid aqueous solution, or a second solution comprising 40 to 50 parts by weight of an ethanol aqueous solution, 0.1 to 0.2 parts by weight of EDTA, and 0.2 to 1 parts by weight of a glycol additive. The selected material is selected from one or more of 5 to 20 parts by weight of gelatin or a photocurable protein, 1 to 100 parts by weight of a natural cellulose polymer material, or 1 to 100 parts by weight of synthetic acrylic polymer material beads.

[0056] With the first embodiment of the present invention as described above, physical properties depending on the constituent ratio were studied, and while changing the mixing ratio of the individual components, the influence of the individual components on the physical properties of the entire composition was studied.

[0057] Table 1 shows experimental conditions in which only the mixing ratio of the methanol aqueous solution was changed and the mixing ratios of the other components were fixed in the first embodiment of the present invention.

[0058] [Table 1]

[0059]

[0060] Table 2 shows experimental conditions in which only the mixing ratio of EDTA was changed and the mixing ratios of other components were fixed in the first embodiment of the present invention.

[0061] [Table 2]

[0062]

[0063] Table 3 shows the experimental conditions in which only the mixing ratio of bile acid was changed and the mixing ratios of other components were fixed in the first embodiment of the present invention.

[0064] [Table 3]

[0065]

[0066] Table 4 shows experimental conditions in which only the mixing ratio of the 1N acetic acid aqueous solution was changed and the mixing ratios of the other components were fixed in the first embodiment of the present invention.

[0067] [Table 4]

[0068]

[0069] Table 5 shows experimental conditions in which only the mixing ratio of gelatin was changed and the mixing ratios of other components were fixed in the first embodiment of the present invention.

[0070] [Table 5]

[0071]

[0072] In Tables 1 to 5, the gel-based cell-fixing means exhibited excellent fixation capabilities in the Examples (A1 to A5, B1 to B5, and C1 to C5). On the other hand, the Comparative Examples (a1 to a5 and b1 to b5) exhibited either excessive or weak fixation capabilities for the sample within the vial 10. Consequently, these Comparative Examples experienced problems (e.g., sample deformation) during sample removal (e.g., re-dissolution) from the vial 10.

[0073] Regarding the first embodiment of the present invention, experiments other than those shown in Tables 1 to 5 were conducted, but descriptions of the other experiments are omitted for convenience of explanation.

[0074] Meanwhile, in the alcohol-based jelly-like material of the first embodiment of the present invention, 5 to 20 parts by weight of any one or more of gelatin or photocurable protein may be replaced with 1 to 100 parts by weight of a natural cellulose polymer (e.g., hydroxypropyl methylcellulose (mecellose) or hydroxyethyl cellulose (hecellose)) or 1 to 100 parts by weight of a synthetic acrylic polymer bead (e.g., methyl methacrylate-co-ethylene glycol dimethacrylate; MMA).

[0075] Here, also for the natural cellulose polymer, experiments were conducted with the mixing ratio of the natural cellulose polymer ranging from less than 1 part by weight to greater than 100 parts by weight as a comparative example, while the mixing ratios of other components were fixed. The result was that in the range outside 1 to 100 parts by weight, the ability to fix the sample inside the vial 10 was poor.

[0076] In addition, for the synthetic acrylic polymer beads, experiments were conducted as comparative examples with mixing ratios of the synthetic acrylic polymer beads ranging from less than 1 part by weight to greater than 100 parts by weight, while the mixing ratios of the other components were fixed. As a result, the ability to fix the sample inside the vial 10 was poor in the range outside the range of 1 to 100 parts by weight.

[0077] Meanwhile, the redissolution solution used in step S130 in the first embodiment of the present invention may be configured to include 30 to 60 parts by weight of a methanol aqueous solution, 0.1 to 0.2 parts by weight of EDTA, 0.01 to 0.05 parts by weight of bile acid, and 0.05 to 0.1 parts by weight of a 1N acetic acid aqueous solution (hereinafter referred to as the "first redissolution solution").

[0078] In the first embodiment of the present invention, physical properties depending on the constituent ratio of the first re-dissolution solution were studied, and the influence of the individual components on the physical properties of the entire composition was studied while changing the mixing ratio of the individual components.

[0079] Table 6 shows experimental conditions in which only the mixing ratio of the methanol aqueous solution was changed and the mixing ratios of the other components were fixed during the redissolution process using the first redissolution solution in the first embodiment of the present invention.

[0080] [Table 6]

[0081]

[0082] Table 7 shows the experimental conditions in which only the mixing ratio of EDTA was changed and the mixing ratios of other components were fixed during the reconstitution process using the first reconstitution solution in the first embodiment of the present invention.

[0083] [Table 7]

[0084]

[0085]

[0086] Table 8 shows experimental conditions in which only the mixing ratio of bile acid was changed and the mixing ratios of other components were fixed during the reconstitution process using the first reconstitution solution in the first embodiment of the present invention.

[0087] [Table 8]

[0088]

[0089] Table 9 shows experimental conditions in which only the mixing ratio of the 1N acetic acid aqueous solution was changed and the mixing ratios of the other components were fixed during the re-dissolution process using the first re-dissolution solution in the first embodiment of the present invention.

[0090] [Table 9]

[0091]

[0092] In Tables 6 to 9, in Examples (A6 to A9, B6 to B9, and C6 to C9), there were no particular problems in the process of removing the sample from the vial 10 (e.g., re-dissolution). On the other hand, in Comparative Examples (a6 to a9 and b6 to b9), problems (e.g., deformation of the sample) occurred in the process of removing the sample from the vial 10 (e.g., re-dissolution).

[0093] Meanwhile, the redissolution solution in the first embodiment of the present invention may be configured to include 40 to 50 parts by weight of an ethanol aqueous solution, 0.1 to 0.2 parts by weight of EDTA, and 0.2 to 1 parts by weight of a glycol additive (hereinafter referred to as "second redissolution solution").

[0094] In the first embodiment of the present invention, physical properties depending on the constituent ratio of the second redissolution solution were studied, and the influence of the individual components on the physical properties of the entire composition was studied while changing the mixing ratio of the individual components.

[0095] Table 10 shows experimental conditions in which only the mixing ratio of the ethanol aqueous solution was changed and the mixing ratios of the other components were fixed during the re-dissolution process using the second re-dissolution solution in the first embodiment of the present invention.

[0096] [Table 10]

[0097]

[0098]

[0099] Table 11 shows the experimental conditions in which only the mixing ratio of EDTA was changed and the mixing ratios of other components were fixed during the reconstitution process using the second reconstitution solution in the first embodiment of the present invention.

[0100] [Table 11]

[0101]

[0102] Table 12 shows experimental conditions in which only the mixing ratio of the glycol additive was changed and the mixing ratios of the other components were fixed during the redissolution process using the second redissolution solution in the first embodiment of the present invention.

[0103] [Table 12]

[0104]

[0105] In Tables 10 to 12, in Examples (A10 to A12, B10 to B12, and C10 to C12), no particular problems occurred during the process of removing the sample from the vial 10 (e.g., re-dissolution). On the other hand, in Comparative Examples (a10 to a12 and b10 to b12), problems (e.g., deformation of the sample) occurred during the process of removing the sample from the vial 10 (e.g., re-dissolution).

[0106] Figure 5 This is a flowchart showing a cell examination process using gelation of an alcohol solution composition according to a second embodiment of the present invention.

[0107] Step S210: First, select any one solution (hereinafter referred to as "selected solution") from a first solution containing 30 to 60 parts by weight of a methanol aqueous solution, 0.1 to 0.2 parts by weight of EDTA, 0.01 to 0.05 parts by weight of bile acid, and 0.05 to 0.1 parts by weight of a 1N acetic acid aqueous solution, or a second solution containing 40 to 50 parts by weight of an ethanol aqueous solution, 0.1 to 0.2 parts by weight of EDTA, and 0.2 to 1 parts by weight of a glycol additive, and then place the selected solution into the vial 10.

[0108] Step S220: Next, the sample is placed in the vial 10. At this time, the vial 10 is preferably manufactured to a size that can completely accommodate the brush head.

[0109] Step S230: Subsequently, any one material (hereinafter referred to as "selected material") is selected from 5 to 20 parts by weight of one or more of gelatin or photocurable protein, 1 to 100 parts by weight of natural cellulose polymer material, or 1 to 100 parts by weight of synthetic acrylic polymer material beads.

[0110] Subsequently, the selected material is put into the interior of the vial 10 in a state of containing the sample while the temperature around the vial 10 is maintained at 1° C. to 40° C., and the mixture is gelled into an alcohol-based jelly-like material.

[0111] At this time, when the temperature around the vial 10 is as low as less than 1° C., gelatin, photocurable protein, natural cellulose polymer material, and synthetic acrylic polymer are not dissolved in the vial 10 , resulting in gelation or poor gelation.

[0112] When the temperature around the vial 10 is high exceeding 40° C., the alcohol component having a low boiling point evaporates, thereby causing the physical properties of the entire composition to change and the sample fixing ability of the cell fixing means to become excessively strong or weak.

[0113] After this, the vial 10 containing the sample is transported to an inspection center. During transportation, the vial 10 needs to be kept at a low temperature of room temperature or below to prevent the jelly state from being converted into a liquid phase.

[0114] When the vial 10 containing the sample is transported, the sample needs to be removed from the vial 10 and subjected to cell testing at a testing center. In order to safely remove the sample from the vial 10 without deformation, the cell fixation means in the vial 10 needs to be converted back into a liquid phase.

[0115] Steps S240 and S250: The gelled alcoholic jelly-like material in the vial 10 may be redissolved into a liquid phase by adding the alcoholic jelly-like material to a redissolution solution and then maintaining the redissolution solution at a temperature of 1°C to 40°C.

[0116] At this time, the vial 10 containing the jelly-like material and the sample is placed in a container. Figure 3 It is possible to implement an embodiment in which the reconstitution solution is dripped into the interior of the vial 10 while being mounted on the mechanism of the stirrer 30 for reconstitution. It is also possible to implement an embodiment in which the vial 10 is placed in the reconstitution solution separately from the stirrer 30 for reconstitution.

[0117] Meanwhile, the re-dissolution solution in the second embodiment of the present invention may be configured to include 30 to 60 parts by weight of a methanol aqueous solution, 0.1 to 0.2 parts by weight of EDTA, 0.01 to 0.05 parts by weight of bile acid, and 0.05 to 0.1 parts by weight of a 1N acetic acid aqueous solution.

[0118] The re-dissolution solution in the second embodiment of the present invention may also be configured to include 40 to 50 parts by weight of an ethanol aqueous solution, 0.1 to 0.2 parts by weight of EDTA, and 0.2 to 1 parts by weight of a glycol additive.

[0119] Step S260: Figure 3 As shown, after the material in the vial 10 is converted back into a liquid phase, a sample in the vial 10 can be removed using a separate device (not shown) and subjected to cytological examination.

[0120] In a second embodiment of the present invention, the selected solution is selected from a first solution comprising 30 to 60 parts by weight of an aqueous methanol solution, 0.1 to 0.2 parts by weight of EDTA, 0.01 to 0.05 parts by weight of bile acid, and 0.05 to 0.1 parts by weight of a 1N aqueous acetic acid solution, or a second solution comprising 40 to 50 parts by weight of an aqueous ethanol solution, 0.1 to 0.2 parts by weight of EDTA, and 0.2 to 1 parts by weight of a glycol additive. The selected material is selected from one or more of 5 to 20 parts by weight of gelatin or photocurable protein, 1 to 100 parts by weight of a natural cellulose polymer material, or 1 to 100 parts by weight of synthetic acrylic polymer material beads.

[0121] With the second embodiment of the present invention as described above, physical properties depending on the constituent ratio were studied, and while changing the mixing ratio of the individual components, the influence of the individual components on the physical properties of the entire composition was studied.

[0122] Table 13 shows the experimental conditions in which only the mixing ratio of the ethanol aqueous solution was changed and the mixing ratios of the other components were fixed in the second embodiment of the present invention.

[0123] [Table 13]

[0124]

[0125] Table 14 shows experimental conditions in which only the mixing ratio of EDTA was changed and the mixing ratios of other components were fixed in the second embodiment of the present invention.

[0126] [Table 14]

[0127]

[0128] Table 15 shows experimental conditions in which only the mixing ratio of the glycol additive was changed and the mixing ratios of the other components were fixed in the second embodiment of the present invention.

[0129] [Table 15]

[0130]

[0131] Table 16 shows experimental conditions in which only the mixing ratio of gelatin was changed and the mixing ratios of other components were fixed in the second embodiment of the present invention.

[0132] [Table 16]

[0133]

[0134] In Tables 13 to 16, the gelled cell fixation means exhibited good fixation capabilities in the Examples (A13 to A16, B13 to B16, and C13 to C16). On the other hand, the Comparative Examples (a13 to a16 and b13 to b16) exhibited either excessively strong or weak fixation capabilities for the sample within the vial 10. Consequently, in the Comparative Examples, problems (e.g., sample deformation) occurred during the sample removal process (e.g., re-dissolution) from the vial 10.

[0135] Regarding the second embodiment of the present invention, experiments other than those shown in Tables 13 to 16 were performed, but descriptions of the other experiments are omitted for convenience of explanation.

[0136] At the same time, in a second embodiment of the present invention, 5 to 20 parts by weight of any one or more of gelatin or photocurable protein can be replaced with 1 to 100 parts by weight of a natural cellulose polymer (e.g., hydroxypropyl methylcellulose (mecellose) or hydroxyethyl cellulose (hecellose)) or 1 to 100 parts by weight of a synthetic acrylic polymer bead (e.g., methyl methacrylate-co-ethylene glycol dimethacrylate; MMA).

[0137] Here, also for the natural cellulose polymer, experiments were conducted with the mixing ratio of the natural cellulose polymer ranging from less than 1 part by weight to greater than 100 parts by weight as a comparative example, while the mixing ratios of other components were fixed. The result was that in the range outside 1 to 100 parts by weight, the ability to fix the sample inside the vial 10 was poor.

[0138] In addition, for the synthetic acrylic polymer beads, experiments were conducted as comparative examples with mixing ratios of the synthetic acrylic polymer beads ranging from less than 1 part by weight to greater than 100 parts by weight, while the mixing ratios of the other components were fixed. As a result, the ability to fix the sample inside the vial 10 was poor in the range outside the range of 1 to 100 parts by weight.

[0139] Meanwhile, the redissolution solution used in step S240 in the second embodiment of the present invention may be configured to include 30 to 60 parts by weight of a methanol aqueous solution, 0.1 to 0.2 parts by weight of EDTA, 0.01 to 0.05 parts by weight of bile acid, and 0.05 to 0.1 parts by weight of a 1N acetic acid aqueous solution (hereinafter referred to as the "third redissolution solution").

[0140] In the second embodiment of the present invention, physical properties depending on the constituent ratio of the third re-dissolution solution were studied, and the influence of the individual components on the physical properties of the entire composition was studied while changing the mixing ratio of the individual components.

[0141] Table 17 shows experimental conditions in which only the mixing ratio of the methanol aqueous solution was changed and the mixing ratios of the other components were fixed during the redissolution process using the third redissolution solution in the second embodiment of the present invention.

[0142] [Table 17]

[0143]

[0144] Table 18 shows the experimental conditions in which only the mixing ratio of EDTA was changed and the mixing ratios of other components were fixed during the re-dissolution process using the third re-dissolution solution in the second embodiment of the present invention.

[0145] [Table 18]

[0146]

[0147] Table 19 shows the experimental conditions in which only the mixing ratio of bile acid was changed and the mixing ratios of other components were fixed during the reconstitution process using the third reconstitution solution in the second embodiment of the present invention.

[0148] [Table 19]

[0149]

[0150] Table 20 shows experimental conditions in which only the mixing ratio of the 1N acetic acid aqueous solution was changed and the mixing ratios of the other components were fixed during the redissolution process using the third redissolution solution in the second embodiment of the present invention.

[0151] [Table 20]

[0152]

[0153] At this time, in Tables 17 to 20, in Examples (A17 to A20, B17 to B20, and C17 to C20), there were no particular problems in the process of removing the sample from the vial 10 (e.g., re-dissolution). On the other hand, in Comparative Examples (a17 to a20 and b17 to b20), problems (e.g., deformation of the sample) occurred in the process of removing the sample from the vial 10 (e.g., re-dissolution).

[0154] Meanwhile, the redissolution solution in the second embodiment of the present invention may be configured to include 40 to 50 parts by weight of an ethanol aqueous solution, 0.1 to 0.2 parts by weight of EDTA, and 0.2 to 1 parts by weight of a glycol additive (hereinafter referred to as "the fourth redissolution solution").

[0155] In the second embodiment of the present invention, physical properties depending on the constituent ratio of the fourth redissolution solution were studied, and the influence of the individual components on the physical properties of the entire composition was studied while changing the mixing ratio of the individual components.

[0156] Table 21 shows the experimental conditions in which only the mixing ratio of the ethanol aqueous solution was changed and the mixing ratios of the other components were fixed during the re-dissolution process using the fourth re-dissolution solution in the second embodiment of the present invention.

[0157] [Table 21]

[0158]

[0159] Table 22 shows the experimental conditions in which only the mixing ratio of EDTA was changed and the mixing ratios of other components were fixed during the re-dissolution process using the fourth re-dissolution solution in the second embodiment of the present invention.

[0160] [Table 22]

[0161]

[0162] Table 23 shows experimental conditions in which only the mixing ratio of the glycol additive was changed and the mixing ratios of the other components were fixed during the redissolution process using the fourth redissolution solution in the second embodiment of the present invention.

[0163] [Table 23]

[0164]

[0165] In Tables 21 to 23, in Examples (A21 to A23, B21 to B23, and C21 to C23), there were no particular problems in the process of removing the sample from the vial 10 (e.g., re-dissolution). On the other hand, in Comparative Examples (a21 to a23 and b21 to b23), problems (e.g., deformation of the sample) occurred in the process of removing the sample from the vial 10 (e.g., re-dissolution).

Claims

1. A cell examination method using gelation of an alcohol solution composition, the method comprising: (a) mixing any one of a first solution comprising 30 to 60 parts by weight of a methanol aqueous solution, 0.1 to 0.2 parts by weight of EDTA, 0.01 to 0.05 parts by weight of bile acid, and 0.05 to 0.1 parts by weight of a 1N acetic acid aqueous solution or a second solution comprising 40 to 50 parts by weight of an ethanol aqueous solution, 0.1 to 0.2 parts by weight of EDTA, and 0.2 to 1 parts by weight of a glycol additive with 5 to 20 parts by weight of gelatin at a temperature of 1° C. to 40° C. for gelation to produce an alcohol-based jelly-like material; (b) placing the collected sample into a sealed container containing the alcohol jelly material; (c) preparing a redissolution solution comprising 30 to 60 parts by weight of an aqueous methanol solution, 0.1 to 0.2 parts by weight of EDTA, 0.01 to 0.05 parts by weight of cholic acid, and 0.05 to 0.1 parts by weight of a 1N acetic acid aqueous solution; (d) adding the gelled alcohol jelly material in the sealed container to the re-dissolution solution; (e) maintaining the re-dissolving solution at a temperature of 1° C. to 40° C. to re-dissolve the alcoholic jelly-like material into a liquid phase; and (f) Obtaining a sample from the alcohol jelly-like material in a reconstituted state and examining the sample.

2. A cell examination method using gelation of an alcohol solution composition, the method comprising: (a) placing any one of a first solution comprising 30 to 60 parts by weight of a methanol aqueous solution, 0.1 to 0.2 parts by weight of EDTA, 0.01 to 0.05 parts by weight of cholic acid, and 0.05 to 0.1 parts by weight of a 1N acetic acid aqueous solution, or a second solution comprising 40 to 50 parts by weight of an ethanol aqueous solution, 0.1 to 0.2 parts by weight of EDTA, and 0.2 to 1 parts by weight of a glycol additive into a sealed container, hereinafter referred to as the "selected solution"; (b) placing the sample into a sealed container filled with the selected solution; (c) placing 5 to 20 parts by weight of gelatin into a sealed container containing the selected solution and the sample at a temperature of 1° C. to 40° C. and gelling the mixture into an alcoholic jelly-like material; (d) preparing a redissolution solution comprising 30 to 60 parts by weight of an aqueous methanol solution, 0.1 to 0.2 parts by weight of EDTA, 0.01 to 0.05 parts by weight of cholic acid, and 0.05 to 0.1 parts by weight of a 1N acetic acid aqueous solution; (e) adding the gelled alcohol jelly material in the sealed container to the re-dissolution solution; (f) maintaining the re-dissolving solution at a temperature of 1° C. to 40° C. to re-dissolve the gelled alcohol-based jelly material into a liquid material; and (g) Obtaining a sample from the reconstituted liquid material and examining the sample.

3. A cell examination method using gelation of an alcohol solution composition, the method comprising: (a) mixing any one of a first solution comprising 30 to 60 parts by weight of a methanol aqueous solution, 0.1 to 0.2 parts by weight of EDTA, 0.01 to 0.05 parts by weight of bile acid, and 0.05 to 0.1 parts by weight of a 1N acetic acid aqueous solution or a second solution comprising 40 to 50 parts by weight of an ethanol aqueous solution, 0.1 to 0.2 parts by weight of EDTA, and 0.2 to 1 parts by weight of a glycol additive with 5 to 20 parts by weight of gelatin at a temperature of 1° C. to 40° C. for gelation to produce an alcohol-based jelly-like material; (b) placing the collected sample into a sealed container containing the alcohol jelly material; (c) preparing a redissolution solution comprising 40 to 50 parts by weight of an ethanol aqueous solution, 0.1 to 0.2 parts by weight of EDTA, and 0.2 to 1 parts by weight of a glycol additive; (d) adding the gelled alcohol jelly material in the sealed container to the re-dissolution solution; (e) maintaining the re-dissolving solution at a temperature of 1° C. to 40° C. to re-dissolve the alcoholic jelly-like material into a liquid phase; and (f) Obtaining a sample from the alcohol jelly-like material in a reconstituted state and examining the sample.

4. A cell examination method using gelation of an alcohol solution composition, the method comprising: (a) placing any one of a first solution comprising 30 to 60 parts by weight of a methanol aqueous solution, 0.1 to 0.2 parts by weight of EDTA, 0.01 to 0.05 parts by weight of cholic acid, and 0.05 to 0.1 parts by weight of a 1N acetic acid aqueous solution, or a second solution comprising 40 to 50 parts by weight of an ethanol aqueous solution, 0.1 to 0.2 parts by weight of EDTA, and 0.2 to 1 parts by weight of a glycol additive into a sealed container, hereinafter referred to as the "selected solution"; (b) placing the sample into a sealed container filled with the selected solution; (c) placing 5 to 20 parts by weight of gelatin into a sealed container containing the selected solution and the sample at a temperature of 1° C. to 40° C. and gelling the mixture into an alcoholic jelly-like material; (d) preparing a redissolution solution comprising 40 to 50 parts by weight of an ethanol aqueous solution, 0.1 to 0.2 parts by weight of EDTA, and 0.2 to 1 parts by weight of a glycol additive; (e) adding the gelled alcohol jelly material in the sealed container to the re-dissolution solution; (f) maintaining the re-dissolving solution at a temperature of 1° C. to 40° C. to re-dissolve the gelled alcohol-based jelly material into a liquid material; and (g) Obtaining a sample from the reconstituted liquid material and examining the sample.

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