JP4512465B2 - Electrophoresis device - Google Patents

Description

  The present invention relates to a technique for separating and analyzing nucleic acids, proteins, and the like by electrophoresis, for example, a capillary array electrophoresis apparatus.

  2. Description of the Related Art In recent years, a capillary array type electrophoresis apparatus has been proposed that measures a number of capillaries filled with a gel in order to increase the speed and throughput when determining the base sequence of DNA. For example, Patent Document 1 discloses a sheath flow method in which detection is performed by using a plurality of capillaries and irradiating a laser in a buffer solution at the end of the capillary. Patent Document 2 discloses a capillary electrophoresis apparatus that has a capillary pre-filled with a gel and a buffer tank provided at the lower end of the capillary and performs fluorescence detection from the lower end surface of the capillary. Further, in Reference 3, the ends of the capillaries are arranged in a lattice pattern, and the spectrum-resolved detection image is collected on a two-dimensional detector using one or a plurality of spectroscopic elements (diffraction grating, prism). ing.

JP-A-8-193976 JP-A-10-19846 Japanese translation of PCT publication No. 2002-520616

  As a result of intensive studies by the inventor, when the ends of the capillaries are arranged in a lattice shape, the detection image of each capillary on the two-dimensional detector does not become a lattice shape, so that data acquisition on the two-dimensional detector is performed at high speed. I found it difficult.

  When the detection system using the spectroscopic element disclosed in Document 3 is used, the spectrum line is curved, and the image on the two-dimensional detector is curved toward the long wavelength side. In addition, since aberrations such as distortion occur due to the characteristics of the condenser lens, the emission spectrum of each capillary on the two-dimensional detector is not distorted and mapped in a lattice pattern. Since the emission spectrum of each capillary mapped on the two-dimensional detector is not arranged in a grid pattern, it is difficult to acquire data on the two-dimensional detector at high speed.

  An object of the present invention relates to high-sensitivity simultaneous measurement using a large number of capillaries and high-speed processing of measurement results.

  The present invention relates to adjusting an arrangement shape of a plurality of capillary ends and controlling image formation on the detector in an electrophoresis apparatus that detects light emitted from a plurality of capillary ends with a detector. Thereby, for example, an image can be formed in accordance with the arrangement of the optical elements in the detector, and data acquisition from the detector can be performed at high speed.

  Further, in the present invention, a plurality of capillary ends are fixed by a fixing member in an electrophoresis device in which light emitted from a plurality of capillary ends is detected by a detector and the electrophoresis medium is filled into the capillaries from the plurality of capillary ends. About doing. Thereby, for example, even when the electrophoresis medium is filled in the capillary, the arrangement accuracy of the plurality of capillary ends can be maintained, and the measurement sensitivity can be maintained.

  According to the present invention, high-sensitivity simultaneous measurement using a large number of capillaries and high-speed processing of measurement results are possible.

  The above and other novel features and benefits of the present invention will be described below with reference to the drawings. However, the drawings are for explanation only, and do not limit the scope of the present invention.

  FIG. 1 shows a schematic perspective view of the electrophoresis apparatus. FIG. 2 is a schematic perspective view of the capillary head. FIG. 5 shows a schematic sectional view in the vicinity of the polymer filling block. FIG. 6 shows a schematic flow diagram of the analysis procedure. The outline of the electrophoresis apparatus will be described below with reference to these drawings.

  The electrophoresis apparatus in the present embodiment (hereinafter referred to as “this apparatus”) uses a plurality of capillaries filled with an electrophoresis medium, introduces a sample into each capillary, and separates and analyzes the sample components in the sample. To do. For example, a base sequence can be analyzed by simultaneously analyzing a plurality of samples containing DNA. The basic configuration of this apparatus is a capillary array, an irradiation optical unit, a detection optical unit, an autosampler unit, an electrophoresis medium filling unit, a power supply unit, and a temperature control unit.

  The capillary array 101 is a detachable exchange member composed of a plurality of capillaries 102, and is exchanged for a new one when the quality deteriorates due to a predetermined number of analyzes and the separation performance decreases. When the measurement technique is changed, the length of the capillaries 102 can be adjusted by exchanging the capillaries 102 with capillaries 102 having different lengths. The capillary array 101 includes a sample introduction unit 104 that introduces a sample into the capillary 102, an irradiation unit 103 that irradiates the separated sample with excitation light, and a capillary head 107 that bundles a plurality of capillaries.

  The capillary 102 is a thin 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 to improve the strength. The electrophoresis path is formed by filling the inside of the capillary 102 with the electrophoresis medium. By applying a voltage to both ends of the electrophoresis path, the sample can be electrophoretically separated.

  In this embodiment, a quartz pipe whose outer surface is coated with fluorine is used so that light generated in the capillary 102 propagates through the inside. In the irradiation unit 103 slightly away from the sample introduction unit 104, the fluorine coating is removed in order to efficiently irradiate the sample in the capillary 102 with the laser beam 112. A plurality of capillaries 102 are bundled to form a capillary array 101. The number of capillaries 102 is, for example, 384, 192, 96, 16, 8, etc. If necessary, the capillary 102 may be coated with polyimide.

  The sample introduction unit 104 has the sample introduction end 105 of the capillary 102 arranged in alignment with the well of the sample container 147, and can introduce a plurality of samples held in each well into the migration path.

  In this embodiment, the sample introduction end 105 is formed by inserting the tip of the capillary 102 into the hollow electrode 106, and the sample introduction end 105 is arranged in 24 rows and 16 columns to constitute the sample introduction portion 104. At the sample introduction end 105, the tip of the capillary 102 protrudes slightly from the hollow electrode 106. The hollow electrode 106 composed of a stainless steel pipe is electrically connected to a high voltage power source 162. By immersing the sample introduction end 105 in the sample and applying a voltage, the sample can be electrophoresed and introduced into the capillary. The sample introduction method is not limited to electrophoresis, and the sample may be introduced into the electrophoresis path by pressure or dispensing.

  The irradiation unit 103 is a part that irradiates the electrophoresis medium filled in the capillary 102 with the laser light 122 that is excitation light, and can irradiate the sample component subjected to electrophoretic separation with excitation light. The phosphor labeled on the sample component emits light having a wavelength depending on the sample component.

  In the present embodiment, a plurality of capillaries 102 are divided into a plurality of groups, and a plurality of capillaries 102 are arranged on a plurality of glass substrates to form several capillary arrays. In addition, the number of sets of capillary arrays is one set, two sets, three sets, four, and the like. On each glass substrate, the capillaries 102 are arranged substantially parallel to each other and substantially parallel to the glass substrate. In addition, the removed portions of the fluorine film in each capillary 102 are arranged in a straight line. Here, the fact that the capillaries 102 are substantially parallel to each other and parallel to the glass substrate means that the degree of parallelism is within a precision error. The plurality of laser beams 122 irradiated from both sides of each glass substrate by the irradiation optical unit propagate through the polyimide film removal portions in the plurality of capillaries 102, respectively, and excite the sample in each capillary 102 at the same time.

  The capillary head 107 holds a plurality of capillaries in a bundle and can be attached to and detached from the apparatus main body. As shown in FIG. 2, the capillary head 107 of the present embodiment holds the end portions 108 of the capillaries so as to form a matrix. A part of the fluorescence generated from the sample component in the irradiation unit 103 is totally reflected on the inner surface of the capillary 102 and propagates through the capillary 102, is emitted from the terminal unit 108, and is detected by the detection optical unit. Further, the capillary head 107 can be connected to the polymer filling block 152 in a pressure-resistant and airtight manner. Then, a new electrophoresis medium can be filled into the capillary 102 from the end portion 108 by the electrophoresis medium filling unit. Note that the termination units 108 may be arranged in a matrix of 96 rows and 4 columns, 64 rows and 6 columns, or 48 rows and 8 columns as needed, or may be simply converged into one.

  The irradiation optical unit is a mechanism that irradiates the irradiation unit 103 of the capillary array 102 with laser light 122 that is excitation light. The irradiation optical unit of this embodiment includes a laser light source 112, a beam splitter A124, a mirror A125, a beam splitter B126, a mirror B127, and a condenser lens 128.

  The laser light source 112 can oscillate laser light that is excitation light for exciting sample components. The beam splitter A124 and the mirror A125 divide the laser beam 122 into two parts so that the capillary array of the irradiation unit 103 can be irradiated from both sides. The plurality of beam splitters B 126, the plurality of mirrors B 127, and the condenser lens 128 adjust the plurality of laser beams 122 and irradiate the irradiation unit 103 with the laser beams 122. Laser light 122 applied to each capillary array propagates to adjacent capillaries 102 one after another.

As the laser light source 112, a liquid laser, a gas laser, or a semiconductor laser can be used as appropriate, and an LED may be substituted if necessary. For example, an argon ion laser is a multiline that oscillates a laser beam 122 with wavelengths of 488 nm and 514.5 nm and an output of 100 mW. In addition to a multiline, a single line may be used, or two or more types of laser light sources may be combined. The wavelength of the laser beam 122 is 532 nm or 633.
nm can also be used as appropriate. Further, the laser beam 122 may be irradiated only from one side of the capillary array, or the laser beam 122 may be irradiated in a time division manner.

  The detection optical unit 131 is a mechanism for detecting light emitted from the sample component. As shown in FIG. 5, the detection optical unit 131 of this embodiment includes a first camera lens 118, an optical prism 134, a second camera lens 135, and a two-dimensional detector 136.

  The end face of the capillary head 107 having the end portion 108 is filled with the electrophoresis medium and disposed so as to face the detection window 132 made of non-fluorescent quartz glass through the polymer flow path 154. The light emitted from the end portion 108 passes through the migration medium in the polymer flow path 154, is radiated from the detection window 132, enters the first camera lens 133, and becomes a parallel light flux. The parallel light flux is wavelength-dispersed by the optical prism 134 and imaged on the two-dimensional detector 136 which is a CCD camera by the second camera lens 135. Then, the signal from the CCD camera is processed by a computer to analyze the sample.

  Instead of the optical prism 134, the wavelength dispersion means may be configured by appropriately combining diffraction gratings and filters. The two-dimensional detector 136 may be configured by appropriately combining a one-dimensional detector, a photomultiplier, a photodiode, and the like and an optical mechanism. The autosampler unit is a mechanism that transports and holds each container used for electrophoretic analysis including the sample container 147 to a predetermined position such as directly below the sample introduction unit 104. The autosampler unit of the present embodiment transports the sample container 147, the buffer container 144, the cleaning container 145, and the waste liquid container 146 by the robot arm 142 having the claws 143.

  The robot arm 142 includes a claw 143 for fixing each container thereon, and can move three-dimensionally. As a result, each container stored in a predetermined place can be transported directly below the sample introduction unit 104, held at that place for a predetermined time, and returned to the predetermined place.

The buffer container 144 is a container that holds a buffer solution that immerses the sample introduction end 105. During the electrophoresis analysis, the buffer container 144 is transported directly below the sample introduction unit 104 so that the buffer solution immerses the sample introduction end 105. Further, during the standby of the apparatus, the sample is transported in the same manner as in the electrophoresis analysis, and the sample introduction end 105 is immersed in a buffer solution to prevent the electrophoresis medium in the capillary 102 from being dried.

  The cleaning container 145 is a container that holds a cleaning liquid for cleaning the sample introduction end 105, and is transported directly below the sample introduction unit 104 after loading of the electrophoresis medium, preliminary migration, and sample introduction. By immersing the sample introduction end 105 in the cleaning liquid in the cleaning container, the sample introduction end 105 can be cleaned and contamination can be avoided.

  The waste liquid container 146 is a container for holding a used electrophoresis medium, and receives the used electrophoresis medium that is transported directly below the sample introduction unit 104 when filling the electrophoresis medium and discharged from the sample introduction end 105 when filling the electrophoresis medium. .

The sample container 147 is a container that holds a plurality of trace samples, and is transported immediately below the sample introduction unit 104 when the sample is introduced. In this embodiment, a sample container 147 is configured by placing a septa, which is a resin sheet, on a sample plate having 24 rows and 16 columns of wells capable of holding a sample of several tens of μl, and sandwiching it with a holder and a clip. ing. The sample is, for example, a solution that is fluorescently labeled so as to identify four types of nucleoside base molecules and contains a large number of nucleic acids having an appropriate length (size). The septa usually have a sealed through hole at a position corresponding to the well, and the sample introduction end 105 and the sample can be contacted at the time of sample introduction while preventing the sample in the well from evaporating. Moreover, a protective film can be stuck on the upper surface of the septa to prevent sample evaporation. Further, the holder and the clip sandwich the sample plate and the scepter therebetween, and are integrated to form the sample container 107 that can be transported by the robot arm 142.

  The electrophoresis medium filling unit shown in FIGS. 1 and 3 is a mechanism for filling the capillary 102 with a polymer as the electrophoresis medium. The electrophoresis medium filling unit of this embodiment includes a polymer filling block 152, a syringe 153, a tube 155, and an electromagnetic valve 156, and can automatically fill the capillary 102 with a new electrophoresis medium before starting analysis.

The polymer filling block 152 has a polymer flow path 154 and a detection window 132, is connected to the syringe 153 and the tube 155, and the capillary head 110 can be attached and detached. The capillary head 110 is mounted on the polymer filling block 152 while maintaining pressure and air tightness. The terminal portion 108 of the capillary head 107 is disposed so as to face the detection window 132, and a region sandwiched between the capillary head 107 and the detection window 132 forms a part of the polymer flow path 154. The polymer flow path 154 communicates with a syringe 153 filled with a migration medium and a tube 155 provided with an electromagnetic valve 156. The other end of the tube 155 is immersed in a buffer solution held in the anode buffer container 163.

  When filling the capillary 102 with the electrophoresis medium, the waste liquid container 146 is disposed immediately below the sample introduction unit 104, the electromagnetic valve 156 is closed, and the plunger of the syringe 153 is pushed. As a result, the electrophoresis medium in the syringe 153 flows into the capillary 102 from the terminal end portion 108 via the polymer channel 154. Further, the used electrophoresis medium in the capillary 102 is discharged from the sample introduction end 105 and received by the waste liquid container 146.

  The power supply unit is a mechanism that applies a voltage to the migration path composed of the migration medium in the capillary 102, and can electrophorese the sample. The power supply unit of this embodiment includes a high voltage power supply 162 that is electrically connected to the hollow electrode 106 and the anode electrode 164 and can generate a high voltage of around 15 kV.

  At the time of sample introduction, the inside of the capillary 102, the polymer channel 154, and the tube 155 is filled with the electrophoresis medium, the sample introduction end 105 is immersed in the sample held in the well of the sample container 147, and the electromagnetic valve 156 is opened. As a result, an energization path comprising the hollow electrode 106, the sample in the well, the capillary 102, the polymer flow path 154, the tube 155, the buffer solution in the anode buffer container 163, and the anode electrode 164 is formed. Then, a pulse voltage is applied to this energization path with the hollow electrode 106 as a negative potential and the anode electrode 164 as a positive potential. As a result, a negatively charged sample component, such as DNA, present in the well is introduced from the sample introduction end 105 into the migration path.

  Also, during the electrophoresis analysis, unlike the sample introduction, the sample introduction end 105 is immersed in a buffer solution held in the buffer container 144. As a result, a current path including the hollow electrode 106, the buffer solution in the buffer container 144, the capillary 102, the polymer channel 154, the tube 155, the buffer solution in the anode buffer container 163, and the anode electrode 164 is formed. Then, unlike the sample introduction, a high voltage of around 15 kV is applied. As a result, an electric field is generated in the direction from the irradiation unit 103 to the sample introduction unit 104, and the negatively charged sample component introduced into the migration path is electrophoresed in the direction of the irradiation unit 103.

  The temperature control unit is a mechanism for controlling the temperature of the migration path that affects the migration speed of the sample components. The temperature control unit in the present embodiment houses the capillary 102 in a thermostat (not shown). Then, the air maintained at a constant temperature by a temperature control mechanism such as Peltier is circulated in the thermostatic chamber by a blowing mechanism such as a fan, and the capillary 102 is maintained at a predetermined temperature.

  Hereinafter, the basic procedure of electrophoretic analysis will be described mainly with reference to FIG.

  The basic procedure of electrophoretic analysis can be broadly divided into pre-preparation 201, electrophoresis medium filling 203, preliminary electrophoresis 204, sample introduction 205, and electrophoresis analysis 206. The operator of this apparatus sets a sample and a reagent in this apparatus as a preliminary preparation before starting analysis (201). More specifically, first, the buffer container 144 and the anode buffer container 163 are filled with a buffer solution that forms part of the current path. The buffer solution is, for example, an electrolyte solution that is commercially available for electrophoresis from various companies.

  In addition, a sample to be analyzed is dispensed into the well of the sample container 147. The sample is, for example, a DNA PCR product. In addition, a cleaning solution for cleaning the sample introduction unit 104 is dispensed into the cleaning container 145. The cleaning solution is pure water, for example. Further, a separation medium for electrophoresis of the sample is injected into the syringe 153. The electrophoresis medium is, for example, a polyacrylamide separation gel commercially available for electrophoresis from various companies. Furthermore, when the deterioration of the capillary 102 is expected or when the length of the capillary 102 is changed, the capillary array 101 is replaced. Then, after the preliminary preparation is completed, the operator operates the apparatus and starts analysis (202). The electrophoresis medium filling 203 is a procedure for filling the capillary 102 with a new electrophoresis medium and forming an electrophoresis path.

  In the loading of the loading medium 203 in the present embodiment, first, the waste solution 146 is carried directly under the sample introduction unit 104 by the autosampler unit so that the used migration medium discharged from the sample introduction end 105 can be received (211). ). Then, the syringe 153 is driven to fill the capillary 102 with a new electrophoresis medium, and the used electrophoresis medium is discarded (212). Finally, the sample introduction end 105 is immersed in the cleaning solution in the cleaning container 145, and the sample introduction end 105 soiled with the electrophoresis medium is washed (213). The preliminary electrophoresis 204 is a procedure for applying a predetermined voltage to the electrophoresis medium so that the electrophoresis medium is in a state suitable for electrophoresis. In the preliminary electrophoresis 204 in the present embodiment, first, the sample introduction end 105 is immersed in the buffer solution in the buffer container 144 by the autosampler unit to form a current path (214). Then, a voltage of about several to several tens of kilovolts is applied to the electrophoresis medium for several to several tens of minutes by the power supply unit to bring the electrophoresis medium into a state suitable for electrophoresis (215). Finally, the sample introduction end 105 is immersed in the washing solution in the washing container 145, and the sample introduction end 105 soiled with the buffer solution is washed (216). The sample introduction 205 is a procedure for introducing sample components into the migration path.

  In the sample introduction 205 in the present embodiment, first, the sample introduction end 105 is immersed in the sample held in the well of the sample container 147 by the autosampler unit. As a result, an energization path is formed, and the sample component is introduced into the migration path (217). Then, a pulse voltage is applied to the energization path by the power supply unit, and the sample component is introduced into the migration path (218). Finally, the sample introduction end 105 is immersed in the cleaning solution in the cleaning container 145, and the sample introduction end 105 contaminated with the sample is washed (219). The migration analysis 206 is a procedure for separating and analyzing each sample component contained in a sample by electrophoresis. In the electrophoretic analysis 206 in the present embodiment, first, the sample introduction end 105 is immersed in the buffer solution in the buffer container 144 by the autosampler unit to form a current path (220).

  Then, a high voltage of about 15 kV is applied to the energization path by the power supply unit to generate an electric field in the migration path (221). Due to the generated electric field, each sample component in the migration path moves to the irradiation unit 103 at a speed depending on the property of each sample component. That is, the sample components are separated by the difference in moving speed. And it detects in order from the sample component which reached the irradiation part 103. For example, when the sample includes a large number of DNAs having different base lengths, a difference occurs in the moving speed depending on the base length, and the irradiation unit 103 is reached in order from the DNA having the shorter base length. If a fluorescent substance depending on the terminal base sequence is attached to each DNA, the terminal base sequence can be detected in the order of arrival at the irradiation unit 103.

When the planned data is taken, the voltage application is stopped and the electrophoresis analysis is finished (213).

  The above is a series of analysis procedures. When further analysis is performed, the analysis procedure is advanced from the loading of the migration medium 203.

  Hereinafter, the characteristic mechanism of the present embodiment will be described with reference to FIGS.

  In this embodiment, the capillary ends are arranged so as to correct the curve of the spectral line by the detection method using the spectroscopic element, and the emission spectrum of each capillary on the two-dimensional detector is mapped in a lattice pattern. With respect to the two-dimensional arrangement of the capillary ends, when the wavelength dispersion and the capillary position direction are the X-axis and only the capillary position direction is the Y-axis, the end of the capillary is arranged so as to be convex toward the short wavelength side with respect to the X-axis. .

  Further, in this embodiment, the capillary ends are arranged with high accuracy and the polymer can be filled by the polymer filling mechanism. Specifically, a spacer member is inserted in the ferrule portion at the end of the capillary array, and the position of the end portion of the capillary is fixed. In the spacer, holes having the same diameter as the capillary outer diameter are provided at intervals of the arrangement pitch. The arrangement of the end of the capillary is accurately positioned with a spacer member and fixed with an adhesive. By ensuring the alignment accuracy of the ends of each capillary, the distortion of the emission spectrum image mapped on the two-dimensional detector is eliminated. Each capillary is accurately projected in a grid on the two-dimensional detector. From the spectrally resolved images of the capillaries imaged in a grid pattern on the two-dimensional detector, information on the wavelength region necessary for analysis is acquired and processed in a lump.

  A method for arranging capillaries at the end of the capillary array will be described with reference to FIGS.

  At the end of the capillary array, a total of 48 capillaries of 6 rows in the wavelength direction and 8 rows in the capillary array direction are bundled to form a capillary array head. A spacer 501 is used for positioning each capillary. The spacer 501 is provided with the same hole as the outer diameter of the capillary in order to arrange the end of the capillary with high accuracy. The outer diameter and arrangement accuracy of the holes are secured with a tolerance of μm. For example, metal etching is used to make the hole. By metal etching, inexpensive spacers can be manufactured in large quantities with high quality.

  Due to the polymer filling mechanism in the present embodiment, a pressure of several hundred MPa is applied to the capillary end. Therefore, the structure of the capillary head is a pressure-resistant structure while ensuring the arrangement accuracy of the capillaries. In the capillary head, the capillary 503, the spacer 501, and the ferrule 502 are fixed by an adhesive 504. In this example, an epoxy adhesive (Araldite) was used. Here, each capillary detection end face and the surface of the adhesive are arranged substantially in a plane to form a detection plane. As shown in FIG. 6, the capillary detection end faces are arranged in a 6 × 8 curved lattice shape on the detection plane. For the two-dimensional arrangement at the end of the capillary, in order to correct the curvature of the spectral line by the detection method using the spectroscopic element, the chromatic dispersion and the capillary position direction are set to the X-axis and only the capillary position direction for the two-dimensional arrangement at the end of the capillary. In the case of the Y axis, the end of the capillary is arranged so as to be convex toward the short wavelength side with respect to the X axis.

  The degree of curvature of the capillary end arrangement is formulated with reference to FIG.

In a detection system using a spectroscopic element (for example, a prism), the curvature radius ρ _P of an image curved on a two-dimensional detector can be expressed by the following equation (1).

  N = n ′ / n is the refractive index of the prism with respect to air, A is the apex angle of the prism, and f ′ is the focal point of the camera lens.

(Reference: pencil of light)
N is a function of the wavelength, and is a value depending on the incident angle to the prism.

  Therefore, in order to correct this radius of curvature, an arrangement of a capillary end convexly curved toward the short wavelength side is proposed. The degree of bending of the capillary end is determined using the above formula at the center wavelength of the wavelength region to be used. For example, when the used wavelength region is 500 to 700 nm, the bending radius on the two-dimensional detector is calculated from the wavelength of 600 nm, and the bending arrangement of the capillary end is determined so as to be minimized.

  The deviation of the capillary X axis at a distance D from the capillary array center in the Y axis direction can be expressed by the following equation (2).

  When fluorescence detection at the end of the capillary is performed using the detection end, the two-dimensional detector obtains 6 emission spectra in the direction of the capillary array and 8 emission spectra in the direction of the capillary array as shown in FIG. It becomes possible to do. Since the capillary ends are arranged with high accuracy, the fluorescence wavelength components necessary for the analysis can be obtained without overlapping the fluorescence components of the capillaries.

  The acquired data processing on the two-dimensional detector is shown using FIG. In this embodiment, a 512 × 512 pixel CCD element is used. The CCD has a structure in which data is sequentially transferred to a data array buffer region in the capillary array direction or wavelength direction and processed (FIG. 9 is a schematic diagram in which data is sequentially transferred in the capillary array direction).

  The CCD element acquires the emission spectrum information of each capillary corresponding to FIG. Since the elements are arranged in a grid, if the emission spectrum image on the CCD is in a grid for each capillary, the information on the elements can be collected for each capillary as information.

  Of the elements that have acquired the spectral information of each capillary (wavelength direction 85 pixels), only the elements that have acquired the fluorescence wavelength data necessary for the analysis need only be processed in the buffer region. For example, the analysis can be performed by processing only the wavelength data of four fluorescent dyes. According to the present invention, since information about each capillary is aligned in the wavelength direction and the arrangement direction, only the pixel region necessary for the analysis needs to be performed in the wavelength direction and the arrangement direction when acquiring the spectrum image. Therefore, it is possible to speed up data processing.

  Further, since the positions of the capillaries are fixed in advance, the calibration operation of 48 capillaries and wavelengths can be easily performed when the capillaries are initially attached.

  In the present embodiment, the number of capillaries to be measured simultaneously is 48. However, in the apparatus using the present invention, a larger number of capillaries can be detected simultaneously and with high sensitivity, and capillaries in which the above-described capillary array direction is arranged in 64 rows. If an array is used, 384 capillaries can be measured simultaneously.

  The effect of the present invention will be schematically described with reference to FIG. As shown in FIG. 10A, when the capillary ends are arranged in a line, in a detector using a camera lens or a diffraction grating (prism), the fluorescence spectrum image for each capillary on the two-dimensional detector is curved. To do. In the capillary array according to the present invention, as shown in FIG. 10B, the fluorescence spectrum image for each capillary on the two-dimensional detector is arranged on a straight line. Therefore, the data processing on the detector can be speeded up.

  In the present embodiment, for the purpose of simplifying the capillary head assembling operation, the spacer with holes for capillary array in FIG. 5 is replaced with a plate-like spacer having a gentle convex surface (FIG. 11A), a spacer with a V groove (FIG. 11 (b)). In FIG. 11A, a plurality of plate-like spacers are arranged at equal intervals, and a plurality of plate-like spacers having convex surfaces are arranged so as to be substantially perpendicular to them. Capillaries are arranged and fixed in each region partitioned by the spacers. In FIG. 11B, a plurality of V-grooves having a V-shaped cross section are provided in the spacer, and each capillary is fitted into the V-groove, and a plurality of the spacers are stacked. The V-groove can hold the capillary with high accuracy, but other than this, a concave groove, a round groove, or the like can be used as appropriate. These spacers can be mass-produced with high accuracy by etching a silicon substrate. According to this embodiment, it is possible to perform a manufacturing method in which capillaries are simply arranged on plate-like spacers and fixed as they are with a ferrule member and an adhesive.

  In this embodiment, an arrangement method for correcting distortion of a spectral image on a two-dimensional detector due to aberration due to characteristics of a lens used in a detection unit will be described. Only the capillary end arrangement according to the first embodiment may cause distortion as shown in FIGS. 12A and 12B due to lens characteristics. Therefore, in order to correct the distortion, in the case of the pincushion type as shown in FIG. 12A, as shown in FIG. On the contrary, in the case of the barrel type as shown in FIG. 12B, as shown in FIG. 12D, it is adjusted to a pincushion type and the capillary ends are arranged.

  Although the embodiments have been described above, the present invention is not limited thereto, and it is understood by those skilled in the art that various modifications can be made within the scope of the invention described in the claims. . For example, it is naturally possible to appropriately combine the embodiments.

The schematic perspective view of an electrophoresis apparatus. The schematic perspective view of a capillary head. The schematic sectional drawing of the polymer filling block vicinity. Schematic flow chart of analysis procedure. FIG. 2 is a schematic view of a capillary end according to the first embodiment. 2 is a schematic diagram of a two-dimensional array according to Example 1. FIG. FIG. 3 is a diagram of formulation of a two-dimensional array according to the first embodiment. 2 is an image on a two-dimensional detector according to Example 1. FIG. FIG. 6 is a diagram of processing of the two-dimensional detector according to the first embodiment. The figure which shows the spectrum curve prevention effect by Example 1. FIG. 2 is a schematic diagram of a two-dimensional array according to Example 2. FIG. 4 is an image on a two-dimensional detector according to Example 3. FIG.

Explanation of symbols

DESCRIPTION OF SYMBOLS 101 ... Capillary array, 102,503 ... Capillary, 103 ... Irradiation part, 104 ... Sample introduction part, 105 ... Sample introduction end, 106 ... Hollow electrode, 107 ... Capillary head, 108 ... Terminal part, 121 ... Irradiation optical unit, 122 DESCRIPTION OF SYMBOLS ... Laser beam, 123 ... Laser light source, 124 ... Beam splitter A, 125 ... Mirror A, 126 ... Beam splitter B, 127 ... Mirror B, 131 ... Detection optical unit, 132 ... Detection window, 133 ... First camera lens, 134 ... optical prism, 135 ... second camera lens, 136 ... two-dimensional detector, 141 ... autosampler unit, 142 ... robot arm, 143 ... claw, 144 ... buffer container, 145 ... cleaning container, 146 ... waste liquid container, 147 ... Sample container 151... Migration medium filling unit 152. Polymer filling block 1 DESCRIPTION OF SYMBOLS 3 ... Syringe, 154 ... Polymer flow path, 155 ... Tube, 156 ... Solenoid valve, 161 ... Power supply unit, 162 ... High voltage power supply, 163 ... Anode buffer container, 164 ... Anode electrode, 501 ... Spacer, 502 ... Ferrule, 504 …adhesive.

Claims (9)

A plurality of capillaries capable of being filled with an electrophoresis medium;
A power source for electrophoresis of the sample in the electrophoresis medium;
An excitation optical system for irradiating the separated sample with excitation light;
A light receiving optical system including a lens and a two-dimensional detector for detecting light emitted from a plurality of capillary ends arranged in a substantially matrix shape;
An electrophoresis apparatus comprising:
An electrophoresis apparatus in which at least three capillary ends are arranged in a curved shape in a direction perpendicular to a wavelength dispersion direction . Oite the electrophoresis apparatus according to claim 1, row or a plurality of capillary end corresponding to the column, the electrophoresis apparatus characterized by being arranged in a convex curved shape. The electrophoresis apparatus according to claim 1, wherein a plurality of capillary ends arranged in a substantially matrix form are arranged so as to gather at a central portion . 2. The electrophoresis apparatus according to claim 1, wherein a plurality of capillary ends arranged in a substantially matrix form are arranged so as to swell outward .   2. The electrophoretic device according to claim 1, wherein in the two-dimensional detector, light emission is substantially linear from a plurality of capillary ends corresponding to rows or columns. 2. The electrophoretic device according to claim 1, wherein the light receiving optical system includes wavelength dispersion means.   2. The electrophoretic device according to claim 1, wherein the light receiving optical system includes a condenser lens, a diffraction grating, and an imaging lens.   The electrophoresis apparatus according to claim 1, further comprising a spacer for fixing a plurality of capillary ends. 2. The electrophoresis apparatus according to claim 1, further comprising means for correcting aberration caused by the lens.

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