JP2014163714A - Evaporation prevention membrane - Google Patents

Abstract

PROBLEM TO BE SOLVED: To prevent leakage and evaporation of a solution in a sample plate even tilting the sample plate by any angle and even when inserting or removing a capillary cathode end vertically to the sample plate in this state.SOLUTION: A septum is shaped with a taper and a lid pressing the septum is also shaped with a similar taper so as to make an external force always applied toward a capillary cathode end insertion part of the septum. This structure enables a hole opened by an insertion to be shield even inserting or removing the capillary cathode end. The outer periphery of the septum has a structure of an O ring which allows a gap between a container and the septum to be occluded by a crushing flange of the septum so as to prevent evaporation and leakage of a reagent in the container.

Description

  The present invention relates to an evaporation preventing film. For example, the present invention relates to a capillary electrophoresis apparatus that separates and analyzes materials such as DNA and proteins by electrophoresis, and particularly relates to a solution storage unit that stores solutions such as a material solution, a buffer solution, a reagent solution, and a washing solution.

  In recent years, capillary electrophoresis apparatuses in which capillaries are filled with an electrophoresis medium such as a polymer gel or a polymer solution have been widely used.

  For example, a capillary electrophoresis apparatus as disclosed in Patent Document 1 has been conventionally used. Compared with a flat-plate type electrophoresis apparatus, the heat release is high, and a higher voltage can be applied to the sample. Therefore, there is an advantage that electrophoresis can be performed at high speed. In addition, it has many advantages such as a small amount of sample, automatic filling of separation media, and automatic sample injection, and it is used for various separation analysis measurements including DNA and protein analysis.

  This DNA sequencer includes a mechanism that can automatically fill the capillaries with a syringe or pump and introduce the sample into the capillaries, etc., allowing continuous sample separation and analysis even in unattended conditions such as overnight operation. Measurements can be made.

  By the way, a sample to be electrophoresed is usually dispensed into a container in a state dissolved in a solvent. For example, in the case of performing DNA separation analysis, the DNA to be analyzed is amplified in advance by a technique such as PCR. For this reason, the sample plate for thermal cycler and the sample tube for PCR used for the amplification reaction of DNA by this PCR apparatus are often used as they are as a sample container when the purified DNA is separated and analyzed with a capillary electrophoresis apparatus.

  This eliminates the need to transfer the sample to a separate container for measurement using a capillary electrophoresis device for the sample that has been amplified and purified by the PCR device. Can be convenient. At this time, in order to prevent evaporation of the reagent, the upper surface side of the sample plate is covered with a rubber sheet called a septa as shown in Patent Document 2. The notch is formed in the septa, and when the sample introduction end of the capillary is inserted into the septa, the notch is expanded by pressing and can be inserted into the sample plate. Except when the sample introduction end of the capillary is inserted, the notch of the septa is closed, so that evaporation of the sample in the sample plate can be prevented. Further, this notch allows the sample or the like attached to the sample introduction end of the capillary to be wiped off when the sample introduction end of the capillary is withdrawn from the sample plate.

Japanese Patent No. 2776208 JP 2009-42226

  In order to reduce the size of the apparatus, the arrangement of the capillaries is important. Since the capillary cathode end is always inserted perpendicularly to the sample plate, the angle at which the sample plate can be installed is widened, so that the degree of freedom of capillary arrangement is increased.

  In an electrophoresis apparatus using a current capillary, the sample plate is installed horizontally, that is, the capillary cathode end is installed so as to face downward in the vertical direction. Therefore, the septa for preventing the evaporation of the reagent in the sample plate is assumed only in this case, and when the sample plate is inserted into the sample plate other than the conventional horizontal installation, that is, the capillary cathode end is not vertically downward. This makes it difficult to prevent reagent leakage and evaporation during insertion.

  The present invention employs the configurations described in the claims. One aspect is as follows.

  Separators that can prevent reagent leakage and evaporation when the capillary cathode end is inserted even if the sample plate is installed at any angle do not have a conventional notch, and when the capillary cathode end is inserted into this septa The sample and reagent in the sample plate are sucked up from the capillary cathode end by breaking through the septa formed of an elastic body. At this time, since the hole matching the shape of the capillary cathode end is opened in the septa, the capillary cathode end and the septa are easily sealed.

  In addition, an external force is always applied to the septa where the capillary cathode end is inserted, and the external force is opened when the capillary cathode end is inserted to work in a direction to close the hole opened when the capillary cathode end is inserted. The hole and the capillary cathode end are more closely attached. This is effective even when the capillary cathode end is removed from the septa, and the liquid is not leaked because the hole is closed by an external force.

  The sample plate container and septa are sealed with an O-ring structure on the outer periphery of the septa. The crushing allowance fills the gap and seals it, preventing reagent leakage and evaporation from between the container and septa. prevent.

According to the present invention, when the capillary cathode end is inserted into the sample plate other than vertically downward, it is possible to prevent reagent leakage and evaporation during insertion.

Diagram showing outline of capillary electrophoresis device Disassembled perspective view of sample plate Sample plate assembly drawing Cross section of sample plate The figure which shows the taper part of the scepter of a sample plate, and the structural example of a crushing allowance The figure which shows the taper part of the scepter of a sample plate, and the structural example of a crushing allowance The figure which shows the taper part of the scepter of a sample plate, and the structural example of a crushing allowance

  FIG. 1 shows an outline of an example of a capillary electrophoresis apparatus. The capillary electrophoresis apparatus of this example is a capillary array including one or a plurality of capillaries 101, a pump mechanism 103 for injecting the capillaries 101 into a polymer, and irradiating the sample in the capillary 101 with light to detect the fluorescence of the sample An optical system 104 for performing the operation, a high-voltage power source 105 for applying a high voltage to the capillary 101, an oven 106 for maintaining the capillary 101 at a constant temperature, and an autosampler 107 for transporting a container containing a sample, a solution, and the like.

  The capillary 101 is a replaceable member, and is replaced when the measurement technique is changed or when the capillary 101 is damaged or deteriorated in quality. The capillary 101 is composed of a glass tube having an inner diameter of several tens to several hundreds of microns and an outer diameter of several hundreds of microns, and the surface is coated with polyimide. The inside of the capillary 101 is filled with a separation medium for giving a migration speed difference during electrophoresis. The separation medium has both fluidity and non-fluidity, but in this embodiment, a fluid polymer is used.

  A capillary head 116 is provided at one end of the capillary 101, and a capillary cathode side 117 is formed at the other end. The capillary head 116 is a bundle of ends of the capillary 101 and has a function of connecting the pump mechanism 103 and the capillary 101. The capillary cathode end 117 is in contact with a sample, a solution, or the like. The capillary 101 is fixed by a load header 102 on the capillary cathode end side. The load header 102 is provided with a cathode electrode 114.

  The optical system 104 includes an irradiation system and a detection system. The optical system 104 has a function of irradiating excitation light to a portion of the capillary 101 where the polyimide coating is removed, that is, a detection unit. The detection system has a function of detecting fluorescence from the sample in the detection unit of the capillary 101. The sample is analyzed by the light detected by the detection system.

  The pump mechanism 103 includes a syringe 108, a block 109, a reverse support valve 110, a polymer container 111, and an anode buffer container 112. By connecting the capillary head 116 to the block 109, the capillary 101 and the flow path in the block 109 are connected. By operating the syringe 108, the polymer in the polymer container 111 is filled or refilled into the capillary 101 via the flow path in the block 109. The refilling of the polymer in the capillary 101 is performed for each measurement in order to improve the measurement performance. An anode electrode 113 is disposed in the anode buffer container 112. The high voltage power source 105 applies a high voltage between the anode electrode 113 and the cathode electrode 114.

  The oven 106 sandwiches the capillary 101 on a flat surface from a temperature control plate 115 to which a heat insulating material and a heater are attached, and keeps the capillary temperature constant. A temperature sensor for feedback is attached to the temperature control plate. Further, by fixing the load header 102 of the capillary array to the oven, the tip of the capillary head 116 can be fixed at a desired position.

  The autosampler 107 includes three electric motors and a linear guide for moving the moving stage, and the moving stage can be moved in three axial directions of up and down, left and right, and depth. The moving stage can transport the buffer container, the cleaning container, the waste liquid container, and the sample plate 118 to the capillary cathode end 117 (capillary electrode) as necessary.

  With reference to FIG. 2A, FIG. 2B, and FIG. 2C, the configuration of the sample plate 118 of this example used by the capillary electrophoresis apparatus will be described. As shown in FIG. 2A, the sample plate 118 of this example includes a container 203, a septa 202, and a lid 201.

  The septa 202 is fixed by being sandwiched between the container 203 and the lid 201, and the septa 202 may be formed independently or integrally with the lid 201 by insert molding. A seal structure between the lid 201 of the septa 202 and the container 203 will be described later.

  FIG. 2B shows a state where the sample plate 118 is assembled. In this example, the container 203 and the lid 201 are joined by an adhesive or ultrasonic fusion.

  FIG. 2C shows the usage state of the sample plate 118 of this example. As illustrated, the container 203 stores a solution 301 such as a reagent solution, a buffer solution, a sample solution, and a cleaning solution. The capillary cathode end 117 passes through the septa 202.

  The structure of the septa 202 will be described with reference to FIGS. 3A, 3B, and 3C. As shown in FIG. 3A, the scepter 202 has a recess 205 so that the capillary cathode end 117 can be easily penetrated. Further, the septa 202 has a tapered portion 203 so that an external force is always applied toward the capillary cathode end 117 insertion portion, and the tapered portion 203 is always in contact with the tapered portion 206 of the lid 201 shown in FIG. 3B. An external force is always applied toward the capillary cathode end 117 insertion portion. Further, the outer periphery 204 of the septa 202 has an O-ring structure, and when the lid is attached, the gap between the septa 202 and the container 203 is filled by the crushing allowance 207 of the outer periphery 204 as shown in FIG. The structure can prevent evaporation and leakage.

As mentioned above, although the example of this invention was demonstrated, this invention is not limited to this, It is understood by those skilled in the art that various changes are possible in the range of the invention described in the claim. The It is also within the scope of the present invention to appropriately combine the embodiments.

101 ... Capillary
102: Road header
103 ・ ・ ・ Pneumatic control system
104 ... Optical system
105 ・ ・ ・ High voltage power supply
106 ... Oven
107 ・ ・ ・ Autosampler
108 ・ ・ ・ Syringe
109 ・ ・ ・ Block
110 ・ ・ ・ Reverse valve
111 ・ ・ ・ Polymer container
112 ・ ・ ・ Anode buffer container
113 ... Anode power supply
114 ・ ・ ・ Cathode power supply
115 ... Temperature control plate
116 ... Capillary head
117 ... Capillary cathode end
118 ... Sample plate
200 ... Lid
201 ・ ・ ・ Scepter
202 ・ ・ ・ Container
203 ・ ・ ・ Taper (Scepter)
204 ・ ・ ・ Outer circumference (scepter)
205 ... depression (scepter)
206 ・ ・ ・ Tapered part (lid)
207 ・ ・ ・ Crushing cost (scepter)
301 ・ ・ ・ Solution

Claims (6)

A recess through which the capillary cathode end passes,
A tapered portion formed around the depression and lowered as the distance from the depression increases;
With
When the capillary cathode end is inserted, the capillary cathode end passes through the recess, and the tapered portion is configured to apply an external force toward the insertion portion of the capillary cathode end,
Septa characterized by.   2. The scepter according to claim 1, wherein a material of the scepter is formed of a resin or a rubber material that is easily elastically deformed.   2. The scepter according to claim 1, wherein an O-ring structure is formed on an outer periphery of the tapered portion.   4. The scepter according to claim 3, wherein a crushing margin is formed in an O-ring structure on an outer periphery of the tapered portion.   2. The scepter according to claim 1, wherein the same number of septa surrounding the capillary cathode end as that of each capillary cathode end are used.   2. The scepter according to claim 1, wherein the scepter is applicable not only to a sample plate but also to a buffer container, a washing container, and the like.