JP3325257B2 - Capillary electrophoresis device - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an apparatus suitable for electrophoretic analysis of biological substances such as DNA, RNA, and protein, and more particularly, to an apparatus using a fluid separation medium such as a fluid gel or a fluid polymer as a separation medium. The present invention relates to an electrophoretic separation device having a fluorescence detection function.

[0002]

2. Description of the Related Art DNA analysis technology has made remarkable progress with the practical use of a real-time fluorescence detector. Conventionally, plate-type gel electrophoresis devices have been used for DNA analysis, but demand for high-speed and high-throughput devices such as genomic analysis has increased.
A so-called capillary gel electrophoresis apparatus, in which a capillary is filled with a gel or a flowable polymer and used as an analysis tube, has become widespread. Then, a fluid gel or a fluid polymer has been used as a separation medium, and a fully automatic capillary gel electrophoresis apparatus for automatically refilling the gel as necessary has been developed and put into practical use.

[0003]

Although these devices have been accepted in the world as second-generation DNA fluorescence analyzers or DNA sequencers, the analyzer tubes using capillaries are large in spite of their small size. There are also problems such as high prices. On the other hand, with the progress of the genome project, DNA analysis has been performed on a large scale,
The direction to make the database more complete and D
There is a growing trend to easily perform NA analysis and use the results in medicine, the environment, engineering, and so on. Currently, the development of the former system is progressing, but the development of simple and inexpensive devices has hardly progressed. An object of the present invention is to provide a capillary electrophoresis apparatus suitable for simple and inexpensive DNA analysis that meets future needs in view of the current state of such electrophoresis apparatuses.

[0004]

In order to achieve the above object, a small and highly sensitive excitation light irradiation and fluorescence detection system is required. In addition, it is necessary to adopt a device configuration in which the capillaries and the like can be easily attached and detached and do not require adjustment, and have a structure in which extra parts are eliminated as much as possible. Therefore, in the present invention,
The electrophoresis / analysis unit consisting of a capillary tube was fixed to a holder with a buffer tank and fitted on the substrate. At the same time, the small laser and the fluorescence detector were fixed on the same substrate to eliminate the need for optical path adjustment as much as possible. A lens for condensing laser light was fixed to the holder with a buffer solution tank, and the laser light was applied to the center of the capillary without any adjustment.

[0005] Further, the distance between the capillary and an injection device for injecting a fluid separation medium such as a fluid gel or a fluid polymer into the capillary is shortened, the pressure loss is minimized, and a simple pump or manual pump is used. The structure was such that the fluid separation medium in the capillary could be replaced by a vessel.
At the time of gel injection, the valve at the capillary end in contact with the buffer tank is usually switched to the gel injection side.However, for the sake of compactness and simplicity, the capillary end is connected to the fluid separation medium injection side, and the capillary end is connected. A structure was adopted in which a pore was provided near the end, or a structure was used in which a capillary was connected to a capillary for injecting a fluid separation medium with a porous material to simplify the process.

[0006] Fluorescence detection is performed using a rotating filter. To reduce the size of the filter, the structure is such that fluorescence is first condensed and then passed through the filter when the fluorescence beam has a small cross section. The rotation filter is rotated at a low speed by a simple motor. By monitoring the rotation position of the filter using a light-emitting element and a light detection diode, the detection time and timing of the fluorescence transmitted through a plurality of (for example, four) filters are determined. I adjusted it.

When a plurality of capillaries are used, an irradiation laser beam is scanned in synchronization with a rotary filter, and a fluorescence signal obtained from each capillary is detected as a signal for each phosphor by time division. Alternatively, all capillaries are simultaneously irradiated from the side, a fluorescent image based on the DNA band migrating in each capillary is formed by a lens, and signals from the respective image points are detected by time division, or a CCD camera is used. A simple system is realized by detecting by receiving light.

That is, a capillary electrophoresis apparatus according to the present invention comprises a main body having a fluorescence excitation light source and a fluorescence detection section and having a capillary holder mounting section, a separation capillary held horizontally and a buffer for immersing the end of the capillary. It has a pair of buffer tanks for storing liquids, includes a capillary holder that can be attached to and detached from the capillary holder mounting part of the main body while holding the capillary, and emits fluorescence with the capillary holder attached to the capillary holder mounting part of the main body The capillary is irradiated with excitation light from an excitation light source, and the fluorescence generated from a sample in the capillary is detected by a fluorescence detection unit.

[0009] The capillary holder can be provided with a lens for converging excitation light and / or an optical lens for condensing fluorescence. By adopting such a structure, the electrophoretic analysis can be performed without adjusting the excitation light irradiation optical system and the fluorescence detection optical system only by attaching the capillary holder holding the capillary to the apparatus main body. . The fluorescence detection unit detects a rotational position of a rotating wheel equipped with a plurality of bandpass filters having different transmission wavelength bands, a single photomultiplier tube located behind the rotating wheel, for example, a rotating wheel including a light emitting diode and a photodiode. A single photomultiplier tube can identify and detect a plurality of fluorescences. For example, D
Emission wavelengths from a plurality of fluorescent labels labeled with NA or the like can be identified and detected by selectively receiving light through a band-pass filter of the rotating wheel. Fluorescence identification can also be performed using wavelength dispersion by a diffraction grating or a prism.

[0010] The capillary electrophoresis apparatus according to the present invention also detects light passing through a fluorescence excitation light source, a rotary filter in which a plurality of types of optical filters are separately arranged in a circumferential direction and a radial direction, and an optical filter. A main body having a capillary holder mounting part, a plurality of separation capillaries held in a horizontal direction, and a pair of buffers for containing a buffer solution into which the ends of the capillaries are immersed. And a capillary holder that holds the plurality of capillaries and that can be attached to and detached from the capillary holder mounting part of the main body.The fluorescence excitation light source with the capillary holder mounted on the capillary holder mounting part of the main body Simultaneously irradiate multiple capillaries with the excitation light of In the light detection unit, the fluorescence generated from the sample in the plurality of capillaries is separated and imaged for each capillary at different radial positions of the rotating filter, and the fluorescence generated from each capillary is separated in the circumferential direction. It is characterized in that wavelengths are separated and detected by a plurality of arranged optical filters.

According to one aspect of the capillary electrophoresis apparatus according to the present invention, one end of the capillary has a porous wall portion provided in a buffer tank and is connected to a fluid separation medium injection means. The porous wall of the pipe member allows ions to pass therethrough, but exhibits high resistance to the passage of the fluid separation medium, and the fluid separation medium injection means through the pipe member. It has the function of injecting a fluid separation medium into the capillary. As a fluid separation medium, a fluid gel or a fluid polymer can be used.

According to another aspect of the capillary electrophoresis apparatus according to the present invention, one end of the capillary is fixed to a capillary holder by a jig, and a buffer tank containing a buffer solution for immersing the one end of the capillary is provided with a fluid separation. A movable pipe member is arranged in the direction of the jig provided with a connection portion with the medium injection means, and the fluid separation medium is pressurized in the capillary by bringing the tip of the pipe member into close contact with the jig so as to surround one end of the capillary. Has the ability to inject. After the fluid separation medium is injected under pressure into the capillary, the capillary member can be immersed in the buffer solution in the buffer tank by separating the tube member from the jig. Therefore, electrophoresis can be performed by applying a current in this state.

Further, in the capillary electrophoresis apparatus of the present invention, the sample holding member arranged in the buffer tank with the concave portion holding the sample liquid facing the capillary end, and the sample holding member are moved toward the capillary end. And a function of bringing the sample solution into contact with the end of the capillary and injecting it into the capillary by an electric field. When a cartridge provided with a plurality of sample holding members is mounted, the plurality of samples can be analyzed efficiently by moving the cartridge and sequentially introducing the samples held by each sample holding member from the end of the capillary. Movement of the cartridge, movement and detachment of the sample holding member to the end of the capillary,
By automating the application of a sample injection electric field and the like, a plurality of samples can be automatically analyzed.

According to the present invention, a small, inexpensive and simple DNA
Provided is a capillary electrophoresis device that can be used as an analysis device (DNA checker). That is,
According to the present invention, the DNA was conventionally large and expensive.
Simplified DNA that retains the basic performance of the analyzer and achieves 1/40 in volume, 1/20 in weight, and 1/10 in price, and can be used in any facility without having to worry about the location. An analyzer is provided.

[0015]

Embodiments of the present invention will be described below with reference to the drawings. [Embodiment 1] Gel electrophoresis is widely used for DNA analysis. Capillary gel electrophoresis devices that have been put into practical use recently have features such as easy handling, small amount of consumable sample, and high-speed electrophoresis. (For example, Analytical Chemistry Jan 1
(1999), 31A-37A). In particular, by using a fluid gel instead of a crosslinked gel, exchange of the separation medium is simplified, and full automation of the measurement is achieved, and it is rapidly spreading. However, since the whole apparatus is large-scale, development of a small apparatus utilizing the characteristics of using a capillary is desired. In order to achieve this, a small laser is mounted, a simple and highly sensitive detector is adopted, the optical adjustment of the laser beam convergence position and the capillary center position at the time of laser irradiation is simplified, and a separation medium exchange system is installed. Simplification is key. Here, a small device in which a laser, a detector, and a capillary holder are arranged and fixed on one base is disclosed.

FIG. 1 is a partially cutaway perspective view showing an example of an electrophoresis apparatus according to the present invention. In this apparatus, in order to efficiently arrange optical components, a capillary holder 2 with a buffer tank equipped with a capillary 1 for performing electrophoresis and a laser 3 are arranged on an upper portion of a base 4 of the apparatus main body, and a plurality of types (shown in FIG. A fluorescence detecting unit including a rotating plate 6 with a filter on which color separation filters 5 (in the example, four types) are disposed and a photomultiplier tube 7 is disposed under the base 4 of the apparatus main body.

The laser light emitted from the laser 3 is reflected by the reflection mirror 8, condensed by the irradiation lens 9, and irradiated into the capillary 1. Fluorescence generated from the inside of the capillary 1 by laser light irradiation is efficiently condensed by the condensing lens 10 and is incident on the photomultiplier tube 7 by the coupling lens 13 via the excitation light removing filter 11 and the reflecting mirror 12. Color separation of phosphors of different types (described below with four types of examples) is performed by a filter-equipped rotating plate 6 having four types of color separation filters 5 disposed immediately before a photomultiplier tube 7. The filter-equipped rotary plate 6 is rotated by a rotary motor 14 and enables temporally separating and detecting fluorescence emitted from the inside of the capillary 1. Note that the four types of color separation filters 5 are distinguished from each other by the position detecting cuts 1 provided on the rotating plate 6 with filters.
5 and 16 are detected by the filter position detectors 17 and 18. Of course, other arrangements may be used.

The electrophoresis of the sample is performed in buffer tanks 19 and 2.
This is performed by applying a voltage from the high-voltage power supply 21 to the electrodes 22 and 23 in 0. In the fluorescence detection, a voltage is applied to the photomultiplier tube 7 using a high-voltage power supply 24 for the photomultiplier tube, and the detected fluorescence is converted into a voltage and extracted. In addition, the high voltage power supply 2
The control of 1 and 24, the signal processing of the detection signals of the filter position detectors 17 and 18, and the processing of the signal from the photomultiplier tube 7 are performed by the computer 25.

The present apparatus uses a pen-type laser (for example, a semiconductor laser or a semiconductor-excited YAG laser) 3 as a light source, and uses a photomultiplier tube 7 and a filter-equipped rotating plate 6 in order to reduce costs and achieve miniaturization. Fluorescence detection is performed with the combination of. In addition to these, Capillary 1
Is fixed to the capillary holder 2 with a buffer tank,
The whole is configured by fitting it into the base 4 of the apparatus main body.

Capillary holder 2 with buffer tank
Is composed of an electrophoresis section comprising a capillary 1 for performing electrophoresis and buffer vessels 19 and 20, an optical section comprising an irradiation lens 9 for converging laser light, and a condenser lens 10 for condensing fluorescence. I have. Buffer tank 19,
Electrodes 22 and 23 are provided in 20. The capillary holder 2 with a buffer tank is positioned with a positioning pin or the like, and fitted into a predetermined position of the base 4 with a single touch, thereby irradiating the laser beam emitted from the laser 3 to the center of the capillary 1 accurately. It is designed to efficiently collect fluorescent light from inside. When the capillary holder 2 with a buffer tank is mounted at a predetermined position on the base 4 of the apparatus main body, the electrodes 22 and 23 are connected to a connector on the apparatus main body side by fitting a pin and a socket.

Fluorescence obtained by laser light irradiation is detected from a direction different from the incident direction of the irradiation light (in this example, a direction forming an angle of 90 °). Therefore, the configuration is such that scattered light or the like hardly enters as background light. The area on the substrate occupied by these optical detection components and the electrophoresis section is small, and B5
No. or A4 size can be fixed to all bases. The power supplies need only be equipped with two high voltage modules for electrophoresis and photomultiplier tubes, and are large enough to fit in a storage box of about 8 cm in height with a floor area of the A4 size base.
That is, the capacity of the apparatus is about 5 liters, which is about 1/60 that of the current commercially available product.

FIG. 2 is a conceptual diagram showing a device for injecting a fluid polymer or a fluid gel (fluid separation medium) into the capillary 1 of the capillary holder 2 with a buffer tank, and a sample injecting device. Both sides of the capillary 1 are fixed to the centers of the capillary fixing seal pins 31 and 32. The fluid separation medium is pressed into the capillary 1 from the connector 34 with a fluid separation medium injection nozzle connected to the injector 33, and the connection between the connector 34 and the capillary 1 is performed in the buffer tank 20. The connector 34 with a fluid separation medium injection nozzle is connected to the buffer tank 20.
3A, and is screwed to the wall as shown in FIG.
The fluid separation medium can be introduced into the capillary 1 by screwing the connector 34 with a fluid separation medium injection nozzle and bringing the nozzle tip into close contact with the capillary fixing seal pin 31 fixing the capillary 1. After introducing the fluid separation medium into the capillary 1, FIG.
As shown in (b), when the nozzle of the connector 34 with a fluid separation medium injection nozzle is separated from the seal pin 31 for fixing the capillary, the capillary 1 comes into contact with the buffer solution in the buffer tank 20 and becomes in a state where electricity can be supplied.
Since the length of the capillary 1 is short, the injection of the fluid separation medium can be performed manually without a special pump. The connector 34 with a fluid separation medium injection nozzle has a structure in which the connector main body is fixed to the capillary holder 2 and only the nozzle portion at the tip end opposed to the capillary fixing seal pin 31 is screwed back and forth with respect to the connector main body. Is also good.

The sample is injected by inserting the sample cartridge holder 35 into a predetermined position of the buffer tank 19 using a sample injection tool 37. At that time, the buffer tank 19
Is in a sealed state, and has a structure in which the buffer solution does not leak outside. The sample cartridge holder 35 includes a sample holder 36 and a sample holder cartridge 38 for continuous sample injection.

FIG. 4 shows an outline of the sample injection method. The sample is held in the capillary sample holder 39 in the sample holder 36 in advance, and the sample holder 36 is attached using the sample injector 37 attached to the capillary holder 2 with a buffer tank.
Is moved along the capillary 1 for electrophoresis separation filled with a fluid polymer or a fluid gel, and brought into contact with the end of the capillary 1 to be injected by an electric field. The electric field can be applied, for example, by providing an exposed electrode at the bottom of the capillary-shaped sample holding unit 39 and applying a voltage between the electrode and the electrode 23 in the buffer tank 20 on the opposite side. After the sample is injected, the sample holder 36 is returned to the original position, and the buffer solution is injected into the buffer tank 19, and the sample is subjected to electrophoresis separation.
measure.

It is also possible to fill the buffer tank 19 with a buffer solution before the sample is injected and to inject the sample, if an electric field is applied only between the sample and the capillary at the time of sample injection. This is because, for example, the end of the capillary 1 is inserted halfway into a capillary-shaped sample holder 39 whose diameter is larger than the diameter of the capillary 1, and an electric field due to a voltage applied between the electrodes 22 and 23 is applied to the sample holder 39. This is possible by applying a buffer solution around the inserted capillary 1 so as to be applied between the sample solution and the end of the capillary 1. Furthermore, by enabling sample injection to be performed automatically, fully automatic analysis is possible. For this purpose, as shown in FIG. 2, for example, a sample holder 36 having a plurality of sample holder cartridges 38 for continuous sample injection is prepared, automatically moved for each measurement, and an electric field is applied to sample injection. What is necessary is just to comprise so that it can be performed.

In the case of the apparatus of this example, the length of the capillary 1 is about 20 cm, and the capillary 1 is placed almost horizontally in parallel with the base 4. In this configuration, the effective migration length, that is, the distance between the sample injection part and the measurement window is 15
cm. Since the configuration is such that the total length of the capillary can be shortened, the high-voltage power supply 21 for electrophoresis may be small and 100 V
In order to obtain an electric field strength of / cm, a power supply with an output of 2 KV is sufficient.

Embodiment 2 FIG. 5 is a schematic view showing another example of a capillary holder with a buffer tank according to the present invention. Embodiment 1 shows an example in which short capillaries are held in a straight line. However, when a long capillary is used, a long capillary holder 5 shown in FIG.
Use 0. This holder has the same structure near both ends of the capillary, an optical system for irradiating laser light, an optical system for condensing fluorescence, etc. as the holder 2 for the short capillary shown in FIG. It has a structure that can hold the capillary loop 51. In this case, it is possible to use a separation capillary having a total length of 50 cm by forming a loop having a radius of 5 cm. In order to prevent troubles such as a decrease in separation ability due to heat generation due to insufficient heat dissipation, a heat dissipation sheet 52 is interposed between overlapping portions of the capillaries.

[Embodiment 3] The present embodiment is an example of a multi-capillary having a plurality of electrophoresis paths for a fluid separation medium. Generally, as the number of capillaries increases, the number of points at which fluorescence must be measured increases by the number of capillaries. In this case, a method has been reported in which fluorescence is measured while identifying a position by scanning a laser beam for excitation, or fluorescence is detected by an area sensor or a line sensor by simultaneously irradiating all migration paths. However, these detection devices are expensive and are not suitable for simple and inexpensive devices. The method presented here is a method in which a capillary portion to be irradiated with laser light is held in a planar shape, and a laser beam is applied from the side to all the irradiated portions of all capillaries simultaneously. Although a part of the present inventor has already reported a capillary array device using this side irradiation method, in that case, an area sensor was used as a detector for detecting position separation. In the present invention, fluorescence detection is performed using the inexpensive photomultiplier tube described above. When a plurality of photomultiplier tubes are used, the production cost increases and it is difficult to produce an inexpensive device.
Therefore, an example using one photomultiplier tube and a rotating filter will be disclosed. The overall configuration of the electrophoresis apparatus according to the present embodiment is almost the same as the previous example, but the detection unit is devised. That is, a method of forming images of a plurality of light emitting points, combining filters and slits, and detecting signals from the respective light emitting points in a time-dependent manner is adopted.

FIG. 6 is a schematic diagram of the electrophoresis apparatus according to the present embodiment. Here, a case in which the number of capillaries is three will be described, but the same is true even when the number of capillaries is further increased. In this embodiment, a capillary holder 2 with a buffer tank equipped with three capillaries 1 for performing electrophoresis and a laser 3 are arranged on an upper portion of a base 4, and four kinds of three capillaries, that is, a total of 12 pieces are provided. Color separation filter 5
The filter-equipped rotating plate 6 and the photomultiplier tube 7 are disposed below the base 4. The laser light emitted from the laser 3 is reflected by the reflection mirror 8 and condensed by the irradiation lens 9 to simultaneously irradiate all three capillaries 1. Further, the fluorescent light generated from the inside of the capillary is condensed by the condensing lens 10 and disposed on the filter-equipped rotating plate 6 as three separated images by the coupling lens 13 via the excitation light removing filter 11 and the reflecting mirror 12. An image is formed on or near the color separation filter 5 and is incident on the photomultiplier 7. A fixed slit 61 is disposed in front of the filter-equipped rotating plate 6 to block excess light.

The light passing through the fixed slit 61 passes through the color separation filter 5 and is subjected to fluorescence wavelength selection. The color separation filter 5 is provided according to the number of capillaries as shown in FIG.
Are divided as shown. In the case of the illustrated example, since the number of capillaries is three, one type of color separation filters 5-1a, 5-1b, and 5-1c are shifted in the circumferential direction and have different distances from the center of the rotating shaft 71. It is located in two positions. Similarly, the other three types of color separation filters 5
-2a, 5-2b, 5-2c, and 5-3a, 5-3
b, 5-3c and 5-4a, 5-4b, 5-4c are also shifted in the circumferential direction and are arranged at three different positions from the center of the rotating shaft 71, respectively. The distance from the center of the rotation axis 71 of the imaging position 72 of the fluorescent image from the three capillaries is the same as the distance from the center of the rotation axis 71 of each color separation filter 5. As a result, the fluorescence images from the three capillaries can be substantially separated by time-sharing by the rotating plate with filter 6, and can be detected using one photomultiplier tube 7.

The four types of color separation filters 5-1, 5-1
5-2, 5-3, and 5-4 are distinguished by the position detecting cuts 15-1, 15- provided in the rotating plate 6 with the filter.
2, 15-3, 15-4, and 16 are detected by the filter position detectors 17 and 18, respectively. In the fluorescence detection, a voltage is applied to the photomultiplier tube 7 using a high-voltage power supply 24 for the photomultiplier tube, and the detected fluorescence is converted into a voltage and extracted.
The computer 25 controls the high-voltage power supply 21 for electrophoresis and the high-voltage power supply 24 for photomultiplier tubes, processes signals from the filter position detectors 17 and 18, and processes signals from the photomultiplier tube 7. Since the color separation filter 5 is provided with four kinds of different transmission wavelength bands corresponding to the phosphors to be used, D labeled with four kinds of phosphors is used.
Samples such as NA or protein can be analyzed simultaneously with three capillaries. Here, an example of three capillaries is shown, but the present invention can be applied to a system having more capillary separation units.

[Embodiment 4] Next, an embodiment in which the injection of a fluid separation medium is simplified will be described. In conventional capillary gel electrophoresis using a fluid separation medium such as a fluid polymer or a fluid gel, the connection between the capillary and the fluid separation medium injection side is switched to inject the fluid separation medium and perform electrophoresis. Was. That is,
At the time of pressurizing the fluid separation medium into the capillary, the fluid separation medium injection part and the capillary were connected, and the fluid separation medium was pressure-injected with the capillary separated from the electrophoresis buffer solution. Further, at the time of separation measurement, the capillary was shut off from the fluid separation medium injection portion, and the electrophoresis was carried out by applying a current for electrophoresis so that the end of the capillary was in contact with the buffer solution for electrophoresis. This was performed by switching the flow path switching valve, sealing the space between the fluid separation medium injection part and the capillary, and providing a gap as described above. is necessary. However, high withstand voltage switching components are expensive. Here, an inexpensive and simple method capable of injecting a fluid separation medium and supplying an electrophoretic voltage without using a flow path switching valve is disclosed.

When the fluid separation medium is injected, pressurization is required, so that the space between the injection section of the fluid separation medium and the capillary must be sealed. As a result, the capillary terminal and the buffer solution for electrophoresis are electrically insulated. On the other hand, in order to perform electrophoresis, it is necessary that the end of the capillary be in electrical contact with the buffer solution for electrophoresis and be in an energized state. In order to satisfy both of these requirements, a porous pipe is provided between the capillary and the fluid separation medium inlet in the present embodiment.

FIG. 8 shows an outline of an example of the fluid separation medium injection section of the present embodiment. The end of the capillary 1 installed in the capillary holder 2 with a buffer tank is fixed to the center of the capillary fixing seal pin 31. The fluid separation medium is connected to the connector 34 with the fluid separation medium injection nozzle and the capillary 1 in the buffer tank 20.
Is injected into the capillary 1 through the connector 34 by press-fitting. At the tip of the connector 34 with a fluid separation medium injection nozzle, a fluid separation medium injection section connector 80 made of porous material is attached.
It is in close contact with the capillary fixing seal pin 31 that fixes the capillary 1 so that a fluid separation medium can be introduced into the capillary 1.

The connector 80 for the injection part of the fluid separation medium made of porous material is in contact with the buffer solution in the buffer tank 20 for electrophoresis, and the ions in the buffer solution can pass through the porous wall. Therefore, by applying a high voltage for electrophoresis to the electrode 23 in the buffer tank 20, a voltage is applied to both ends of the capillary 1 to enable electrophoresis. On the other hand, when the fluid separation medium is injected under pressure, the porous fluid separation medium injection connector 80 has a large resistance to the passage and outflow of the fluid separation medium through the porous wall. In addition, the fluid separation medium hardly leaks outside, and the fluid separation medium can be injected into the capillary 1 without any trouble. After the fluid separation medium is injected into the capillary 1, a sample is injected from the other end of the capillary 1 according to a usual procedure, and electrophoresis separation and detection are performed by applying a migration voltage from the electrodes 22 and 23. Accordingly, a predetermined electrophoresis operation can be easily performed without having a function such as a switching valve.

[0036]

As described above, according to the present invention, a small laser light source, a detector, a capillary separation unit and the like are fixed on one substrate, and a detection unit combining a photomultiplier tube and a small rotary filter is provided. By using the same and using a simplified capillary holder and a fluid separation medium injection device, a small and inexpensive DNA analyzer can be realized. The volume of this device is about 3% of the conventional capillary gel electrophoresis device, and the price is a fraction to one tenth. Such a handy device can be used in a laboratory or a clinical site, and can be widely used in an educational site or the like.

[Brief description of the drawings]

FIG. 1 is a partially cutaway perspective view showing an example of an electrophoresis apparatus according to the present invention.

FIG. 2 is a schematic cross-sectional view of an example of an apparatus equipped with a fluid separation medium injection section and a sample injection section.

FIG. 3 is an explanatory diagram of an example of a fluid separation medium injection section.

FIG. 4 is a detailed sectional view of a sample injection section.

FIG. 5 is a conceptual diagram of a device in which a long capillary is mounted in a loop shape.

FIG. 6 is a conceptual diagram of an apparatus equipped with a plurality of capillaries.

FIG. 7 is a diagram illustrating an example of a color separation filter that enables color separation and capillary identification.

FIG. 8 is a conceptual diagram of an injection part of an apparatus for injecting a fluid separation medium through a porous pipe.

[Description of Signs] 1 ... Capillary, 2 ... Capillary holder with buffer tank, 3 ... Laser, 4 ... Base, 5; 5-1a, 5
-1b, 5-1c; 5-2a, 5-2b, 5-2c; 5
-3a, 5-3b, 5-3c; 5-4a, 5-4b, 5
-4c: color separation filter, 6: rotating plate with filter, 7: photomultiplier tube, 8: reflection mirror, 9: irradiation lens, 10: condenser lens, 11: excitation light removal filter,
12: Reflecting mirror, 13: Coupling lens, 14: Rotating motor, 15, 16: Cut for position detection, 17, 18: Filter position detector, 19, 20: Buffer tank, 21
... high-voltage power supply, 22, 23 ... electrode, 24 ... high-voltage power supply for photomultiplier tube, 25 ... computer, 31, 32 ... seal pin for fixing capillary, 33 ... injector, 34 ... connector with fluid separation medium injection nozzle, 35 ... sample cartridge holder, 36 ... sample holder, 37 ... sample injection tool, 38 ... sample holder cartridge for continuous sample injection, 39 ... sample holder, 50 ... long capillary holder, 51 ... capillary loop, 52 ... Heat radiating sheet, 61: fixed slit, 71: rotating shaft, 72: imaging position of the fluorescent image from the capillary, 80: connector for injection of fluid separation medium made of porous material

────────────────────────────────────────────────── ─── Continuation of the front page (56) References JP-A-5-93711 (JP, A) JP-A-11-502618 (JP, A) (58) Fields studied (Int. Cl. ⁷ , DB name) G01N 27/447 G01N 21/64

Claims (4)

(57) [Claims] 1. A fluorescence detector comprising: a fluorescence excitation light source; a rotary filter in which a plurality of types of optical filters are separately arranged in a circumferential direction and a radial direction; and a photodetector for detecting light passing through the optical filter. And a main body having a capillary holder mounting portion, a plurality of separation capillaries held in a horizontal direction, and a pair of buffer tanks for storing a buffer solution into which the ends of the capillaries are immersed. A capillary holder which is detachable from a capillary holder mounting portion of the main body while holding the capillary of the book, wherein excitation from the fluorescence excitation light source is performed with the capillary holder mounted on the capillary holder mounting portion of the main body. Simultaneously irradiating the plurality of capillaries with light to detect the fluorescence In, the fluorescence generated from the sample in the plurality of capillaries is separated for each capillary at different radial positions of the rotating filter and imaged, and the fluorescence generated from each capillary is separated in the circumferential direction. A capillary electrophoresis device wherein wavelengths are separated by a plurality of optical filters and detected. 2. The method of claim 1 Symbol placement capillary electrophoresis device, one end of the capillary, the installed in the buffer tank is connected to the flowable separation medium injection means has a wall portion of the porous And the porous wall of the tube member allows ions to pass therethrough but exhibits high resistance to the passage of the flowable separation medium, and the flowable medium is injected through the tube member. A capillary electrophoresis apparatus having a function of injecting the fluid separation medium from the means into the capillary. 3. A capillary electrophoresis apparatus according to claim 1 Symbol mounting, one end of the capillary is fixed in the capillary holder by the jig, the buffer tank to put buffer solution immersing one end of the capillary flowable A movable pipe member is provided in the direction of the jig, which has a connection portion with the separation medium injection means, and flows into the capillary by bringing the tip of the pipe member into close contact with the jig so as to surround one end of the capillary. A capillary electrophoresis device having a function of injecting a pressure-separated medium into a medium. 4. A capillary electrophoresis apparatus of any one of claims 1 to 3, and the sample holding member which is arranged in the buffer tank by a recess holding the sample solution to face the ends of the capillary, the sample Moving means for moving a holding member toward the end of the capillary, and having a function of bringing a sample solution into contact with the end of the capillary and injecting the sample liquid into the capillary by an electric field.