JP2008216090A - Plate for centrifugal device, and centrifugal device - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a plate for a centrifugal device and the centrifugal device, capable of moving a liquid sample, by a centrifugal force to determine quantitatively the liquid sample in a sample holding part, and capable of making the liquid sample of fixed amount surely flow into a flow passage. <P>SOLUTION: In this plate 101 for moving the liquid sample by the centrifugal force after injecting the liquid sample into the plate having the sample holding part; a wall 109 for forming a quantitative determination part 110 for the quantitative determination, when centrifuged is provided in the sample holding part 107 to be covered along a circular center direction thereof; and the liquid sample is centrifuged at a rotational speed satisfying P<SB>1</SB>>P<SB>2</SB>, where P<SB>1</SB>represents the pressure required for making the liquid sample flow into the flow passage 108, and P<SB>2</SB>represents pressure applied to the flow passage 108, when centrifuged. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention quantifies a liquid sample for discriminating a biological sample by detecting a transport reaction obtained by moving a biological sample such as DNA, protein or the like in a buffering agent. The present invention relates to a centrifuge plate for filling a centrifuge and a centrifuge.

  When a general biological sample is considered, DNA and protein exist largely. In recent years, with the rapid development of molecular biology, the involvement of genes in various diseases has been understood fairly accurately, and attention has been focused on medical treatments targeting genes.

  With regard to DNA, SNPs (abbreviation of single nucleotide polymorphism, which is generally translated as “single nucleotide polymorphism” and is a general term for the difference of one code (one base) in a gene) are currently attracting attention. The reason for this is that the classification of SNPs can predict the prevalence of many diseases and the effects and susceptibility of each individual to drugs, and there are absolutely no humans on the planet who have the same SNPs, even if they are parents and siblings. This is because it is considered that the individual can be completely identified from not doing so.

  Currently, as a method for examining SNPs, sequencing (determination of base sequence) in which the base sequence of DNA is directly read from the end is most commonly used. There are several reports on the sequencing method, but the most common method is dideoxy sequencing (Sanger method). Sequencing is based on a technique that separates and identifies the difference in length of one base length by denaturing polyacrylamide gel electrophoresis with high resolution or capillary electrophoresis in any method including the Sanger method. This is true.

Another method is affinity ligand capillary electrophoresis.
Affinity ligand capillary electrophoresis uses intermolecular affinity, particularly specific affinity in the ecosystem (enzyme and substrate, antigen and antibody affinity, etc.) to give specificity to separation. Specifically, When an affinity ligand that specifically recognizes the base sequence is added to the electrophoresis solution in the capillary tube and the sample is electrophoresed, only the molecular species that interact in the sample mixture will change in the moving speed. The analysis is performed with attention (for example, see Patent Document 1).

  On the other hand, proteins exist in cells, tissues, and biological fluids, regulate biological activities, supply energy to cells, synthesize important substances, maintain biological structures, and communicate between cells and intracellular information. Involved in transmission. Currently, it has become clear that proteins have multiple functions depending on the various environments, the presence of other interacting proteins, and the degree and type of modification that the protein has undergone.

  Proteins are made by connecting 20 types of amino acids in order according to gene instructions (sequence information), and the types are said to be tens of millions. If you know the sequence of the gene, what amino acid is what? You can get information on whether they are connected in order. A complete set of proteins made from the genes (genomes) of organisms is called a proteome, and now the analysis of the proteome has been actively promoted after the nucleotide sequence of the human genome has been deciphered.

Such functional analysis of proteins requires not only identification and characterization, but also biochemical assays, protein-protein interaction studies, protein networks, and intracellular and extracellular signaling elucidation. Research on this protein function uses a variety of techniques, including enzyme assays, yeast two-hybrid assays, chromatographic purification, information tools and databases, etc. Protein discrimination by electrophoresis is particularly important. It is a technique. Then, as in electrophoresis, the transport reaction obtained when moving liquids such as samples, analytes, buffers, and reagents in capillary tubes is detected, and analysis, discrimination, determination, etc. of the samples are performed. There are various reports regarding the transportation and orientation of the liquid when it is performed (for example, Patent Documents 2 to 4).
JP 7-311198 A Special Table 2000-513813 JP-T-2001-523341 International Publication No. 2005/064339 Pamphlet

  Conventionally, when a liquid sample for detecting a transport reaction obtained by moving a DNA, protein or other biological sample in a buffer to detect the biological sample is centrifuged, the liquid sample is placed on a plate for a centrifuge, etc. However, since the amount of liquid sample to be injected varies, it is difficult to reliably fill the flow channel with a certain amount, and the amount of liquid sample filled in the flow channel is excessive or insufficient. There's a problem.

  Further, in the method described in Patent Document 4, the liquid sample is moved by embedding a fine channel in the platform and changing the rotational speed of the platform to change the centripetal force resulting from the rotation. Then, since the liquid sample flows into the flow path by the first centrifugation, there is a problem that the flow path cannot be used when the liquid sample is forgotten or insufficient.

  The present invention has been made in order to solve the above-described problems. The liquid sample is moved by centrifugal force, the liquid sample is quantified in the sample holding unit, and a certain amount of liquid sample is surely passed through the flow path. It aims at providing the plate for centrifuges which can be filled in, and the centrifuge.

  The centrifuge device plate according to claim 1 of the present invention is the centrifuge device plate in which the liquid sample is moved by centrifugal force after injecting the liquid sample into the sample holder, and the sample holder is centrifuged. In this case, a quantification unit for quantifying the liquid sample is formed by a wall projecting in the direction of the center from the side wall of the outermost peripheral part of the sample holding part, and the plate from the outermost peripheral part of the quantification part of the sample holding part A flow path extending in the centrifugal direction is provided, and the liquid sample is injected into a position on the inner peripheral side of the plate with respect to the position of the wall in the sample holding portion.

According to a second aspect of the present invention, there is provided a centrifugal apparatus using the centrifugal apparatus plate according to the _{first aspect} , wherein the pressure required to flow the liquid sample into the flow path is P ₁ , and the pressure is obtained by centrifugation. If the pressure on the flow path was P _2, characterized in that centrifuging at a rotational speed to be P _1> P _2.

A centrifugal device according to a third aspect of the present invention is the centrifugal device according to the second aspect, wherein the pressure required to flow the liquid sample into the flow path is P ₁ , and the pressure applied to the flow path by centrifugation is set. When P ₂ is set, the centrifugation is temporarily stopped after the rotation speed satisfying P ₁ > P _2, and then the centrifugation is performed again at the rotation speed satisfying P ₁ <P ₂ .

  The centrifuge device plate according to claim 4 of the present invention is the centrifuge device plate according to claim 1, wherein the wall protrudes so as to cover the sample holding portion in the center direction and is injected. Is characterized in that the liquid sample to be quantified by being centrifuged is moved to the lower part of the wall, and the liquid sample not to be quantified is separated to the upper part of the wall.

  The centrifuge device plate according to claim 5 of the present invention is characterized in that, in the centrifuge device plate according to claim 1, the flow path is formed in a fixed portion of the lower part of the wall.

The centrifuge device plate according to claim 6 of the present invention is the centrifuge device plate according to claim 1, wherein the liquid sample is located at a position in the center of gravity direction of the plate with respect to the sample holding portion of the centrifuge device plate. A sample injection part for injecting the liquid sample is provided, and the pressure required to flow the liquid sample into the first flow path extending in the centrifugal direction of the plate from the outermost peripheral part of the sample holding part is P ₁ , and the _first pressure is obtained by centrifugation. When the pressure applied to one flow path is P ₂ , the second flow path through which the liquid sample flows into the sample holding section when centrifuged at a rotational speed satisfying P ₁ > P ₂ is defined as the sample injection section. The sample holder is provided with an inlet through which the liquid sample flows into the sample holder from the second flow path, and the inlet is provided with a centrifuge. When the liquid sample flows into the quantification unit In addition, one or a plurality of walls are provided at a position in the centrifugal direction of the plate so as to divide the sample holding part into left and right, and a liquid sample to be quantified is formed in the first flow path by centrifugation. The liquid sample that is moved to the outermost peripheral portion of the sample holding portion and separated is separated into a side on which the liquid sample to be quantified moves and an opposite side across the wall.

  The centrifuge device plate according to claim 7 of the present invention is the centrifuge device plate according to any one of claims 4 to 6, wherein the wall is easily wetted by the liquid sample or the liquid sample. It is the material which processed.

  The centrifuge device plate according to claim 8 of the present invention is the centrifuge device plate according to claim 7, wherein one or more pairs of positive and negative electrodes for confirming the presence or absence of the liquid sample based on the current value are provided. And provided in the quantification unit.

  The centrifuge device plate according to claim 9 of the present invention is the centrifuge device plate according to claim 8, wherein the liquid sample is held in an amount necessary to flow into the flow path based on a current value. A pair of positive and negative electrodes to be confirmed is provided on the innermost circumference of the wall.

  A centrifugal device plate according to a tenth aspect of the present invention is the centrifugal device plate according to the seventh aspect, wherein a plurality of pairs of positive electrodes and negative electrodes measure the amount of the liquid sample held based on a current value. Is provided from the innermost peripheral part to the outermost peripheral part of the wall in the fixed amount part.

  A centrifuge plate according to an eleventh aspect of the present invention is the centrifuge device plate according to the seventh aspect, wherein a pair of positive electrode and negative electrode measure the amount of the liquid sample held based on a current value. Is provided from the innermost peripheral part to the outermost peripheral part of the wall in the fixed amount part.

According to the centrifugal apparatus plate and the centrifugal apparatus of the present invention, a plurality of patterns are formed radially on the plate, and after the liquid sample is injected into the sample holder, the liquid sample is moved by centrifugal force. In the apparatus plate, the sample holding part forms a quantification part for quantifying the liquid sample when it is centrifuged with a wall protruding in a circular direction from a side wall of the outermost peripheral part of the sample holding part, A flow path extending in the centrifugal direction of the plate from the outermost peripheral portion of the quantification unit of the sample holding unit is provided, and the liquid sample is injected into a position in the center of gravity direction of the plate rather than the position of the wall in the sample holding unit. When the pressure required to flow the liquid sample into the flow path is P ₁ and the pressure applied to the flow path by centrifugation is P ₂ , centrifugation is performed at a rotational speed that satisfies P ₁ > P _2. By being injected The liquid sample can be quantified regardless of the amount of the liquid sample.

Further, according to the centrifuge device and the centrifuge of the present invention, when the pressure required to flow the liquid sample into the flow path is P ₁ and the pressure applied to the flow path by centrifugation is P ₂ , P Quantification of the liquid sample regardless of the amount of the injected liquid sample by centrifuging at a rotational speed satisfying ₁ > P ₂ and then suspending again at a rotational speed satisfying P ₁ <P ₂ Thus, it is possible to reliably fill the flow path with a certain amount of liquid sample.

  Moreover, according to the plate for centrifuges and the centrifuge of the present invention, the presence and amount of the liquid sample can be confirmed by providing electrodes in the quantification unit and measuring the current value.

(Embodiment 1)
Hereinafter, the centrifuge device plate and the centrifuge according to the first embodiment of the present invention will be described with reference to FIGS.
In Embodiment 1 of the present invention, the centrifuge device plate quantifies the injected liquid sample by the preliminary rotation, and the liquid sample flows into the flow path by the main rotation.

First, the configuration of the centrifuge device plate according to the first embodiment of the present invention will be described.
FIG. 1 is a view showing a flow path forming surface of a centrifuge device plate according to Embodiment 1 of the present invention.
In FIG. 1, eight patterns 102 are radially formed on the plate 101 according to the first embodiment.

  As shown in FIG. 1, the outer shape of the plate 101 in the first embodiment is a square of 8 centimeters, and three of the four corners are provided with rounded corners (R) so as to form part of a circle. The remaining corner is chamfered.

  The plate 101 is further provided with a hole 104, which has an asymmetric outer shape so that the location of the pattern can be specified. The plate 101 is made of acrylic resin and has a thickness of 2 mm. Grooves are formed on the flow path forming surface, and a sealed flow path is formed by adhering an acrylic film having a thickness of 50 μm. The center of gravity 105 is the center of gravity of the main plate, and a rotating portion fixing hole 103 for fixing the main plate to the rotating portion around the center of gravity 105 is provided.

Next, the pattern 102 will be described. For example, the pattern 102 in FIG. 1 has a configuration in which two sample holders 107 are connected by a tube.
FIG. 2 is a diagram showing a detailed shape of the pattern 102 formed on the plate 101 shown in FIG. FIG. 3 is a cross-sectional view of the pattern 102 taken along the line AA.

  The pattern 102 is provided with a sample inlet 106 through which a liquid sample is injected, a sample holder 107 for temporarily holding the injected sample, and a flow path 108 extending from the outermost periphery of the sample holder 107 in the outer peripheral direction. It has been.

  As shown in FIG. 3, the sample holding unit 107 has a wall 109 that has been processed so as to easily wet the liquid sample protruding from the outermost side wall of the sample holding unit 107 so as to cover the sample in the center direction. The portion 107 is divided into an upper portion and a lower portion of the wall 109. The flow path 108 is formed from the lower part of the wall 109, and the quantification unit 110 protrudes in the direction of the center from the side wall of the outermost peripheral part of the sample holding unit 107 that quantifies the liquid sample when centrifuged. It is the lower part of the wall 109. Here, the wall 109 may be made of a material that easily wets the liquid sample.

  Further, a positive electrode 111 and a negative electrode 112 are provided in the quantification unit 110. The positive electrode 111 is connected to the positive voltage application unit 113 and the negative electrode 112 is connected to the negative voltage application unit 114. By applying a positive voltage to the positive voltage application unit 113 and a negative voltage to the negative voltage application unit 114, The presence and amount of the liquid sample can be confirmed. Here, when the positive voltage application unit 113 and the negative voltage application unit 114 are provided on the flow path forming surface, the positive electrode 111 and the negative electrode 112 are provided on the plate forming the wall 109, and the positive voltage application unit 113 and the negative voltage application unit When 114 is provided on the back surface, the positive electrode 111 and the negative electrode 112 are provided on a film forming a closed flow path.

  Next, the pattern of the positive electrode 111 and the negative electrode 112 provided on the plate 101 according to the first embodiment of the present invention will be described.

  In the pattern 102 of the first embodiment, a plurality of pairs of electrodes are provided from the innermost peripheral portion to the outermost peripheral portion of the wall 109 including the innermost peripheral portion of the wall 109. In this case, since the number of electrodes through which the current flows changes depending on the amount of the liquid sample held in the sample holding unit 107, the presence and amount of the sample can be confirmed by applying a voltage and measuring the current value. Can do.

  Next, the configuration of the centrifugal apparatus according to the first embodiment of the present invention will be described with reference to FIG. FIG. 4 is a configuration diagram of the centrifuge according to the first embodiment of the present invention.

  The centrifuge according to the first embodiment has a motor 10 for moving a liquid sample in the plate 101, a plate tray 20 that can be rotationally driven by which the plate 101 can be mounted by the motor 10, and voltage application to the plate 101. And a control board 40 for controlling the operation of the centrifugal device 100.

  The voltage application means 30 includes electrode probes 31a and 31b for confirming the presence and amount of the liquid sample by the current value. Here, the electrode probes 31a and 31b are provided in the number of electrode pairs when a plurality of pairs of electrodes are provided on the plate 101, respectively. In the first embodiment, since eight patterns 102 are provided on the plate 101, eight electrode probes 31a and 31b are provided. This is not the case when more patterns 102 are provided.

  In the first embodiment, the voltage application means 30 is provided on the upper side of the tray 20. However, depending on the surface on which the positive voltage application unit 113 and the negative voltage application unit 114 are provided, for example, the wall shown in FIG. When the positive voltage application unit 113 and the negative voltage application unit 114 are provided on the lower surface of 109, the voltage application means 30 may be provided on the lower side of the tray 20.

  On the voltage application means base plate 32 which is the base of the voltage application means 30, an electrode substrate unit 33 to which electrode probes 31 a and 31 b for applying a voltage to the plate 101 are electrically connected and fixed is attached to the plate 101. In order to perform the separation or contact operation with respect to the electrode probe, it is arranged in a state of being engaged with an electrode driving means 34 such as a DC motor, but an electrode probe such as a compression spring is provided at the tip of the electrode probes 31a and 31b. By providing the built-in spring 35, the contact pressure of the electrode probes 31a and 31b with respect to the plate 101 is adjusted.

  Further, the centrifugal device 100 includes a power supply 50 connected to the electrode probes 31a and 31b of the voltage application means 30, a device power supply 60, a power switch 70 for switching the device on and off, and the power switch is in an ON state. LED 71 that is turned on at the time of cooling, a cooling fan 80 that cools the inside of the centrifuge, and a rubber leg 90 that can protect the centrifuge 100 from vibration and can be adjusted in height are provided.

Next, an example of specific operations and operations until the liquid sample is quantified will be described with reference to FIGS.
5 and 6 are diagrams showing the state of the liquid sample when the centrifuge device pattern 102 according to the first embodiment of the present invention is centrifuged.

  First, a liquid sample is injected into the plate, and preliminary rotation is performed to quantify the liquid sample.

The liquid sample is injected from the sample agent injection port 106 into the sample holding unit 107 by a pipetter or the like.

Next, the plate 101 is fixed to the plate tray 20 of the centrifuge 100, and the plate 101 is rotated about the center of gravity 105. At this time, the number of rotations is P ₁ > P ₂ when P _{1 is} the pressure required to allow the liquid sample to flow into the flow path 108 and P ₂ is the pressure applied to the flow path 108 when centrifugation is performed. Centrifugation is performed at a rotational speed at which the pressure required for the liquid sample to flow into the flow path 108 is greater than the pressure applied to the flow path 108 when centrifugation is performed.

  At this time, the liquid sample moves in the outer circumferential direction by centrifugal force, and the liquid surface is held on the same circumference in the liquid holding portion 107 as shown in FIG. Here, when the amount of the injected liquid sample is larger than the fixed amount, the liquid sample is moved to the lower part of the wall 109 by centrifugal force as shown in FIG. And quantified by the lower quantitative unit 110. Here, since the wall 109 is a material that easily wets the liquid sample, the DNA liquid sample can be easily separated.

  Next, preliminary rotation for quantifying the sample is temporarily stopped, and the amount of the liquid sample in the quantification unit 110 of the sample holding unit 107 is confirmed.

  The electrode probes 31 a and 31 b are pressed against the positive voltage application unit 113 and the negative voltage application unit 114 of the pattern 102 to apply a voltage.

  A plurality of pairs of the positive electrode 111 and the negative electrode 112 are provided from the innermost periphery of the wall 109 to the outermost periphery including the innermost periphery of the wall 109. Therefore, if a liquid sample is held in the quantification unit 110, a current flows through the electrode, and the presence or absence of the liquid sample can be confirmed by measuring the current value. Furthermore, by providing an electrode on the innermost periphery of the wall 109, it is possible to confirm the shortage of the liquid sample based on the presence or absence of an electric current of the electrode provided on the innermost periphery. Moreover, the shortage of the liquid sample can be confirmed by providing a plurality of electrodes and confirming the electrode through which the current flows.

  Here, the voltage to be applied is such that bubbles do not occur in the liquid sample due to the oxidation-reduction reaction.

If the amount of liquid sample is insufficient, an additional amount of liquid sample is injected. Then, the liquid sample is quantified by rotating again. Thereby, a certain amount of liquid sample can be reliably filled into the plate.

Next, the main rotation is performed in order to allow the liquid sample quantified in the preliminary rotation to flow into the channel 108.
Centrifuge again at a rotational speed at which P ₁ <P ₂ , that is, the pressure applied to the flow path 108 when centrifugation is greater than the pressure required to allow the liquid sample to flow into the flow path 108. By doing so, the quantified liquid sample flows into the channel 108.

  In the first embodiment, the case where the wall 109 is formed in the horizontal direction with respect to the plate surface has been described. However, the wall 109 may be formed in the vertical direction with respect to the plate surface.

  As described above, according to the first embodiment, the sample holder is provided with a wall for quantification when centrifuging, and the liquid sample is centrifuged at a rotational speed that does not flow into the flow path. By quantifying the liquid sample, a certain amount of the liquid sample can be flowed into the flow path. In addition, by providing the electrode, the presence and amount of the liquid sample can be confirmed, and the shortage can be replenished, so that there is no fear that the flow path becomes unusable.

(Embodiment 2)
Hereinafter, a plate for a centrifuge device and a centrifuge device according to Embodiment 2 of the present invention will be described with reference to FIGS.

  In the second embodiment of the present invention, another example of the pattern 102 formed on the plate 101 will be described using the same centrifugal device as in the first embodiment.

  FIG. 7 is a view seen from the flow path forming surface of the pattern 102 of the centrifuge device plate according to the second embodiment of the present invention.

  In the pattern 102, a sample injection port 115 into which a liquid sample is injected, a sample injection unit 116 for temporarily holding the injected liquid sample, a sample holding unit 117, and sample holding from the outermost periphery of the sample injection unit 116 A second flow path 118 extending to the portion 117 and a first flow path 119 extending from the outermost peripheral portion of the sample holding portion 117 in the outer peripheral direction are provided.

Here, the second flow path 118 has a pressure P ₁ necessary for flowing the liquid sample into the first flow path 119 and causes the liquid sample to flow into the first flow path 119 when centrifugation is performed. When the pressure necessary for the pressure is P ₂ , the rotation speed at which P ₁ > P ₂ , that is, the pressure necessary for the liquid sample to flow into the first flow path 119 is The liquid sample is provided so as to flow into the sample holding unit 117 when centrifugation is performed at a rotational speed that is greater than the pressure required to flow the liquid sample into one flow path 119.

  The sample holder 117 is provided with first and second walls 120a and 120b that have been processed so that the liquid sample is easily wetted, so that the left and right walls are divided in the direction of the center of the circle. The first channel 119 is formed by a portion sandwiched between the first and second walls 120a and 120b, and the first and second walls that quantify the liquid sample when the quantification unit 121 is centrifuged. It is a portion sandwiched between 120a and 120b. Here, the first and second walls 120a and 120b may be made of a material that easily wets the liquid sample.

  The inflow port 122 that flows into the sample holding unit 117 from the second flow path 118 is provided so that the liquid sample flows into the quantification unit 121 when it is centrifuged.

  The quantification unit 121 is provided with a positive electrode 123 and a negative electrode 124. The positive electrode 123 is connected to the positive voltage application unit 125, and the negative electrode 124 is connected to the negative voltage application unit 126. The positive voltage 123 is applied to the positive voltage application unit 125, and the negative voltage 124 is applied to the negative voltage application unit 126. Thus, the presence and amount of the liquid sample can be confirmed. Here, when the positive voltage application unit 125 and the negative voltage application unit 126 are provided on the flow path forming surface, the positive electrode 123 and the negative electrode 124 are provided on a plate that forms the back surface of the plate 101, and the positive voltage application unit 125 and the negative voltage application unit 126 are provided. When the application unit 126 is provided on the back surface, the positive electrode 123 and the negative electrode 124 are provided on a film that forms a sealed channel.

  Next, the pattern of the positive electrode 123 and the negative electrode 124 provided on the plate 101 will be described.

  In the pattern 102 according to the second embodiment of the present invention, a pair of electrodes is provided from the innermost periphery to the outermost periphery of the first and second walls 120a and 120b. In this case, since the resistance value varies depending on the amount of the liquid sample held in the sample holding unit 117, the presence and amount of the liquid sample can be confirmed by applying a voltage and measuring the current value.

Next, an example of specific operations and operations until the liquid sample is quantified in Embodiment 2 of the present invention will be described with reference to FIG.
FIG. 8 is a diagram showing the state of the liquid sample when the centrifuge device pattern 102 according to the second embodiment of the present invention is centrifuged.

  When the preparation of the liquid sample is completed, the liquid sample is injected into the plate, and preliminary rotation is performed to quantify the liquid sample.

  First, a liquid sample is injected into the plate. The liquid sample is injected from the liquid sample injection port 115 into the sample injection unit 116 by a pipetter or the like.

Next, when the plate 101 is fixed to the plate tray 20 of the centrifuge 100 as in the first embodiment, and the pressure necessary for flowing the liquid sample into the first flow path 119 is P ₁ and centrifugation is performed. first pressure on the flow path 119 in the case of the P _2, P _1> P ₂ become rotational speed, i.e., the pressure required to flow into the liquid sample into the first flow path 119, centrifugation Centrifuging at a rotation speed greater than the pressure applied to the first flow path 119.

At this time, since the second flow path 118 is provided so that the liquid sample flows into the sample holding portion 117 when being centrifuged at a rotation speed satisfying P ₁ > P ₂ , the liquid sample is The liquid sample moves in the outer circumferential direction, and moves from the inlet 122 to the quantification unit 121 of the sample holding unit 117 through the second flow path 118. As shown in FIG. 8, the liquid sample is held on the same circumference in the sample holder 117 as shown in FIG. At this time, if the injected liquid sample is larger than the volume of the quantification unit 121, the unnecessary liquid sample is separated to the left and right of the quantification unit 121 by the first and second walls 120a and 120b. Here, the first and second walls 120a and 120b are easily separated because they are easily wetted by the liquid sample.

  Next, preliminary rotation for quantifying the liquid sample is temporarily stopped, and the amount of the liquid sample in the quantification unit 121 of the sample holding unit 117 is confirmed.

  The first and second electrode probes 31a and 31b are pressed against the positive voltage application unit 125 and the negative voltage application unit 126 of the pattern 120 to apply a voltage.

  A pair of the positive electrode 123 and the negative electrode 124 is provided from the innermost periphery to the outermost periphery of the first and second walls 120a and 120b. Therefore, since the resistance value between the electrodes changes when the amount of the liquid sample changes, the presence and amount of the liquid sample and the sample can be confirmed by applying a voltage and measuring the current value.

  Here, the voltage to be applied is such that bubbles do not occur in the liquid sample due to the oxidation-reduction reaction. If the amount of liquid sample is insufficient, an additional amount of liquid sample is injected. Then, the liquid sample is quantified by rotating again. Thereby, a certain amount of liquid sample can be reliably filled into the plate.

Next, in order to flow the liquid sample quantified by the preliminary rotation into the first channel 119, the main rotation is performed.
Rotation speed at which P ₁ <P ₂ , that is, rotation at which the pressure applied to the first flow path 119 when centrifugation is performed is greater than the pressure required for the liquid sample to flow into the first flow path 119. Centrifugation is again performed with the number, and the quantified liquid sample flows into the first flow path 119.

  In the second embodiment, the case where the walls 120a and 120b are formed in the direction perpendicular to the plate surface has been described. However, the walls 120a and 120b may be formed in the horizontal direction with respect to the plate surface.

  As described above, according to the second embodiment, the sample holder is provided with a wall for quantification when centrifuging, and the liquid sample is centrifuged at a rotational speed that does not flow into the flow path. By quantifying the liquid sample, a certain amount of the liquid sample can be flowed into the flow path. In addition, by providing the electrode, the presence and amount of the liquid sample can be confirmed, and the shortage can be replenished, so that there is no fear that the flow path becomes unusable.

  The plate for a centrifuge and the centrifuge of the present invention are useful for reliably quantifying a liquid sample before filling the flow path.

The figure which shows the flow-path formation surface of the plate for centrifuges by Embodiment 1 of this invention. The figure which shows the pattern 102 formed in the plate for centrifuges by Embodiment 1 of this invention. The figure which shows the cross section of the pattern 102 formed in the plate for centrifuges by Embodiment 1 of this invention. The figure which shows the structure of the centrifuge by Embodiment 1 of this invention. The figure which shows the state of the liquid sample when centrifuging to the pattern 102 of the plate for centrifuges by Embodiment 1 of this invention. Plate sectional view which shows the state of the liquid sample when centrifuging to the pattern 102 of the plate for centrifuges by Embodiment 1 of this invention The figure which shows the pattern 102 formed in the plate for centrifuges by Embodiment 2 of this invention. The figure which shows the state of the liquid sample when centrifuging to the pattern 102 of the plate for centrifuges by Embodiment 2 of this invention.

Explanation of symbols

101 Plate 102 Flow path pattern 103 Rotating part fixing hole 104 Positioning hole 105 Plate center of gravity 106 Sample inlet 107 Sample holding part 108 Channel 109 Wall 110 Fixed part 111 Positive electrode 112 Negative electrode 113 Positive voltage applying part 114 Negative voltage applying part 115 Sample injection port 116 Sample injection unit 117 Sample holding unit 118, 119 Channel 120a, 120b Wall 121 Determination unit 122 Inflow port 123 Positive electrode 124 Negative electrode 125 Positive voltage application unit 126 Negative voltage application unit

Claims (11)

In the centrifuge plate that moves the liquid sample by centrifugal force after injecting the liquid sample into the sample holder,
The sample holding part forms a quantification part for quantifying the liquid sample when it is centrifuged with a wall protruding in a circular direction from a side wall of an outermost peripheral part of the sample holding part, and the sample holding part Provide a flow path extending in the centrifugal direction of the plate from the outermost peripheral portion of the quantitative portion,
The liquid sample is injected into a position on the inner peripheral side of the plate rather than the position of the wall in the sample holding unit.
A centrifuge plate characterized by the above. In the centrifuge using the centrifuge device plate according to claim 1,
When the pressure required to flow the liquid sample into the channel is P ₁ and the pressure applied to the channel by centrifugation is P ₂ , the sample is centrifuged at a rotational speed such that P ₁ > P ₂ .
A centrifugal device characterized by that. The centrifuge according to claim 2,
When the pressure required to flow the liquid sample into the flow path is P ₁ and the pressure applied to the flow path by centrifugation is P ₂ , the pressure is temporarily applied after the rotation at P ₁ > P _2. Stop and centrifuge again at a speed of P ₁ <P ₂ ,
A centrifugal device characterized by that. The centrifuge plate according to claim 1,
The wall protrudes so as to cover the sample holder in the direction of the center of the circle,
The injected liquid sample is centrifuged to move the liquid sample to be quantified to the lower part of the wall, and the liquid sample not to be quantified is separated to the upper part of the wall.
A centrifuge plate characterized by the above. The centrifuge plate according to claim 1,
The flow path is formed in the fixed portion at the bottom of the wall,
A centrifuge plate characterized by the above. The centrifuge plate according to claim 1,
A sample injection unit for injecting the liquid sample is provided at a position in the center of gravity of the plate with respect to the sample holding unit of the centrifuge device plate,
The pressure required to flow the liquid sample from the outermost peripheral portion of the sample holding portion into the first flow path extending in the centrifugal direction of the plate is P ₁ , and the pressure applied to the first flow path by centrifugation is P ₁ . ₂ , the second flow path through which the liquid sample flows into the sample holder when centrifuged at a rotational speed satisfying P ₁ > P ₂ is passed from the outermost peripheral part of the sample injection part to the sample holder. Set in a position that extends,
The sample holding unit is provided with an inlet through which the liquid sample flows into the sample holding unit from the second flow path, and the inlet flows into the quantification unit when the liquid sample is centrifuged,
One or a plurality of walls are provided at a position in the centrifugal direction of the plate so as to divide the sample holding part into left and right,
By centrifuging, the liquid sample to be quantified is moved to the outermost peripheral portion of the sample holding portion in which the first flow path is formed, and the liquid sample not to be quantified is moved to the side on which the liquid sample to be quantified is moved. Separate the opposite side across the wall,
A centrifuge plate characterized by the above. The centrifuge plate according to any one of claims 4 to 6,
The wall is a material that easily wets the liquid sample, or a material that has been processed to easily wet the liquid sample.
A centrifuge plate characterized by the above. The centrifuge plate according to claim 7,
One or more pairs of a positive electrode and a negative electrode for confirming the presence or absence of the liquid sample based on a current value are provided in the quantitative unit.
A centrifuge plate characterized by the above. The centrifuge plate according to claim 8, wherein
A pair of positive and negative electrodes is provided on the innermost circumference of the wall to check whether the amount of the liquid sample necessary to flow into the flow path is held based on a current value.
A centrifuge plate characterized by the above. The centrifuge plate according to claim 7,
A plurality of pairs of positive electrodes and negative electrodes for measuring the amount of the liquid sample held based on a current value are provided from the innermost peripheral part to the outermost peripheral part of the wall in the quantification part,
A centrifuge plate characterized by the above. The centrifuge plate according to claim 7,
A pair of positive and negative electrodes for measuring the amount of the liquid sample held based on the current value is provided from the innermost periphery to the outermost periphery of the wall in the quantification unit,
A centrifuge plate characterized by the above.

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