Capillary electrophoresis device

Abstract

In order to make a light detection optical system less susceptible to the external environment, this capillary electrophoresis device is provided with a light source, a first irradiation fiber that guides light from the light source, a detector that detects light from a capillary, and a first detection fiber that guides light to the detector. A capillary cartridge having a capillary, the second irradiation optical fiber, and the second detection optical fiber is attached to the capillary electrophoresis device by connecting the first irradiation optical fiber and the second irradiation optical fiber and connecting the first detection optical fiber and the second detection optical fiber. As shown in Figure 4, a capillary electrophoresis device is proposed, in the capillary cartridge, the second illumination fiber and the second detection fiber are fixed such that their optical axes intersect in the lumen of the capillary.

Description

Technical Field

[0001] The present invention relates to an apparatus for optically detecting an analyte in a capillary, particularly in a capillary electrophoresis apparatus. Background Art

[0002] In an analysis using capillary electrophoresis, an analyte sample is injected into a capillary filled with a separation medium, a voltage is applied across both ends, and separation is performed based on the difference in the migration rates of the analytes. There are multiple means for measuring the separated sample, including a method of detecting fluorescence emitted from the sample and a method of detecting light absorption of the sample.

[0003] For example, there is a method of electrophoresing DNA labeled with a fluorescent dye in a capillary filled with a polymer and separating it by each chain length. An excitation light is irradiated onto a detection site provided on the capillary, and the generated fluorescence is detected. The DNA molecules in the sample move in the capillary and pass through the detection site at different times according to their chain lengths. As a result, the chain length distribution of the DNA molecules in the sample is obtained as a fluorescence intensity waveform.

[0004] In a capillary electrophoresis apparatus, the inner diameter of the capillary in which sample separation is performed is generally about several tens of micrometers. In order to improve the detection sensitivity, it is preferable to irradiate the inner diameter of the capillary through which the sample passes with fluorescence excitation light with as little loss as possible. Therefore, it is preferable to condense the excitation light to the same degree as or smaller than the inner diameter of the capillary and irradiate it onto the inner diameter of the capillary. At this time, it is necessary to adjust the positions of the capillary and the condensed excitation light with a positional accuracy equal to or higher than the size of the inner diameter of the capillary. In particular, in a configuration where a plurality of capillaries are arranged in a row and all the capillaries are excited by irradiating excitation light from the side, the excitation light needs to pass through the inner diameters of all the capillaries with as little loss as possible, so the requirement for positional accuracy becomes stricter, and the positional error needs to be set to about 10 μm or less.

[0005] In addition, when there are multiple types of fluorescent dyes used to label the measurement object, the generated fluorescence is introduced into a spectroscopic optical system, and after wavelength separation, it is measured by an imaging element. As an example, spectroscopy is performed using a grating. However, in this case, when the position of the light-emitting point in the capillary changes in the wavelength separation direction, a wavelength shift of the fluorescence is observed on the detector. Since the type of fluorescent dye emitted is determined based on the spectral shape of the measured light, when the fluorescence wavelength shifts, the discrimination accuracy of the fluorescent dye decreases. A decrease in the pigment discrimination accuracy may lead to false detections such as false detection of DNA strands that do not actually exist in DNA analysis, which is not preferable.

[0006] As described above, the excitation light irradiation optical system, the capillary, and the detection optical system need to be adjusted with high precision so that their positional relationship is within an error range of several tens of micrometers or less. In addition, it is not desirable for the detection performance of the device to change, and it is also desirable to maintain the positional relationship during long-term use of the device and when the device is moved. On the other hand, in capillary electrophoresis, the capillary is a consumable item and deteriorates after a certain number of measurements, so it needs to be replaced. Therefore, even when the capillary is replaced, the above-described positional accuracy needs to be maintained. From the viewpoint of user convenience, it is desired to be able to easily perform the capillary replacement.

[0007] Prior Art Documents

[0008] Patent Documents

[0009] Patent Document 1: Japanese Patent Laid-Open No. 8-304339

[0010] Patent Document 2: Japanese Patent Application Laid-Open No. 2004-532384

[0011] Patent Document 3: US2021 / 0003530

[0012] Non-Patent Documents

[0013] Non-Patent Document 1: H. Zhai et al., “A simple and compact fluorescence detection system for capillary electrophoresis and its application to food analysis,” Electrophoresis, 36, 2509 (2015). Summary of the Invention

[0014] Problems to be Solved by the Invention

[0015] Generally, a capillary used in electrophoresis, an optical system for irradiating light to the capillary, and an optical system for detecting light are installed and fixed inside an electrophoresis apparatus. However, due to vibrations and impacts transmitted from the outside when the apparatus is moved, etc., the positions of the respective components and the optical elements inside them may change. In addition, even when no vibrations or impacts are applied, due to external environments, especially the expansion and contraction of the respective components caused by temperature changes, changes in the positions of the optical elements may also occur.

[0016] The position of the capillary may also change due to the replacement of the capillary. The position of the capillary may change compared to before the replacement due to the tolerance of the outer diameter of the capillary, the error of the fixed position caused by the capillary fixing mechanism, etc. In the case of a plurality of capillaries, the capillary array is often provided to the user in a state where the capillary is fixed to a fixing member, but the position of the capillary array may change due to the error of the assembly of the fixing member and the main body.

[0017] A method for reducing the occurrence and influence of the above-mentioned positional error by assembling an optical fiber optical system on an electrophoresis flow path is proposed. For example, the following method is disclosed in Patent Document 1: by adopting a method in which an optical fiber for excitation and detection is set on a flow path chip, and a capillary as a consumable is installed on the flow path chip, even if the capillary is replaced, the relative position relationship of the detection optical system does not change. However, there is the following problem in this method. In the case where a gap is generated between the flow paths at the junction of the capillary and the flow path chip, and the position of the central axis of the two is offset, the separation performance of electrophoresis is adversely affected. Therefore, it is preferred to detect the sample after electrophoresis at a detection point set on a part of the capillary. In addition, in the method of Patent Document 1, when multiple capillaries are carried, it is necessary to carry multiple disclosed structures, the device is large-scale, and the excitation light must be branched according to the number of detection optical fibers, so there is a problem of reduced power of the excitation light and reduced detection performance.

[0018] Patent document 2 describes a method for reducing the alignment accuracy required when replacing a capillary by providing a box in which optical components such as optical fibers and lenses are assembled to the capillary. In the structure disclosed in patent document 2, a detection fiber array including a light source such as an LED and a lens is connected to a capillary box with an irradiation optical fiber built in. In this structure, the relative positional relationship between the capillary and the irradiation optical fiber is difficult to change. However, the position adjustment accuracy of the capillary and the detection fiber array, the light source and the irradiation optical fiber when replacing the box depends on the accuracy of the detachable mechanical fixing mechanism. In addition, if the capillary box is considered to be a consumable, its mechanical fixing mechanism needs to be cheap. In addition, the mechanical fixing mechanism is robust to vibrations and shocks from the outside, and the positional relationship of each component needs to be kept constant at all times. In patent document 2, there is no disclosed fixing mechanism that meets the above requirements.

[0019] Non-Patent Document 1 describes a method of performing detection by fixing a capillary and an irradiation detection optical fiber to a grooved flat plate formed of polydimethylsiloxane. In this method, since the capillary and the optical element are fixed to the same base, it is difficult for the position of the optical element to shift due to vibration or shock. However, a method for replacing the capillary is not described, nor is a method for adjusting the relative positional relationship between the capillary and the optical system during capillary replacement. In addition, when the number of capillaries is plural, the same problems as those in Patent Document 1 may occur.

[0020] In addition, in capillary electrophoresis, it is necessary to uniformly adjust the temperature of the capillary. As an example, when analyzing DNA by capillary electrophoresis, in order to perform electrophoresis in a state where DNA is denatured, the temperature of the capillary is maintained at about 60°C. In order to maintain the high separation ability of electrophoresis and, in addition, to obtain the same measurement result for the same sample each time, the entire capillary needs to have a uniform temperature and small temperature fluctuations. This temperature adjustment mechanism also needs to balance temperature control performance and ease of capillary replacement.

[0021] Regarding the temperature adjustment of the capillary, Patent Document 1 and Patent Document 2 each describe a method for adjusting the temperature of the capillary, but do not describe that the disclosed structure is particularly advantageous for temperature adjustment. Non-Patent Document 1 does not describe the temperature adjustment of the capillary.

[0022] Means for Solving the Problem

[0023] As an example of the capillary electrophoresis apparatus of the present invention, the capillary electrophoresis apparatus includes a light source, a first irradiation optical fiber that guides light from the light source, a detector that detects light from the capillary, and a first detection optical fiber that guides light to the detector.

[0024] By connecting the first irradiation optical fiber and the second irradiation optical fiber, and connecting the first detection optical fiber and the second detection optical fiber, a capillary cassette having a capillary, the second irradiation optical fiber, and the second detection optical fiber is attached to the capillary electrophoresis apparatus.

[0025] In the capillary cassette, the optical axes of the second irradiation optical fiber and the second detection optical fiber are fixed so as to intersect inside the lumen of the capillary.

[0026] Advantageous Effects of the Invention

[0027] According to the capillary electrophoresis apparatus of the present invention, it is possible to make the optical detection optical system in the capillary electrophoresis apparatus less susceptible to changes in the external environment, shock, and vibration.

[0028] In addition, since the optical systems of the main body and the capillary cassette can be connected by connecting the optical fibers, the capillary cassette can be easily attached and detached.

[0029] In the structure of the present disclosure, it is possible to configure to isolate the detection window of the capillary from the external environment. Therefore, it is possible to adjust the temperature of the entire capillary including the measurement window with good precision.

[0030] Moreover, the structure of the present invention can also be extended when the number of capillaries is plural. By adopting a structure in which excitation light is irradiated from the side of the capillary array, even when the number of capillaries is large, a reduction in the excitation light power caused by the division of the excitation light can be prevented. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 is a structural diagram of the electrophoresis apparatus 100 according to Embodiment 1 of the present disclosure.

[0032] Figure 2 is a diagram showing the measurement process of the electrophoresis apparatus 100 according to Embodiment 1 of the present disclosure.

[0033] Figure 3 is a fixing method of the capillary 111 and the cassette side irradiation optical fiber 114 in the detection portion 116.

[0034] Figure 4 is a fixing method of the cassette side detection optical fiber 115 in the detection portion 116.

[0035] Figure 5 is a fixing method of the capillary 111, the cassette side irradiation optical fiber 114, and the side detection optical fiber 115 in the detection portion 116.

[0036] Figure 6 is a schematic diagram showing the connection portion of the capillary cassette 110.

[0037] Figure 7 is a schematic diagram showing a temperature adjustment method of the capillary 111.

[0038] Figure 8 is a structural diagram of the electrophoresis apparatus 800 according to Embodiment 2 of the present disclosure.

[0039] Figure 9 is a structural diagram of the electrophoresis apparatus 800 and the capillary cassette 801 according to Embodiment 2 of the present disclosure.

[0040] Figure 10 is a schematic diagram showing a fixing method of the detection portion 116.

[0041] Figure 11 is a diagram showing the structure of the detection portion 116 according to Embodiment 3 of the present disclosure.

[0042] Figure 12 is a diagram showing the arrangement of optical fibers when fluorescence measurement and absorbance measurement are taken into consideration.

[0043] Figure 13 This is a diagram showing the structure of the fixed substrate 1301 according to Embodiment 4 of the present disclosure.

[0044] Figure 14 This is a diagram showing the arrangement of the capillary 1401, the irradiation optical fiber 1402, and the detection optical fiber 1403 with respect to the fixed substrate 1301.

[0045] Figure 15 This is a diagram showing the cause of crosstalk between capillaries.

[0046] Figure 16 This is a diagram showing the effect of the fixed substrate 1301 in reducing crosstalk between capillaries. Detailed Embodiment

[0047] <Embodiment 1>

[0048] Figure 1 This is a structural diagram of the electrophoresis apparatus 100 according to Embodiment 1 of the present disclosure. This embodiment shows an example when fluorescence detection is used as the detection method. As the sample to be measured, DNA is considered, but the sample is not limited thereto. The electrophoresis apparatus 100 includes an irradiation optical system 101 that generates excitation light for fluorescence measurement, and a detection optical system 102 that detects fluorescence. In addition, as a device for performing electrophoresis, it includes: a high-voltage power supply 103; a polymer container 104 that holds a polymer as a separation medium; a pump unit 105 that fills the polymer; a temperature adjustment device 106 that adjusts the temperature of the capillary; a buffer solution container 107 that is electrically connected to both ends of the capillary and holds a buffer solution that applies the voltage of the high-voltage power supply to the capillary; and an automatic sampling unit 108. These respective constituent elements are controlled by a control device 109. A capillary cartridge 110, which is a consumable, is connected to the electrophoresis apparatus 100. The capillary cartridge has a capillary 111 inside. The high-voltage power supply 103 applies a voltage to both ends of the capillary 111 via the buffer solution.

[0049] The irradiation optical system 101 inside the electrophoresis apparatus 100 includes a main body side irradiation optical fiber 112 for guiding the generated excitation light, and the detection optical system 102 includes a main body side detection optical fiber 113 for guiding fluorescence. On the other hand, the capillary cartridge 110 includes a cartridge side irradiation optical fiber 114 for guiding excitation light to the capillary, and a cartridge side detection optical fiber 115 for guiding the fluorescence generated inside the capillary. The capillary 111, the cartridge side irradiation optical fiber 114, and the cartridge side detection optical fiber 115 are adjusted in relative position at the detection site 116 and fixed. The main body side irradiation optical fiber 112 and the cartridge side irradiation optical fiber 114, and the main body side detection optical fiber 113 and the cartridge side detection optical fiber 115 are connected by fiber optic connectors 117.

[0050] The pump unit 105 is composed of a flow path block 118, a syringe 119, a check valve 120, and a valve 121. The check valve 120 is arranged such that the fluid only flows in the direction from the polymer container 104 to the flow path block 118. The automatic sampling unit 108 is composed of a buffer tray 122, a washing water tray 123, a waste liquid tray 124, a sample tray 125, and a stage 126 that controls their positions. It is configured to dispose an electrode 127 near the sample injection end of the capillary 111, and when the buffer tray 122 and the sample tray 125 are disposed near the injection end, electrically connect the end of the capillary 111 to the high-voltage power supply 103.

[0051] Hereinafter, the mechanism of fluorescence detection by the electrophoresis apparatus 100 of the present disclosure will be described. The excitation light generated in the irradiation optical system 101 is introduced into the main body side irradiation optical fiber 112, and is introduced into the cartridge side irradiation optical fiber 114 through the optical fiber connector 117. The excitation light reaches the detection part 116 through the cartridge side irradiation optical fiber 114, and irradiates the inner diameter of the capillary 111. The fluorescence generated at the excitation light irradiation part of the capillary 111 is condensed by the cartridge side detection optical fiber 115, and is transferred to the main body side detection optical fiber 113 by the optical fiber connector 117. Thereafter, the fluorescence is detected in the detection optical system 102 and converted into an electrical signal. This signal is collected by the control device 109.

[0052] Hereinafter, the mechanism of sample analysis by electrophoresis using the electrophoresis apparatus 100 of the present disclosure will be described. As an example, the electrophoresis analysis is carried out in the order of polymer injection into the capillary, preliminary electrophoresis, sample injection, and electrophoresis of the sample. The operation steps of the apparatus in the electrophoresis analysis are as Figure 2 shown.

[0053] When the measurement is started (S201), first, the capillary 111 is filled with the polymer by the pump unit 105. First, the waste liquid tray 124 is disposed at the end of the capillary 111 (S202), and the valve 121 is closed (S203). With the valve 121 closed, the syringe 119 is made negative pressure, and the polymer is filled into the syringe 119 from the polymer container 104 (S204). In addition, the inner diameter of the capillary 111 is on the order of several tens of micrometers, which is sufficiently thin compared to the flow path diameter of the flow path block 118 and has a large resistance, so the buffer solution does not flow from the capillary 111 to the syringe 119. Then, the syringe 119 is pressurized to fill the capillary 111 with the polymer (S205). Thereafter, the valve 121 is opened, and the end of the capillary 111 is electrically connected to the high-voltage power supply 103 (S206).

[0054] After filling the capillary with a polymer, a preliminary electrophoresis is performed. The washing water tray 123 is moved to the sample introduction end of the capillary 111, and the front end is washed (S207). Next, the buffer tray 122 is set at the sample introduction end of the capillary 111 (S208), and a high voltage is applied for several minutes by the high-voltage power supply 103 to perform a preliminary electrophoresis (S209). By the preliminary electrophoresis before injecting the sample, the impurity ions of the polymer filled in the capillary are removed.

[0055] After the preliminary electrophoresis, a sample is injected into the capillary 111. First, the washing water tray 123 is moved to the sample introduction end of the capillary 111, and the front end is washed (S210). Then, the sample tray 125 is set at the sample introduction end of the capillary 111 (S211), and a short-time voltage of several seconds is applied to both ends of the capillary 111 by the high-voltage power supply, thereby electrically injecting the sample into the capillary 111 (S212). By this step, a state is achieved in which the sample is injected only into a minute area at the end of the capillary 111. Next, the washing water tray 123 is moved again to the sample introduction end of the capillary 111, and the front end is washed to remove the remaining sample adhering to the outer wall of the capillary 111 (S213). After that, the buffer tray 122 is set at the sample introduction end of the capillary 111 (S214).

[0056] After injecting the sample, electrophoresis separation of the injected sample is performed. A voltage is applied to both ends of the capillary 111 by the high-voltage power supply 103 to make the injected sample migrate (S215). During electrophoresis, the temperature control device 106 maintains the capillary 111 at a constant temperature. A phosphor is given to the sample, and fluorescence measurement is performed by the above method when passing through the detection part 116. The moving speeds of the respective components in the sample differ according to their charge amounts and molecular sizes, and there are differences in the times until reaching the detection part 116. Therefore, the time waveform of the fluorescence signal obtained by performing fluorescence measurement at the detection part 116 gives information on the components of each sample. After the signal waveform has been acquired, the voltage application is ended and the measurement is ended (S216).

[0057] Figure 3 、 Figure 4 Details of a structural example of the detection part 116 are shown. As an example, the detection part 116 can be formed by fixing the capillary 111, the cassette-side irradiation optical fiber 114, and the cassette-side detection optical fiber 115 on a substrate formed with a V-groove. Figure 3 (a) of shows the structure of the fixing substrate 301 for fixing the capillary 111 and the cassette-side irradiation optical fiber 114. The fixing substrate 301 includes a capillary fixing groove 302 and an optical fiber fixing groove 303.

[0058] Figure 3Figure (b) is an enlarged view near the intersection of the capillary fixing groove 302 and the optical fiber fixing groove 303. The optical fiber fixing groove 303 has a lens fixing groove 304 immediately before the intersection with the capillary fixing groove 302. In addition, a through-hole 305 is provided at the intersection of the optical fiber fixing groove 303 and the capillary fixing groove 302. The through-hole 305 is provided so that the laser light emitted from the optical fiber 114 irradiated from the cassette side fixed in the optical fiber fixing groove 303 is not blocked by the wall surface of the capillary fixing groove 302. In this example, the through-hole 305 penetrates the substrate, but it may also be a recess that does not penetrate the substrate. A substrate having the shape of Figure 3 Figure (a) can be formed, for example, by anisotropic etching of silicon.

[0059] Figure 3 Figure (c) shows a diagram when the capillary 111, the cassette-side irradiation optical fiber 114, and the ball lens 306 are provided on the fixed substrate 301. The capillary 111 is generally coated with a covering such as polyimide, and at the periphery of the through-hole 305, this coating becomes an obstacle to optical measurement and is removed. The excitation light guided by the cassette-side irradiation optical fiber 114 is collimated by the ball lens 306. The collimated excitation light passes through the four capillaries 111 and excites the phosphor inside the capillaries 111. In the case of adopting the Figure 3 optical structure of Figure (c), it is known that the outer diameter, inner diameter, and interval of the capillary 111 are set so that the excitation light propagates sequentially in each capillary through the lens effect of the capillary.

[0060] Generally, the outer diameters of the capillary 111, the cassette-side irradiation optical fiber 114, and the ball lens 306 are different, but by adjusting the depths of the V-grooves that fix them respectively, the central axes of the respective components are made to coincide with the substrate surface, and thus the axis alignment of the respective components can be performed. In addition, here, the ball lens 306 is used for collimating the excitation light emitted from the cassette-side irradiation optical fiber 114, but collimation may also be performed by other methods such as a GRIN lens or a lens optical fiber. In addition, in the case where the number of capillaries is one, there is also an option not to perform collimation.

[0061] Figure 4 Shows a method of further installing the cassette-side detection optical fiber 115 on the fixed substrate 301 on which the capillary 111 and the cassette-side irradiation optical fiber 114 are fixed as shown in Figure 3 . The pressing substrate 401 having a structure similar to that of the fixed substrate 301 is mounted on the fixed substrate 301 on which the respective components are provided ( Figure 4 Figure (a)). The pressing substrate 401 has the same capillary fixing groove 302 and optical fiber fixing groove 303 as the fixed substrate 301. On the other hand, it is different in structure from the fixed substrate 301 in that a detection optical fiber array mounting hole 402 is provided. The fixed substrate 301 and the pressing substrate 401 are fixed in such a way that the grooves face each other ( Figure 4 Figure (b)).Figure 4 (c)).

[0062] As an example, when the capillary 111 and the box-side irradiation optical fiber 114 are provided in the same structure as the fixed substrate 301, the capillary fixing groove 302 and the optical fiber fixing groove 303 of the pressing substrate 401 are grooves whose central axes are located on the surface of the substrate. In this structure, the capillary 111 and the box-side irradiation optical fiber 114 are fixed by being clamped by the fixed substrate 301 and the pressing substrate 401. Figure 4 In the example, the ball lens 306 is only in contact with the fixed substrate 301 and therefore needs to be fixed using an adhesive or the like. However, the structure may also be as follows: the irradiating optical fiber 114 is bonded and fixed to the box side using a transparent adhesive in advance, the distance between the ball lens 306 and the capillary 111 is increased, and the ball lens 306 is set at a position clamped by the substrate.

[0063] As an example, the box-side detection optical fiber 115 is fixed in a V-groove formed in another substrate to form a detection optical fiber array 403. The detection optical fiber array 403 is fixed in a state of being inserted into the detection optical fiber array mounting hole 402 ( Figure 4 (d)). For example, the array fixing member 404 may be fixed to the pressing substrate 401 by using an adhesive or the like, and the detection optical fiber array 403 may be fixed to the array fixing member 304 by using an adhesive or the like.

[0064] The optical fiber array 403 and the capillary 111 can be aligned by a plurality of means. For example, the size of the optical fiber array 403 can be adjusted to be consistent with the size of the optical fiber array mounting hole 402, and when the optical fiber array 403 is embedded, it can be adjusted to be fixed in a form in which the detection optical fiber 115 on the box side faces the excitation light irradiation part on the capillary 111. Alternatively, the detection optical fiber array mounting hole 402 can be a hole of the same number of capillaries opened at the position of the central axis of each capillary ( Figure 4 (e)), alignment is performed by inserting the box-side detection optical fiber 115 into the hole.

[0065] The optical fiber array 403 may be aligned using any observation means. For example, the capillary 111 and the optical fiber array 403 may be observed by a camera through the through hole 305, and the optical fiber array 403 may be fixed in a state where the positions of the two are aligned. Alternatively, water, an aqueous solution of a fluorescent dye, etc. may be injected into the capillary 111, and the Raman signal, the fluorescent signal, etc. output from the detection optical fiber 115 on the box side may be monitored while irradiating the excitation light. The position of the optical fiber array 403 may be adjusted and fixed in such a way that these signals are maximized.

[0066] exist Figure 3 , Figure 4In the figure, an example in which the number of capillary tubes is 4 is shown, but any number of capillary tubes can be used. When the number of capillary tubes is large, in order to equalize the power of the excitation light irradiated to each capillary tube, two box-side detection optical fibers 115 can be provided, and the excitation light can be irradiated from both side surfaces of the capillary array.

[0067] When the number of capillary tubes is 2 or less, the capillary tube 111, the box-side irradiation optical fiber 114, and the box-side detection optical fiber 115 can be provided on the same substrate. Figure 5 (a) of shows a structural example of the fixed substrate 501 when the number of capillary tubes is 2. In this example, the fixed substrate 501 has a capillary tube fixing groove 502, an irradiation optical fiber fixing groove 503, a ball lens fixing hole 504, and a detection optical fiber fixing groove 505. These grooves are formed, for example, by dry-etching a silicon substrate to excavate grooves having a quadrilateral cross-section. When the groove is quadrilateral, the depth or width of the groove is adjusted so that the central axes of the capillary tube 111, the box-side irradiation optical fiber 114, the ball lens 306, and the box-side detection optical fiber 115 are in the same plane.

[0068] Figure 5 (b) of is a view showing the capillary tube 111, the box-side irradiation optical fiber 114, the ball lens 306, and the box-side detection optical fiber 115 provided on the fixed substrate 501. After the respective components are aligned by the grooves, they are fixed by an adhesive or the like. The capillary tube 111 is the same as the example of Figure 3 and the covering is removed at the detection position.

[0069] The structure of the detection part 116 is not limited to the above structure, and other structures can also be adopted as long as the relative positions of the capillary tube 111, the box-side irradiation optical fiber 114, and the box-side detection optical fiber 115 can be fixed. It is not necessarily required to perform alignment through the structure and grooves on the substrate. For example, it can also be a method of adjusting the positions of the capillary tube or the optical fiber by a jig or the like on the substrate and fixing them with an adhesive.

[0070] In addition, the box-side irradiation optical fiber 114 and the box-side detection optical fiber 115 do not necessarily need to be arranged such that their central axes face the detection point on the capillary tube 111. For the box-side irradiation optical fiber 114 and the box-side detection optical fiber 115 whose central axes do not face the detection point on the capillary tube 111, they can be configured such that the light emitted from the box-side irradiation optical fiber 114 is guided to the detection point on the capillary tube 111 by a reflecting mirror provided on the fixed substrate 501, and the fluorescence emitted from the detection point on the capillary tube 111 is incident on the box-side detection optical fiber 115.

[0071] The requirements related to the above-described optical fiber configuration are the same even when there are components other than the mirror on the fixed substrate 501. Assuming light rays emitted from the end face of the optical fiber along the central axis direction of the optical fiber, the optical elements on the fixed substrate 501 bring optical effects such as reflection, refraction, and diffraction to the light rays. At this time, the trajectory of the light rays is defined as the optical axis of the optical fiber. In order to detect the fluorescence emitted from the substance in the capillary, it is sufficient that the optical axes of the cartridge-side irradiation optical fiber 114 and the cartridge-side detection optical fiber 115 intersect in the inner cavity of the capillary 111. In addition, the above optical axes do not need to strictly intersect in the inner cavity of the capillary 111, and an error is allowed as long as it is within the range where the fluorescence can enter the cartridge-side detection optical fiber 115.

[0072] Figure 6 Describe the connection when the capillary cartridge 110 is set on the electrophoresis apparatus 100. The capillary array needs to be connected to the pump unit 105 in order to inject polymers into the interior and, in addition, to be electrically connected to the electrodes in the buffer tank 107. As an example, the capillaries 111 converge into a bundle at the connection part and are connected to the pump unit 105 using an assembly part 601 or the like.

[0073] When the electrode 127 is built in the capillary cartridge 110, the high-voltage power supply 103 and the electrode 127 are connected through the electrical connector 602. In addition, the temperature adjustment device 106 is connected via the temperature adjustment connector 603. When temperature adjustment is performed using a heater or the like built in the capillary cartridge 110, the temperature adjustment connector 603 is an electrical connector. When temperature adjustment is performed by flowing a fluid such as air, the temperature adjustment connector 603 is a connector for the flow path.

[0074] As described above, the main body-side irradiation optical fiber 112 and the cartridge-side irradiation optical fiber 114, and the main body-side detection optical fiber 113 and the cartridge-side detection optical fiber 115 are connected through the optical fiber connector 117. As the optical fiber connector 117, generally popular SC connectors, FC connectors, LC connectors, etc. can be used. In the case where the number of connected optical fibers is large, a multi-core connector such as an MPO connector can also be used. Or, it can also be a specially designed optical fiber connector.

[0075] It is configured to connect the optical systems of the electrophoresis apparatus 100 and the capillary cartridge 110 by connecting the main body-side irradiation optical fiber 112 and the cartridge-side irradiation optical fiber 114, and the main body-side detection optical fiber 113 and the cartridge-side detection optical fiber 115, thereby achieving both the easy replaceability of the capillary cartridge 110 and the resistance to vibration and external environment changes. For the optical system, the user can install and remove the optical fiber cartridge 110 only by attaching and detaching the optical fiber connector 117. In addition, since the transmission of light is through the optical fiber, it becomes robust against the influence of vibration and external environment changes.

[0076] For example, when the capillary 10 cm away from the light source is irradiated with excitation light with a position accuracy of ±10 μm by beam propagation through free space, the angle variation of the beam is only allowed to be less than ±0.01. In order to achieve this accuracy and stability, it is necessary to form the holding structure of the optical system with a strong and low thermal expansion material that will not be deformed by vibration or impact, but as a result, the device becomes large and heavy. On the other hand, if the transmission is carried out through optical fiber, it is sufficient to maintain high rigidity of the parts from the light source to the optical fiber and from the optical fiber to the capillary. In addition, the distance between the two can be set to less than a few millimeters, so the tolerance for the beam angle variation caused by the deformation of the holding structure is also increased.

[0077] The above-mentioned effects generally relate to fiber optic systems, and in particular, in the structure of the present invention, by fixing the capillary 111, the box-side irradiation optical fiber 114, and the box-side detection optical fiber 115 on the substrate, and separately providing a connection point with the outside, it is easy to take into account both the ease of loading and unloading and the fixing accuracy in the connection between the electrophoresis device 100 and the capillary box 110. In the case of a form in which the optical fiber array is loaded and unloaded from the capillary array as in Patent Document 2, the fixing structure of the optical fiber array must take into account both the ease of loading and unloading and the fixing accuracy, and the loading and unloading mechanism may become complicated and costly. On the other hand, in the structure of the present disclosure, the ease of loading and unloading and the fixing accuracy for external connection are ensured by using an optical fiber connector as a generally popular component, and it is sufficient for the capillary and the optical fiber to meet the fixing accuracy requirements, so the complexity and high cost of the loading and unloading mechanism can be avoided.

[0078] Figure 7 A method of adjusting the temperature of the capillary 111 in the capillary cartridge 110 of the present disclosure is described. Figure 7 (a) is an example in which the capillary 111 is temperature-controlled by a heating element such as a sheet heater 701. The capillary 111 and the detection portion 116 are arranged to be in contact with the sheet heater 701. The sheet heater 701 receives power supply from the electrophoresis device 100 via a temperature control connector 603. A temperature sensor for feedback control may also be provided on the sheet heater 701.

[0079] Figure 7 (b) is an example of temperature regulation of the capillary 111 by supplying a fluid such as temperature-regulated air or an inert liquid to the capillary box. In this example, the temperature regulation connector 603 is composed of a fluid supply port 702 and a fluid discharge port 703. The temperature-regulated fluid is supplied from the fluid supply port 702. A partition wall 704 is provided inside the box, and the fluid flows inside without stagnation, thereby regulating the temperature of the capillary 111. Afterwards, the fluid returns to the electrophoresis device 100 from the fluid discharge port 703.

[0080] In the case where a liquid is used as the fluid for temperature adjustment, the refractive index of the liquid is different from that of air. Therefore, when the liquid invades the detection part 116, the optical adjustment state may change. In this case, it can be configured to seal the detection part 116 so that the liquid does not invade the inside, or it can be configured that the liquid will invade the detection part 116, but the optical design is implemented assuming the case of measurement in a state where the light passing part is filled with the liquid.

[0081] According to the structure of the present invention, most of the capillary containing the detection part 116 can be integrally temperature-adjusted. In electrophoresis-based sample separation, the mobility of the sample changes according to the temperature of the separation medium. Therefore, in order to obtain stable measurement results, it is preferable to minimize the temperature distribution and temporal temperature variation of the capillary as much as possible. It is preferable to keep the temperature of the capillary constant not only when the device is in a constant environment but also when the ambient temperature around the device changes.

[0082] However, in conventional capillary electrophoresis devices, there is a problem that spatial distribution and temporal variation of temperature are more likely to occur in the detection part 116 than in other parts of the capillary. In order to perform fluorescence measurement at the detection part 116, it is necessary to irradiate the capillary 111 with excitation light and guide the generated fluorescence to the detector. In order to achieve fluorescence measurement by propagating light in free space, an opening through which the excitation light and fluorescence pass must be provided, and a temperature adjustment mechanism cannot be provided at this opening part. In addition, since the capillary array must be fixed to the optical measurement mechanism inside the device, heat conduction occurs through the fixing part and the temperature changes.

[0083] Regarding the spatial distribution and temporal variation of temperature caused by the influence of the opening, for example, countermeasures such as providing a transparent window with high heat insulation at the opening part and providing a separate temperature adjustment mechanism at the opening part can be implemented. However, in the countermeasure of providing a transparent window, the optical performance deteriorates due to reflection of the window, etc., and there is still non-uniformity of heat conduction due to differences in the material and structure of the window and the surrounding materials and structures. In addition, in the countermeasure of providing a separate temperature adjustment mechanism at the opening part, heating and cooling are performed considering the heat conduction state near the opening, but the structure and control for keeping the temperature uniform with other parts may become complicated. In addition, the situation where the opening part has more heat exchange with the outside than other parts remains unchanged, so there remains a problem that the capillary temperature is likely to change due to temperature changes in the external environment.

[0084] On the other hand, in the structure of the present disclosure, excitation light and fluorescence are exchanged via optical fibers, so the measurement window part can be substantially completely isolated from the outside. On this basis, if through Figure 7If the temperature of the entire capillary 111 including the detection part 116 is adjusted by the method or the like shown in the figure, the sample injection end of the capillary that has to be connected to the outside structurally and all parts other than the connection part to the pump unit 105 can be uniformly temperature-adjusted as a whole. In addition, there is no need to provide a special mechanism for temperature adjustment, and stable temperature control can be easily performed.

[0085] Moreover, the positions of the capillary 111, the cartridge-side irradiation optical fiber 114, and the cartridge-side detection optical fiber 115 are fixed inside the detection part 116. Therefore, it is not necessary to firmly fix the entire capillary 111 including the detection part 116 to the capillary cartridge 110 or the electrophoresis apparatus 100. Therefore, the detection part 116 can be fixed in such a way that it hardly contacts the housing of the capillary cartridge 110 in order to reduce heat transfer to the peripheral part, or a heater and a heat insulating material can be provided around the entire capillary 111 including the detection part 116.

[0086] As a result, it is possible to control the entire capillary 111 to a uniform temperature without providing a separate heat insulating mechanism and temperature control mechanism in the measurement window part. In addition, the heat transfer path to the outside can be reduced, and electrophoresis-based sample separation can be more stably performed even if there is a temperature change outside the apparatus.

[0087] <Example 1: Summary>

[0088] The electrophoresis apparatus 100 of the present Embodiment 1 includes an irradiation optical system 101 and a main body side irradiation optical fiber 112, a detection optical system 102 and a main body side detection optical fiber 113. The capillary cartridge 110 includes a cartridge side irradiation optical fiber 114 and a cartridge side detection optical fiber 115. The relative positions of the capillary 111, the cartridge side irradiation optical fiber 114, and the cartridge side detection optical fiber 115 are adjusted and fixed at the detection part 116. The main body side irradiation optical fiber 112 is connected to the cartridge side irradiation optical fiber 114, and the main body side detection optical fiber 113 is connected to the cartridge side detection optical fiber 115 through an optical fiber connector 117. With the above structure, it is possible to achieve both robustness against vibration and external environment changes and easy replaceability of the capillary. In addition, by adjusting the temperature of the entire capillary 111 including the detection part 116 as a whole, the temperature distribution of the capillary can be made uniform, and temperature fluctuations can be reduced.

[0089] <Embodiment 2>

[0090] Figure 8This is a structural diagram of the electrophoresis apparatus 800 according to Embodiment 2 of the present disclosure. The components of the electrophoresis apparatus 800 according to Embodiment 2 are the same as those of the electrophoresis apparatus 100 according to Embodiment 1. However, it is different from Embodiment 1 in that consumables such as polymers, buffers, and cleaning liquids and the flow path structure for flowing them are integrated with the capillary cartridge 801. Similarly to Embodiment 1, the capillary cartridge includes a capillary 111, a cartridge-side illumination optical fiber 114, a cartridge-side detection optical fiber 115, and a detection part 116 inside. A sample injection-side flow path 802 is provided at the sample injection end of the capillary 111. A solution tank 803 and a waste liquid tank 804 are connected to the sample-side injection flow path 802. An electrode 127 is provided in the sample injection-side flow path 802. Buffers, cleaning liquids, etc. are stored in the solution tank. The end of the capillary 111 on the opposite side is connected to a polymer injection flow path 805. A polymer container 104 and a buffer container 107 are connected to the polymer injection flow path 805. According to this structure, the maintenance operations performed by the user are concentrated on the replacement of the capillary cartridge 801, and compared with the structure of Embodiment 1, the maintenance man-hours can be reduced.

[0091] In Embodiment 2, the same operations as those in Embodiment 1 ( Figure 2 ) are performed by flowing each liquid through the flow path. The polymer injection flow path 805 has a structure similar to the pump unit 105 and performs polymer injection into the capillary 111 (S202 to S206). In addition, in the present embodiment, a drive unit 806 that drives a mechanism equivalent to the syringe 119 is provided on the electrophoresis apparatus 800 side.

[0092] The cleaning of the capillary (S207, S210, S213) and the electrical connection between the front end of the capillary and the electrode by injecting the buffer (S208, S214) are implemented by delivering the cleaning liquid and the buffer from the solution tank to the sample injection-side flow path 802. Each solution is stored in a syringe, and the liquid can be delivered by pushing the syringe, or a mechanism for delivering the liquid can be assembled separately. Similar to the structure for polymer injection, a liquid delivery unit 807 that supplies power for liquid delivery is provided on the electrophoresis apparatus 800 side. In addition, the waste liquid is discarded into the waste liquid tank 804.

[0093] Regarding the injection of the sample (S212), it is implemented by injecting the sample from the outside into the sample injection side flow path 802. The sample is held in the sample cassette 808, and the sample cassette control unit 809 injects the sample into the sample injection side flow path 802. In addition, the sample cassette 808 can only temporarily hold the sample introduced by the user and convey the sample to the sample injection side flow path 802 at the timing of sample injection (S212). It can also perform pretreatment such as purification of the sample and mixing with reagents in addition to liquid feeding. When pretreatment is also performed in the sample cassette 808, the sample cassette control unit 809 controls all processes such as liquid feeding and mixing of various required reagents. For example, Patent Document 3 discloses an apparatus and its structure for consistently performing pretreatment of a sample to analysis based on capillary electrophoresis.

[0094] In the electrophoresis apparatus 800 of Embodiment 2, since consumables are built in the capillary cassette 801, it is impossible to perform operations such as individually replacing each consumable such as a capillary, polymer, and buffer according to the consumption state. On the other hand, the ease of device maintenance including consumable replacement is emphasized. Such a structure is particularly suitable for use by users who are not familiar with device operation. Therefore, for the replacement of the capillary cassette 801, it is also expected that it can be simply implemented without special operations.

[0095] Figure 9 An example of the structure in which the capillary cassette 801 is provided in the electrophoresis apparatus 800 of Embodiment 2 is shown. Figure 9 (a) shows the electrophoresis apparatus 800, the capillary cassette 801, the sample cassette 808, and the control device 109. In this structure, the capillary cassette 801 is connected in a form inserted into the capillary cassette insertion portion 901 provided in the electrophoresis apparatus 800. The sample cassette 808 is connected in a form inserted into the sample cassette insertion portion 902 provided in the electrophoresis apparatus 800. Figure 9 The control device 109 in (a) only shows the screen, but it can also be a tablet PC, a notebook PC, a desktop PC, etc., and can also be integrally assembled inside the electrophoresis apparatus 800.

[0096] Figure 9 (b) shows an example of the structure of the capillary cassette 801. The functions and operations of each component are as described above. In this example, the temperature adjustment of the capillary is implemented by the heater 903. In Figure 9 In (b), the heater 903 is provided under the capillary 111, but in order to improve the temperature adjustment accuracy, it can also be configured in a form sandwiching the capillary 111, or a heat insulating material can be provided on the upper surface of the capillary 111.

[0097] In Figure 9In the example of (b), the electrical connection and the optical connection are performed by the connector 904. An electrical connector and an optical connector are built in the connector 904. In addition, in the case where the temperature of the capillary is adjusted by the circulation of a fluid, a connector for connecting the flow path is provided. Regarding the supply of external force from the drive unit 806 and the liquid feeding unit 807, for example, it is implemented by applying a force to the liquid feeding mechanism 905 from above the cassette by a mechanical mechanism provided in the electrophoresis apparatus 800. As an example, the liquid feeding mechanism 905 has a syringe-like structure and moves up and down through the electrophoresis apparatus 800 to feed the liquid. In addition, the valve 906 is also opened and closed by the mechanical force from the electrophoresis apparatus 800. In addition, these mechanical forces are not limited to vertical movement, and a rotational force or the like may be applied. In addition, the case where components that generate mechanical forces such as a motor and a solenoid are provided inside the capillary cassette 801 is not excluded.

[0098] Through Figure 9 With the structure shown, the user can easily attach and detach the sample cassette 801 to and from the electrophoresis apparatus 800. When the sample cassette 801 is inserted deep into the capillary cassette insertion portion 901 of the electrophoresis apparatus 800, the sample cassette 801 and the electrophoresis apparatus 800 are electrically and optically connected through the connector 904. The mechanism for inserting mechanical force into the liquid feeding mechanism 905, the valve 906, etc. approaches from above the sample cassette 801. The mechanical mechanism is arranged so as not to cause interference when attaching and detaching the sample cassette 801, or to move to a position where no interference is caused during attachment and detachment. The sample cassette 801 is a consumable and needs to be newly replaced with a new one after being used a certain number of times, so it has an advantage in terms of easy attachment and detachment. In addition, through the optical connection using an optical fiber and an optical connector, as described above, the optical unit becomes robust against vibrations and changes in the external environment.

[0099] When the sample cassette 801 is attached to the electrophoresis apparatus 800, it is mechanically fixed to the electrophoresis apparatus 800 so as not to fall off. This fixing mechanism is provided in the housing portion of the sample cassette 801. On the other hand, a fixing mechanism for fixing to an adapter is usually built in a commercially available optical connector. The fixing mechanism of the sample cassette 801 and the fixing mechanism of the optical connector can be fixed simultaneously when attached to the electrophoresis apparatus 800. Alternatively, the fixing mechanism may not be built in the optical connector, and only the fixing mechanism of the sample cassette 801 may be used for fixing.

[0100] As described above, the optical system of the electrophoresis apparatus 800 of the present disclosure is robust against vibrations and external environmental changes by using optical fibers and optical connectors. However, when a strong impact is applied to the apparatus, the positions of the optical components change and the components may be damaged. As protection against strong impacts, there is a method of mounting a buffer material on the object to be protected. In a structure in which light beams propagate in free space, the relative positions of the excitation light irradiation optical system, the capillary, and the detection optical system need to be fixed. Therefore, for example, it is considered to mount the excitation light irradiation optical system, the capillary, and the detection optical system on the same structural support, and protect the above structural support with a buffer material. In this case, the object to be protected includes a light source, a detection system, a structural support, etc., and has a corresponding weight. It is necessary to provide a damping mechanism that supports this weight and has sufficient damping performance.

[0101] On the other hand, in the structure of the present disclosure, the irradiation optical system 101, the detection optical system 102, and the detection part 116 are connected by optical fibers. Since the optical fiber has flexibility, it is possible to separately protect the irradiation optical system 101, the detection optical system 102, and the detection part 116 with a buffer material. In particular, the detection part 116 includes a capillary 111 from which the coating has been removed, a cassette-side irradiation optical fiber 114, and a cassette-side detection optical fiber 115. The positional accuracy among the three is important for the measurement performance, and therefore protection against impact is particularly required.

[0102] As Figure 10 shown, the detection part 116 can be fixed to a structural support 1001 such as the inner wall of the capillary cassette 801 via a buffer material 1002. The detection part 116 only includes lightweight components such as the capillary 111, the cassette-side irradiation optical fiber 114, and the cassette-side detection optical fiber 115, and is lightweight. Therefore, by using only a soft component such as rubber as the buffer material 1002, sufficient damping performance can be obtained with a simple structure such as bonding and fixing the structural support 1001, the buffer material 1002, and the detection part 116.

[0103] In the structure disclosed in Patent Document 2, a detection optical fiber array is connected to a capillary array included in a capillary cassette. In this way, even in a form in which a connection mechanism including an optical fiber is connected to the capillary inside the cassette, it is possible to protect the cassette and the connection mechanism from the influence of impact in a state where they are integrated. However, the structure of the present disclosure can also reduce the weight of the detection part 116 compared with the above structure. This is advantageous for protection against vibration and impact.

[0104] <Example 2: Summary>

[0105] The electrophoresis device 800 of the second embodiment has components similar to those of the electrophoresis device 100 of the first embodiment and performs similar operations, but is different in that consumables such as polymers and buffers are provided inside the capillary cassette 801. In the capillary cassette 801, in addition to the optical connector, an electrical connector and a fluid connector are also provided. When the capillary cassette 801 is inserted into the electrophoresis device 800, the two are connected through these connectors. In addition, mechanical force is supplied from the electrophoresis device 800. The detection part 116 is fixed to the structural support 1001 via the buffer material 1002.

[0106] <Embodiment 3>

[0107] Examples of detecting a sample in a capillary by fluorescence measurement are shown in the first and second embodiments, but the structure of the present disclosure also functions in methods other than fluorescence detection. As an example, the case of detecting a sample by measuring light absorbance is described in the third embodiment. The structure and operation of the electrophoresis device in the third embodiment are the same as those in the first and second embodiments, and thus are omitted. Since the light detection method in the third embodiment is different from those in the first and second embodiments, the structure of the detection part 116 is different.

[0108] Figure 11 The details of the detection part 116 in the third embodiment are shown. In the measurement of light absorbance, it is necessary to irradiate the capillary 111 with light emitted from the cassette-side irradiation optical fiber 114 and collect it by the cassette-side detection optical fiber 115 after passing through the capillary 111. In the present embodiment, this is achieved by arranging the cassette-side irradiation optical fiber 114 and the cassette-side detection optical fiber 115 so as to face each other with the capillary 111 therebetween.

[0109] Figure 11 (a) of shows the structure of the fixing substrate 1101 for fixing the capillary 111, the cassette-side irradiation optical fiber 114, and the cassette-side detection optical fiber 115 on the substrate for adopting the above structure. The capillary fixing groove 1102, the irradiation optical fiber fixing groove 1103, and the detection optical fiber fixing groove 1104 are provided on the fixing substrate 1101. Figure 11 (b) of is a view in which the capillary 111, the cassette-side irradiation optical fiber 114, and the cassette-side detection optical fiber 115 are fixed on the fixing substrate 1101.

[0110] In addition, in the present embodiment, it is configured not to use optical elements other than optical fibers, but optical elements other than optical fibers may be provided on the fixing substrate 1101 as needed. In addition, in the case of providing a plurality of capillaries, the Figure 11 shown structure may be arranged in parallel, or a structure similar to Figure 3 、 Figure 4 may be adopted to arrange the irradiation and detection optical fiber arrays so as to face each other with the capillary array therebetween.

[0111] In addition, it is also possible to take into account the structure for measuring light absorbance and the structure for fluorescence measurement. For example, as Figure 12 shown, for the capillary 111, an irradiation optical fiber 1201 for absorbance measurement and a detection optical fiber 1202 for absorbance measurement are provided, and the fluorescence irradiation optical fiber 1203 and the fluorescence detection optical fiber 1204 are arranged at an angle of 45 degrees with respect to the irradiation optical fiber 1201 for absorbance measurement and the detection optical fiber 1202 for absorbance measurement.

[0112] <Example 3: Summary>

[0113] In the detection part 116 of the present Embodiment 3, the cartridge-side irradiation optical fiber 114 and the cartridge-side detection optical fiber 115 are fixed opposite to each other with the capillary 111 interposed therebetween. The light emitted from the cartridge irradiation optical fiber 114 passes through the capillary 111 and then enters the cartridge-side detection optical fiber 115, thereby performing absorbance measurement.

[0114] <Embodiment 4>

[0115] In Embodiment 4, regarding the fixing substrate for fixing the capillary, the irradiation optical fiber, and the detection optical fiber, a method of forming a through hole by forming grooves on both the front and back surfaces of the substrate and the structure of the substrate manufactured by this method will be described.

[0116] Figure 13 The structure of the fixing substrate 1301 showing Embodiment 4 is shown. Figure 13 (a) of shows the front surface of the fixing substrate 1301, Figure 13 and (b) of shows the back surface of the fixing substrate 1301. The fixing substrate 1301 is the same as the fixing substrate 301 of Embodiment 1, and fixes four capillaries, one irradiation optical fiber, and four detection optical fibers. On the front surface of the fixing substrate 1301, there are provided a capillary fixing groove 1302 for positioning four capillaries and an irradiation optical fiber fixing groove 1303 for positioning one irradiation optical fiber. On the back surface, there is provided a through hole forming groove 1304 formed in a direction orthogonal to the capillary fixing groove 1302. A through hole 1305 is formed at the intersection of the capillary fixing groove 1302 and the through hole forming groove 1304. Four detection optical fibers are positioned by being inserted into the through hole 1305.

[0117] The positioning method of the capillary, the irradiation optical fiber, and the detection optical fiber is similar to the positioning method in the substrate described in (e) of Embodiment 1. However, in the substrate described in (e) of Figure 4 , after the capillary fixing groove is formed, the through hole is formed by other means. In contrast, the difference between the substrate of the present embodiment and that is that the fixing groove and the through hole are formed simultaneously by forming grooves on both the front and back surfaces. Figure 4

[0118] ​The method for forming the through-hole will be described in detail below. The capillary fixing groove 1302 and the through-hole forming groove 1304 are each formed to a depth that does not penetrate the substrate (groove depth < substrate thickness). On the other hand, it is set that the sum of the depth of the capillary fixing groove 1302 and the depth of the through-hole forming groove 1304 is greater than the thickness of the substrate. At this time, the bottom of the capillary fixing groove 1302 overlaps with the through-hole forming groove 1304, and the overlapping part of the grooves is connected to form a through-hole 1305 that penetrates the substrate. By adopting this method, it is possible to simultaneously form the grooves and form the through-holes at positions along the grooves.

[0119] When manufacturing the fixed substrate 1301, as an example, the substrate material may be silicon, and grooves having a V-shaped cross-section may be formed by anisotropic etching of silicon. First, thermal oxide films are formed on the front and back surfaces of a silicon substrate having a 100 plane. Then, after applying a resist, performing exposure and development, etching is performed with hydrofluoric acid to remove the oxide film in the portion where the V-grooves are to be formed. That is, it becomes a state where the oxide film in the portions corresponding to the capillary fixing groove 1302, the irradiated optical fiber fixing groove 1303, and the through-hole forming groove 1304 has been removed. Then, anisotropic etching of the substrate is performed using an alkaline solution such as an aqueous potassium hydroxide solution to form V-grooves. The capillary fixing groove 1302, the irradiated optical fiber fixing groove 1303, and the through-hole forming groove 1304 are formed by etching. The through-hole 1305 is formed at the time point when the bottom of the capillary fixing groove 1302 intersects the bottom of the through-hole forming groove 1304. The size of the through-hole can be adjusted by adjusting the width of the through-hole forming groove 1304 and the etching time. After forming the V-grooves, the remaining oxide film is removed. In addition, the material of the mask used in the anisotropic etching, the solution used in the etching, etc. may also be different from the above.

[0120] In the structure in which laser is irradiated from the side of a capillary array in which capillaries are arranged in a single row to excite the phosphor in the capillaries, the distances of the irradiated optical fibers and the capillaries from the substrate surface need to be made consistent with an accuracy of about 10 μm or less. If it is anisotropic etching of silicon, the width and angle of the V-grooves can be controlled with high precision, and the heights of the capillaries and the irradiated optical fibers from the substrate surface can be made consistent with high precision.

[0121] In addition, the material of the substrate of Embodiment 4 does not have to be silicon, and the method for forming the fixing grooves does not have to be anisotropic etching. As long as the grooves for fixing the capillaries, the irradiated optical fibers, and the detection optical fibers can be formed with sufficient accuracy, and as long as the grooves can be formed to a depth such that through-holes can be formed in the substrate by etching from the front and back surfaces.

[0122] Figure 14Figure (a) shows a structural diagram when the capillary 1401, the irradiation optical fiber 1402, and the detection optical fiber 1403 are installed on the fixed substrate 1301. Figure 14 Figure (b) shows a cross-sectional view obtained by cutting the structure of Figure (a) along a plane perpendicular to the capillary fixing groove 1302 at the position of the through hole. Figure 14 Figure (a)

[0123] The capillary 1401 is fixed through the capillary fixing groove 1302 such that the central axis of the capillary 1401 is located at a position away from the surface of the fixed substrate 1301 by a certain distance. This can be achieved, for example, by fixing the capillary 1401 with an adhesive or the like while pressing the capillary 1401 against the capillary fixing groove 1302 using a capillary pressing substrate (not shown).

[0124] In Figure 14 Figure (a), the irradiation optical fiber 1402 is fixed to the fixed substrate 1301 in a state of being inserted into the position adjustment member 1404. The position adjustment member 1404 is a cylindrical member having a hole at its center for inserting the irradiation optical fiber 1402. At the end of the position adjustment member 1404 on the capillary side, there is a portion for inserting a lens, and this lens is used to collimate the excitation light emitted from the irradiation optical fiber 1402. The optical axis adjustment between the irradiation optical fiber 1402 and the lens is performed by inserting the lens into this portion. As an example, a conical hole is provided at the end of the position adjustment member 1404, and a spherical lens is inserted into the conical hole and the end of the spherical lens is fixed with an adhesive or the like. Regarding the distance between the irradiation optical fiber 1402 and the lens, it can be adjusted while irradiating light from the irradiation optical fiber 1402 and observing the shape of the light spot imaged by the lens. In addition, when the number of capillaries 1401 is one, it is not necessary to use a lens. Also, the fixing of the lens does not necessarily need to be performed by the position adjustment member 1404, and methods such as providing a recess for installing the lens on the fixed substrate 1301 can also be adopted.

[0125] The detection optical fiber 1403 is fixed by being inserted into the through hole 1305. At this time, as shown in Figure 14 Figure (b), the excitation light 1405 and the detection optical fiber 1403 are arranged orthogonally.

[0126] In the case where the number of capillaries in the fixed substrate of Embodiment 4 is multiple, it also has the effect of reducing crosstalk between capillaries. When the number of capillaries is multiple, there is a case where fluorescence emitted from a certain capillary enters an optical fiber for detecting a different capillary. In such a case, the fluorescence of a certain capillary is misrecognized as fluorescence emitted from another capillary (crosstalk). In the presence of crosstalk, the fluorescence caused by the component a of the sample A analyzed by a certain capillary is misrecognized as a signal from a capillary analyzing another sample B, which may be the main cause of an incorrect analysis result such that the component a is included in the sample B.

[0127] Crosstalk between capillaries can occur, for example, in the path shown in Figure 15 (a). In Figure 15 (a), the light rays (arrows) of fluorescence generated by the capillary on the left are reflected on the surface of the capillary on the right and enter the detection optical fiber for detecting the capillary on the right. The generation of crosstalk in the above path is suppressed by the structure of the fixed substrate of Embodiment 4. In the fixed substrate of Embodiment 4, the region other than the part where the groove for fixing the capillary is formed becomes a wall that separates the capillaries. As shown in Figure 15 (b), the generation path of crosstalk shown in Figure 15 (a) is blocked by this wall.

[0128] Figure 16 shows the simulation results of the crosstalk suppression effect of the fixed substrate. In this simulation, 4 capillaries with an inner diameter of 50 μm and an outer diameter of 343 μm were arranged at intervals of 1 mm, and fluorescence was detected by an optical fiber with a core diameter of 200 μm and NA 0.5. In this simulation, a region with a length of 50 μm in the inner diameter part of 1 out of 4 capillaries was made to emit light, and the ratio of the fluorescence incident on the detection optical fiber of other capillaries, that is, the ratio of crosstalk, was calculated.

[0129] Figure 16 (a) shows the value of crosstalk without the fixed substrate. The horizontal axis of the graph represents the optical fiber that emits light, and each bar graph shows the crosstalk observed in the optical fiber for detecting the capillary that emits light. In the absence of the fixed substrate, crosstalk of about 0.08% was observed in the optical fiber for detecting the capillary adjacent to the capillary that emits light. On the other hand, Figure 16 (b) shows the crosstalk with the fixed substrate. In the presence of the fixed substrate, the observed crosstalk is about 0.002%, and it can be seen that the ratio of crosstalk is reduced to about 1 / 40.

[0130] <Example 4: Summary>

[0131] In the fixed substrate 1301 of Embodiment 4, a capillary fixing groove 1302 and an irradiation optical fiber fixing groove 1303 are provided on the surface of the substrate, and a through-hole forming groove 1304 is provided on the back surface. A through-hole 1305 is formed at the intersection of the capillary fixing groove 1302 and the through-hole forming groove 1304. The region where the capillary fixing groove 1302 is not formed becomes a wall that separates adjacent capillaries, reducing crosstalk between the capillaries.

[0132] <Regarding Modification Examples>

[0133] The present disclosure is not limited to the above-described embodiments, and includes various modification examples. For example, the above-described embodiments are embodiments described in detail for easy understanding of the present disclosure, and are not necessarily limited to having all the structures described. In addition, a part of the structure of a certain embodiment can be replaced with the structure of another embodiment, and in addition, the structure of another embodiment can be added to the structure of a certain embodiment. In addition, with respect to a part of the structure of each embodiment, addition, deletion, and replacement of other structures can be performed.

[0134] Description of Reference Numerals

[0135] 100 Electrophoresis device

[0136] 101 Irradiation optical system

[0137] 102 Detection optical system

[0138] 103 High-voltage power supply

[0139] 104 Polymer container

[0140] 105 Pump unit

[0141] 106 Temperature control device

[0142] 107 Buffer solution container

[0143] 108 Automatic sampling unit

[0144] 109 Control device

[0145] 110 Capillary box

[0146] 111 Capillary

[0147] 112 Main body side irradiation optical fiber

[0148] 113 Main body side detection optical fiber

[0149] 114 Cartridge side irradiation optical fiber

[0150] 115 Cartridge side detection optical fiber

[0151] 116 Detection site

[0152] 117 Fiber Optic Connector

[0153] 118 Flow Path Block

[0154] 119 Syringe

[0155] 120 Check Valve

[0156] 121 Valve

[0157] 122 Buffer Solution Tray

[0158] 123 Cleaning Water Tray

[0159] 124 Waste Liquid Tray

[0160] 125 Sample Tray

[0161] 126 Stage

[0162] 127 Electrode

[0163] 301 Fixed Substrate

[0164] 302 Capillary Tube Fixing Groove

[0165] 303 Fiber Optic Fixing Groove

[0166] 304 Lens Fixing Groove

[0167] 305 Through Hole

[0168] 306 Ball Lens

[0169] 401 Pressing Substrate

[0170] 402 Detection Fiber Optic Array Mounting Hole

[0171] 403 Detection Fiber Optic Array

[0172] 404 Array Fixing Component

[0173] 501 Fixed Substrate

[0174] 502 Capillary Tube Fixing Groove

[0175] 503 Irradiation Fiber Optic Fixing Groove

[0176] 504 Ball Lens Fixing Hole

[0177] 505 Detection Fiber Optic Fixing Groove

[0178] 601 Assembly Component

[0179] 602 Electrical Connector

[0180] 603 Temperature Regulation Connector

[0181] 701 Chip Heater

[0182] 702 Fluid supply port

[0183] 703 Fluid discharge port

[0184] 704 Partition wall

[0185] 800 Electrophoresis device

[0186] 801 Capillary cassette

[0187] 802 Sample injection side flow path

[0188] 803 Solution tank

[0189] 804 Waste liquid tank

[0190] 805 Polymer injection flow path

[0191] 806 Driving unit

[0192] 807 Liquid feeding unit

[0193] 808 Sample cassette

[0194] 809 Sample cassette control unit

[0195] 901 Capillary cassette insertion part

[0196] 902 Sample cassette insertion part

[0197] 903 Heater

[0198] 904 Connector

[0199] 905 Liquid feeding mechanism

[0200] 906 Valve

[0201] 1001 Structural support

[0202] 1002 Buffer material

[0203] 1101 Fixed substrate

[0204] 1102 Capillary fixing groove

[0205] 1103 Irradiation optical fiber fixing groove

[0206] 1104 Detection optical fiber fixing groove

[0207] 1201 Irradiation optical fiber for absorbance measurement

[0208] 1202 Detection optical fiber for absorbance measurement

[0209] 1203 Irradiation optical fiber for fluorescence measurement

[0210] 1204 Detection optical fiber for fluorescence measurement

[0211] 1301 Fixed substrate

[0212] 1302 Capillary fixing groove

[0213] 1303 Irradiation optical fiber fixing groove

[0214] 1304 Through-hole forming groove

[0215] 1305 Through-hole

[0216] 1401 Capillary

[0217] 1402 Irradiation optical fiber

[0218] 1403 Detection optical fiber

[0219] 1404 Position adjustment component

[0220] 1405 Excitation light.

Claims

1. A capillary electrophoresis device, which is an electrophoresis device having a light source, a first irradiation optical fiber that guides light from the light source, a detector that detects light, and a first detection optical fiber that guides light to the detector, Characterized in that, By connecting the first irradiation optical fiber and the second irradiation optical fiber and connecting the first detection optical fiber and the second detection optical fiber, a capillary cartridge having a capillary, the second irradiation optical fiber, and the second detection optical fiber is installed on the electrophoresis device, In the capillary cartridge, the optical axes of the second irradiation optical fiber and the second detection optical fiber are fixed so as to intersect in the inner cavity of the capillary.

2. The capillary electrophoresis device according to claim 1, characterized in that, The detection method of the sample is fluorescence measurement.

3. The capillary electrophoresis device according to claim 1, characterized in that, The detection method of the sample is light absorbance measurement.

4. The capillary electrophoresis device according to claim 2, characterized in that, The capillary cartridge has a plurality of the capillaries, For the capillaries arranged in a row on the substrate, excitation light is incident on the side surface of the array formed by the capillaries arranged in a row through the second irradiation optical fiber, The fluorescence from each capillary captured by a plurality of the second detection optical fibers is guided to the detector through the first detection optical fiber.

5. The capillary electrophoresis device according to claim 2 or 4, characterized in that, In the capillary cartridge, the substrate on which the capillary and the second irradiation optical fiber are fixed and the substrate on which the second detection optical fiber is fixed are fixed to be substantially perpendicular.

6. The capillary electrophoresis device according to claim 2 or 4, characterized in that, The second detection optical fiber is fixed on the same substrate so as to be substantially perpendicular to the plane formed by the capillary and the second irradiation optical fiber, The alignment of the three is performed through grooves and through holes formed on the substrate.

7. The capillary electrophoresis device according to claim 2 or 4, characterized in that, In the capillary cartridge, the capillary, the second irradiation optical fiber, and the second detection optical fiber are fixed on the substrate so as to all be in the same plane, The position adjustment among the three is performed through grooves formed on the substrate.

8. The capillary electrophoresis device according to claim 1, characterized in that, The capillary and the second irradiation optical fiber, the second detection optical fiber, and the fixing member that fixes the second irradiation optical fiber, the second detection optical fiber, and the capillary are integrally temperature-controlled.

9. The capillary electrophoresis device according to claim 1, characterized in that, The capillary is provided in a housing and is connected to the electrophoresis device via an optical connector provided on the outer wall of the housing.

10. The capillary electrophoresis device according to claim 9, characterized in that, The portion where the optical axes of the second irradiation optical fiber and the second detection optical fiber are fixed so as to intersect in the inner cavity of the capillary is fixed to the housing via a buffer structure.

11. A capillary cartridge, which has a capillary, a first irradiation optical fiber, and a first detection optical fiber, characterized in that, Fix the optical axes of the first irradiation optical fiber and the first detection optical fiber so that they intersect inside the capillary lumen. Install the capillary cassette on the electrophoresis device by connecting the first irradiation optical fiber to the second irradiation optical fiber and the first detection optical fiber to the second detection optical fiber. Among them, the electrophoresis device includes: a light source, the second irradiation optical fiber that guides light from the light source, a detector that detects light, and the second detection optical fiber that guides light to the detector.

12. A capillary cassette, characterized in that When setting the capillary cassette on the electrophoresis device, connect the first irradiation optical fiber to the second irradiation optical fiber and the first detection optical fiber to the second detection optical fiber by inserting the capillary cassette into the insertion part of the electrophoresis device.

13. The capillary electrophoresis device according to claim 6, characterized in that The substrate has a groove for fixing the capillary and a groove for fixing the second irradiation optical fiber on its surface, a through-hole forming groove orthogonal to the groove for fixing the capillary on its back surface, and the second detection optical fiber is provided in the through-hole formed at the intersection of the groove for fixing the capillary and the through-hole forming groove.

14. The capillary electrophoresis device according to claim 6, characterized in that The substrate has a structure for blocking light between the through-holes.

15. The capillary cassette according to claim 12, characterized in that Fix the capillary, the second irradiation optical fiber, and the second detection optical fiber to the substrate. The substrate has a groove for fixing the capillary and a groove for fixing the second irradiation optical fiber on its surface, a through-hole forming groove orthogonal to the groove for fixing the capillary on its back surface, and the second detection optical fiber is provided in the through-hole formed at the intersection of the groove for fixing the capillary and the through-hole forming groove.

16. A method for manufacturing a fixed substrate for fixing a substrate, the fixed substrate fixing a capillary, a second irradiation optical fiber connected to a first irradiation optical fiber, and a second detection optical fiber connected to a first detection optical fiber in a capillary cassette connected to an electrophoresis device. Among them, The electrophoresis device includes: a light source, the first irradiation optical fiber that guides light from the light source, a detector that detects light, and the first detection optical fiber that guides light to the detector. It is characterized in that Form a groove for fixing the capillary and a groove for fixing the second irradiation optical fiber on the surface of the fixed substrate, form a groove orthogonal to the groove for fixing the capillary on the back surface, and intersect the groove for fixing the capillary with the orthogonal groove to form a through-hole for fixing the second detection optical fiber.

17. The method for manufacturing a fixed substrate according to claim 16, characterized in that The material of the fixed substrate is silicon, and the groove for fixing the capillary, the groove for forming the irradiation optical fiber, and the groove orthogonal to the groove for fixing the capillary are formed by anisotropic etching.

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