JP6178161B2 - Flow cell fixing device and nucleic acid analyzer - Google Patents

Description

  The present invention relates to a nucleic acid analyzer used for analysis of nucleic acid (for example, DNA (Deoxyribonucleic acid) and RNA (ribonucleic acid)) sequences and a flow cell fixing device mounted on the apparatus.

  Recent nucleic acid analyzers detect a large number of DNA fragments immobilized on a substrate in parallel to improve analysis throughput. In the present specification, a substrate on which a DNA fragment is immobilized is called a “flow cell”. The nucleic acid analyzer adjusts the temperature by injecting a reagent into the flow cell installed at a predetermined position in the device, and further detects the fluorescence emitted from the DNA fragment by irradiating the flow cell with excitation light. Read DNA sequence based on

  A general nucleic acid analyzer includes a holder mechanism for attaching a flow cell, a temperature adjusting mechanism for reacting a sample in the flow cell with a reagent, a liquid feeding mechanism for injecting a reagent into the flow cell, and an optical for performing irradiation / fluorescence detection. The system is composed of a stage mechanism for moving the flow cell.

  Currently, there is a need for miniaturization of nucleic acid analyzers. When downsizing the apparatus, the flow cell mounting structure is important. An example of a flow cell mounting structure is shown in Patent Document 1. Patent Document 1 shows a structure in which a plurality of flow cells are attached to a heat plate that is attached in parallel and can be adjusted in temperature.

Special table 2010-539987 gazette

  However, as in the technique described in Patent Document 1, the method of arranging a plurality of flow cells in parallel increases the size in the arrangement direction as the number of flow cells arranged in parallel increases, thereby reducing the size of the nucleic acid analyzer. Is unsuitable.

  Moreover, it is necessary to always irradiate the excitation light necessary for fluorescence detection from the vertical direction with respect to the flow cell. For this reason, when the mounting angle of the flow cell is fixed, the arrangement position of the irradiation / detection unit that irradiates the excitation light is naturally limited to the position directly above the flow cell. This restriction on the arrangement hinders miniaturization of the nucleic acid analyzer.

  Therefore, the present specification provides a flow cell fixing device that has a high degree of freedom in the mounting angle of the flow cell and can arrange a plurality of flow cells in a space-saving manner. The present specification also provides a nucleic acid analyzer miniaturized by mounting the flow cell fixing device.

  The present specification includes a plurality of means for solving the above-described problems. A flow cell fixing device as one form thereof includes a holder unit that adjusts the temperature of a flow cell that holds a sample on a substrate, and a rotation that extends in a horizontal direction. A rotary drum that rotates in a vertical direction about an axis, a rotary drum having a plurality of attachment portions of the holder unit on a rotary surface thereof, a rotary mechanism that rotationally drives the rotary drum, and the rotary drum And a sensor for detecting information related to rotation of the motor.

  Another embodiment of the flow cell fixing device is a holder unit that adjusts the temperature of a flow cell that holds a sample on a substrate, and a rotary drum that rotates in a horizontal direction around a rotation axis extending in a vertical direction. A rotary drum having a plurality of attachment portions of the holder unit on the rotation surface; a rotation mechanism that rotationally drives the rotary drum; and a sensor that detects information related to rotation of the rotary drum.

  According to the present invention, the space-saving structure of the flow cell can be realized, and the nucleic acid analyzer can be further reduced in size as compared with the conventional one. Problems, configurations, and effects other than those described above will become apparent from the following description of embodiments.

1 is a diagram illustrating a configuration example of a nucleic acid analyzer according to Example 1. FIG. FIG. 3 is a perspective view for explaining an external configuration example of the flow cell fixing device according to the first embodiment. FIG. 3 is a perspective view illustrating an example of an exploded configuration of the flow cell fixing device according to the first embodiment. FIG. 6 is a perspective view for explaining an external configuration example of a flow cell fixing device according to a second embodiment. FIG. 6 is a perspective view illustrating an example of an exploded configuration of a flow cell fixing device according to a second embodiment.

  Hereinafter, embodiments of the present invention will be described with reference to the drawings. The embodiment of the present invention is not limited to the examples described later, and various modifications are possible within the scope of the technical idea.

[Example 1]
(Overall configuration of nucleic acid analyzer)
FIG. 1 shows a schematic configuration of a nucleic acid analyzer 1 according to an embodiment. The nucleic acid analyzer 1 contains the components shown in FIG. 1 in its housing. The nucleic acid analyzer 1 has constituent elements in addition to the constituent elements shown in FIG.

  The flow cell 2 is formed with an inlet, an outlet, a reaction region, and a flow path connecting them. Although the structure of the flow cell 2 is various, at least a region portion for detecting fluorescence by irradiating excitation light is made of a material that is transparent to the wavelength band of excitation light or fluorescence.

  The flow cell 2 is detachably attached to an attachment surface of a holder unit 3 with a temperature adjustment mechanism (hereinafter referred to as “holder unit 3”). For example, the flow cell 2 is attached to the attachment surface of the holder unit 3 by a vacuum chuck method. For example, the flow cell 2 can also be attached to the attachment surface of the holder unit 3 using a fastener or the like.

  The holder unit 3 is attached to a cylindrical rotary drum 4. In this embodiment, the holder unit 3 is detachably attached to the rotary drum 4. However, the holder unit 3 and the rotary drum 4 may have an integral structure. The rotary drum 4 of this embodiment is rotationally driven in the vertical direction around a rotation shaft (not shown) extending in the horizontal direction.

  The rotary drum 4 is rotationally driven by a rotation mechanism (not shown). The rotation mechanism rotationally drives the rotary drum 4 by a rotational force transmitted through a belt, a gear, a cam or the like. Four holder units 3 are attached to the side surface (rotation surface) of the rotary drum 4 at intervals of 90 ° along the circumferential direction. The rotating shaft of the rotary drum 4 is supported by a U-shaped (U-shaped) drum holder 4A.

  The holder unit 3 is attached to an attachment surface provided on the rotary drum 4 by, for example, a vacuum chuck method. Moreover, the holder unit 3 can also be attached to the attachment surface provided in the rotary drum 4 using a fastener. Detailed structures of the holder unit 3 and the rotary drum 4 will be described later.

  The drum holder 4A is mounted on a stage unit 5A that slides in the X-axis direction. The stage unit 5A is mounted on a stage unit 5B that slides in the Y-axis direction. The reagent container 6 contains a reagent and washing water. The liquid feeding unit 7 is a drive source that sucks the reagent contained in the reagent container 6 through the pipe 6 </ b> A and injects it into the flow cell 2. In the present specification, the liquid feeding unit 7 is also referred to as a reagent injection unit. The tube 6A has elasticity, for example, and can follow the movement of the stage units 5A and 5B. The switching valve 10 is located in the middle of the pipe 6A and connects the liquid feeding unit 7 and the reagent container 6 to form a flow path, or connects the liquid feeding unit 7 and the flow cell 2 to form a flow path. Execute the switch.

  The irradiation / detection unit 8 includes a light source that emits excitation light toward a sample fixed to the flow cell 2 and a sensor that detects fluorescence emitted by the sample that has been irradiated with excitation light. The irradiation / detection unit 8 is arranged at an arbitrary position perpendicular to the rotation surface of the rotary drum 4. Since the flow cell 2 moves as the rotary drum 4 rotates, the excitation light can enter the flow cell 2 perpendicularly as long as it is perpendicular to the rotation surface of the rotary drum 4. However, in this embodiment, the irradiation / detection unit 8 is mounted above the rotary drum 4.

  The waste liquid container 9 is a container for storing the waste liquid from the flow cell 2. The waste liquid in the flow cell 2 is discarded to the waste liquid container 9 through the flow path of the holder unit 3, the flow path in the rotary drum 4, and the pipe 6A in order. In addition, an analysis unit 11 and a display unit 12 are arranged in the housing of the nucleic acid analyzer 1. The analysis unit 11 analyzes the fluorescence detected by the irradiation / detection unit 8 and detects the base sequence of the DNA contained in the sample. The display unit 12 displays the analysis result of the analysis unit 11.

  The analysis unit 11 according to the present embodiment is configured so that the operation of the rotary drum 4, the operations of the stage units 5A and 5B, the operation of the liquid feeding unit 7, the operation of the irradiation / detection unit 8, the switching valve according to the progress of the analysis process. 10 operations and the operation of the display unit 12 are controlled. That is, the analysis unit 11 functions as a control unit.

(Observation operation)
Next, an outline of operations executed in the nucleic acid analyzer 1 will be described. First, in the pretreatment apparatus, the flow cell 2 on which many DNA fragments having a diameter of about 1 micrometer are fixed is attached to the holder unit 3. Here, it is assumed that only one flow cell 2 is attached to the rotary drum 4. The flow cell 2 is attached to the holder unit 3 located at the highest point (position facing the irradiation / detection unit 8) among the holder units 3 that rotate together with the rotary drum 4.

  Next, the switching valve 10 closes the valve of the tube 6A connected to the rotary drum 4 side, and opens the valve of the tube 6A connected to the sample container 6 side. Thereby, a flow path is formed between the liquid feeding unit 7 and the reagent container 6. Thereafter, the liquid feeding unit 7 sucks the reagent from the reagent container 6 through the tube 6A. In the case of the present embodiment, the pipe 6 </ b> A is connected to the flow path in the drum through the inside of the rotation shaft of the rotary drum 4.

  When the reagent suction is completed, the switching valve 10 closes the valve of the tube 6A connected to the sample container 6 side, and opens the valve of the tube 6A connected to the rotary drum 4 side. Thereby, a flow path is formed between the liquid feeding unit 7 and the rotary drum 4. Thereafter, the liquid feeding unit 7 injects the reagent into the rotary drum 4 through the tube 6A.

  The reagent injected into the rotary drum 4 is injected into the flow cell 2 through the internal flow path of the rotary drum 4 and the internal flow path of the holder unit 3. For example, a valve (not shown) that is controlled by the analysis unit 11 is provided inside the rotary drum 4 so that the reagent is injected only into the specific flow cell 2.

  After injection of the reagent, the reaction between the DNA fragment and the reagent occurs in the reaction region of the flow cell 2 by adjusting the temperature by the holder unit 3. In order to observe the reacted DNA fragment, the stage units 5A and 5B slide in the XY plane. When the flow cell 2 is positioned at the observation position, the irradiation / detection unit 8 irradiates the reaction region with excitation light. Further, the irradiation / detection unit 8 detects fluorescence emitted from a plurality of DNA fragments existing in the reaction region by the light receiving unit.

  When the fluorescence detection is completed for one observation region, the stage units 5A and 5B move the drum holder 4A minutely in the horizontal plane, and the next observation region of the flow cell 2 is positioned immediately below the irradiation / detection unit 8. Thereafter, irradiation of excitation light to the observation region and detection of fluorescence are performed by the same method as described above. This observation operation is repeatedly performed a plurality of times. When the observation is completed for all the observation regions, the liquid feeding unit 7 sucks the washing water stored in the reagent container 6 and then injects it into the flow cell 2. The flow path in the flow cell 2 is washed by injecting the washing water. Thereafter, another reagent is injected into the flow cell 2, and after the above-described observation, a series of operations for further washing is repeated a plurality of times. After execution of the observation operation, the analysis unit 11 reads the DNA sequence of the DNA fragment.

  When a plurality of flow cells 2 are attached to the rotary drum 4, when the observation of one flow cell 2 is completed, the rotary drum 4 is rotated around the rotation axis, and another flow cell 2 is irradiated and detected. It moves directly under the unit 8 and starts observation for the observation position. Thereafter, the same observation operation is performed for another flow cell 2.

(Configuration of flow cell fixing device)
A configuration example of the flow cell fixing device 20 mounted on the nucleic acid analyzer 1 will be described with reference to FIGS. 2 and 3. 2 shows an external configuration of the flow cell fixing device 20, and FIG. 3 shows an exploded configuration of the flow cell fixing device 20.

  The flow cell fixing device 20 includes a flow cell 2, a holder unit 3, a rotary drum 4, a drum holder 4A, and a rotation mechanism (not shown). Up to four holder units 3 in the present embodiment can be installed on the rotary drum 4 that rotates in the vertical direction. For this reason, on the side surface (rotating surface) of the cylindrical rotary drum 4, four concave accommodating portions 4 </ b> B are formed. In the present embodiment, the accommodating portion 4B has a rectangular cross section. The four accommodating portions 4B are arranged at 90 ° intervals in the circumferential direction.

  In the case of the present embodiment, the accommodating portion 4B reaches from the one end surface of the cylindrical rotary drum 4 to the other end surface. The depth (inner surface height) of the accommodating portion 4B is formed so as not to hinder the rotation of the rotary drum 4 in a state where the flow cell 2 is attached to the holder unit 3. That is, the accommodating part 4B is formed so that the whole of the flow cell 2 and the holder unit 3 can be accommodated.

  By attaching a plurality of flow cells 2 on the rotating surface as in this embodiment, the dimension required in the horizontal direction can be greatly reduced compared to the case where the same number of flow cells 2 are attached in the parallel direction. it can. In the case of the present embodiment, the flow cell 2 is attached to the rotating surface of the rotary drum 4 that is rotationally driven, so that the installation angle of the flow cell 2 is widened. As a result, the irradiation / detection unit 8 can be arranged at an arbitrary position facing the rotation surface. This means that the degree of freedom of the arrangement position of the irradiation / detection unit 8 in the casing of the nucleic acid analyzer 1 can be increased. As a result, the internal volume of the nucleic acid analyzer 1 can be further reduced as compared with the conventional apparatus.

  The holder unit 3 includes a heat plate 21 that can be adjusted in temperature, a Peltier element (not shown) that adjusts the temperature of the heat plate 21, a heat sink 22 for heat dissipation, and a holder base 23. In the case of the present embodiment, the heat plate 21 and the holder base 23 (also the heat sink 22 if necessary) are formed with a flow path penetrating the inside, and the flow of the solution into and out of the flow cell 2 through the flow path. Is done.

  Here, the opening 24 exposed on the surface of the heat plate 21 is disposed at a position facing the inlet and outlet of the flow cell 2. Similarly, the opening 24 exposed on the bottom surface of the holder base 23 is disposed at a position facing the opening 24 exposed on the bottom surface of the housing portion 4B of the rotary drum 4. A mechanism or mechanism for preventing liquid leakage is employed between the attachment surface of the flow cell 2 and the heat plate 21 and the attachment surface of the holder base 23 and the rotary drum 4. For example, a structure in which the periphery of the opening 24 is surrounded by a sealing ring (for example, rubber) or a mechanism in which the flow cell 2 and the mounting surface of the heat plate 21 are closely attached by a vacuum chuck is employed.

  When the vacuum chuck method is adopted for mounting the flow cell 2 and the holder base 23, an exhaust passage is formed inside the rotary drum 4, and vacuum is passed through the passage formed inside the rotary shaft of the rotary drum 4. Allow connection to the pump. The drum holder 4A is provided with a sensor 25 that detects the rotational direction, rotational position, and rotational speed of the rotary drum 4. Information detected by the sensor 25 is given to the analysis unit 11.

(Effects of Example 1)
As described in the present embodiment, by using the flow cell fixing device 20 that can mount a plurality of flow cells 2 on the rotating surface of the rotary drum 4, a space necessary for mounting a plurality of flow cells can be reduced as compared with the conventional case. Can be greatly reduced.

  Further, since the installation angle of the flow cell 2 is widened (because it is widened to 360 °), the irradiation / detection unit 8 can be disposed at an arbitrary position facing the rotation surface of the rotary drum 4. As a result, the internal volume of the nucleic acid analyzer 1 can be further reduced as compared with the conventional apparatus.

[Example 2]
(Overall configuration of nucleic acid analyzer)
The constituent elements constituting the nucleic acid analyzer 1 are basically the same as those in the first embodiment. However, in the case of the present embodiment, unlike the first embodiment, the flow cell fixing device 30 having the rotary drum 4 that rotates in the horizontal direction around the rotation axis extending in the vertical direction is mounted. The observation operation of this example is the same as that of Example 1.

(Configuration of flow cell fixing device)
A configuration example of the flow cell fixing device 30 mounted on the nucleic acid analyzer 1 will be described with reference to FIGS. 4 and 5. 4 shows an external configuration of the flow cell fixing device 30, and FIG. 5 shows an exploded configuration of the flow cell fixing device 30.

  The flow cell fixing device 30 includes a flow cell 2, a holder unit 3, a rotary drum 4, a drum holder 4A, and a rotation mechanism (not shown). However, as described above, the rotary drum 4 in the present embodiment is rotationally driven in the lateral direction around the rotational axis extending in the vertical direction.

(Effects of Example 2)
Also in this embodiment, since a plurality of flow cells 2 can be mounted on the rotation surface, the dimension required in the horizontal direction is greatly reduced as compared with the case where the same number of flow cells 2 are mounted in the parallel direction. be able to.

  Moreover, since the flow cell 2 is attached to the rotating surface of the rotary drum 4 that is rotationally driven, the arrangement angle of the flow cell 2 is widened. As a result, the irradiation / detection unit 8 can be arranged at an arbitrary position facing the rotation surface. As a result, the degree of freedom of the arrangement position of the irradiation / detection unit 8 in the casing of the nucleic acid analyzer 1 is increased, and the internal volume of the nucleic acid analyzer 1 can be further reduced as compared with the conventional apparatus.

[Other embodiments]
The present invention is not limited to the configuration of the embodiment described above, and includes various modifications. For example, the number of accommodating portions 4B may be two, three, or five or more. In addition, for example, the nucleic acid analyzer 1 may include a plurality of the flow cell fixing devices 20 or 30 described above. In addition, in order to demonstrate this invention in an easy-to-understand manner, the above-described embodiments are described in detail for some embodiments, and it is not always necessary to have all the configurations described. Further, a part of a certain embodiment can be replaced with the configuration of another embodiment, and the configuration of another embodiment can be added to the configuration of a certain embodiment. It is also possible to add other configurations to the configuration of each embodiment, replace a partial configuration of each embodiment with another configuration, or delete a partial configuration of each embodiment.

DESCRIPTION OF SYMBOLS 1 ... Nucleic acid analyzer 2 ... Flow cell 3 ... Holder unit 4 with temperature control mechanism ... Rotary drum 4A ... Drum holder 4B ... Storage part 5A, 5B ... Stage unit 6 ... Reagent container 6A ... Tube 7 ... Liquid feeding unit 8 ... Irradiation -Detection unit 9 ... Waste liquid container 10 ... Switching valve 11 ... Analysis unit (control unit)
DESCRIPTION OF SYMBOLS 12 ... Display part 20 ... Flow cell fixing device 21 ... Heat plate 22 ... Heat sink 23 ... Holder base 24 ... Opening 25 ... Sensor 30 ... Flow cell fixing device

Claims (14)

A holder unit for adjusting the temperature of the flow cell substrate holding the specimen thereon,
A rotary drum that rotates in the vertical direction around a rotation axis extending in the horizontal direction, and a rotary drum having a plurality of attachment portions of the holder unit on the rotation surface;
A rotation mechanism for rotating the rotary drum;
Have a sensor for detecting information about the rotation of the rotary drum,
The attachment portion is a flow cell fixing device that arranges and fixes the flow cell substrate on the rotation surface . In the flow cell fixing device according to claim 1,
The attachment portion is a concave storage portion that stores the holder unit. In the flow cell fixing device according to claim 2,
The flow cell fixing device according to claim 1, further comprising: an opening of a rotary drum side channel connected to the holder unit side channel on an inner surface of the housing unit. A holder unit for adjusting the temperature of the flow cell substrate holding the specimen thereon,
A rotary drum that rotates in a horizontal direction around a rotation axis extending in a vertical direction, and a rotary drum having a plurality of attachment portions of the holder unit on the rotation surface;
A rotation mechanism for rotating the rotary drum;
Have a sensor for detecting information about the rotation of the rotary drum,
The attachment portion is a flow cell fixing device that arranges and fixes the flow cell substrate on the rotation surface . In the flow cell fixing device according to claim 4,
The attachment portion is a concave storage portion that stores the holder unit. In the flow cell fixing device according to claim 5,
The flow cell fixing device according to claim 1, further comprising: an opening of a rotary drum side channel connected to the holder unit side channel on an inner surface of the housing unit. A flow cell substrate holding the flow path is formed specimen thereon,
A holder unit that adjusts the temperature of the flow cell substrate mounted on a support surface, and a rotary drum that rotates in the vertical direction around a rotation axis that extends in the horizontal direction, and an attachment portion of the holder unit is provided on the rotation surface. A flow cell fixing device having a plurality of rotary drums, a rotation mechanism that rotationally drives the rotary drums, and a sensor that detects information related to rotation of the rotary drums;
A reagent injection part for injecting a reagent into the flow path of the flow cell substrate ;
An irradiation unit for irradiating the flow cell substrate with excitation light;
A detection unit for detecting fluorescence from a sample in the flow cell substrate ;
An analysis unit for analyzing the fluorescence detected by the detection unit;
A display unit for displaying an analysis result by the analysis unit;
I have a,
The attachment part is a nucleic acid analyzer for disposing and fixing the flow cell substrate on the rotating surface . The nucleic acid analyzer according to claim 7,
The nucleic acid analyzer according to claim 1, wherein the attachment portion is a concave accommodating portion that accommodates the holder unit. The nucleic acid analyzer according to claim 8,
The nucleic acid analyzer according to claim 1, wherein an opening of a rotary drum side channel connected to the holder unit side channel is provided on an inner surface of the housing unit. The nucleic acid analyzer according to claim 7,
The said irradiation part and the said detection part are arrange | positioned in the position facing the rotation surface of the said rotary drum. The nucleic acid analyzer characterized by the above-mentioned. A flow cell substrate holding the specimen on the flow path of the sample is formed,
A holder unit for adjusting the temperature of the flow cell substrate mounted on a support surface, and a rotary drum that rotates in a lateral direction around a rotation axis extending in a vertical direction, and an attachment portion of the holder unit is provided on the rotation surface. A flow cell fixing device having a plurality of rotary drums, a rotation mechanism that rotationally drives the rotary drums, and a sensor that detects information related to rotation of the rotary drums;
A reagent injection part for injecting a reagent into the flow path of the flow cell substrate ;
An irradiation unit for irradiating the flow cell substrate with excitation light;
A detection unit for detecting fluorescence from a sample in the flow cell substrate ;
An analysis unit for analyzing the fluorescence detected by the detection unit;
A display unit for displaying an analysis result by the analysis unit;
I have a,
The attachment part is a nucleic acid analyzer for disposing and fixing the flow cell substrate on the rotating surface . The nucleic acid analyzer according to claim 11, wherein
The nucleic acid analyzer according to claim 1, wherein the attachment portion is a concave accommodating portion that accommodates the holder unit. The nucleic acid analyzer according to claim 12,
The nucleic acid analyzer according to claim 1, wherein an opening of a rotary drum side channel connected to the holder unit side channel is provided on an inner surface of the housing unit. The nucleic acid analyzer according to claim 11, wherein
The said irradiation part and the said detection part are arrange | positioned in the position facing the rotation surface of the said rotary drum. The nucleic acid analyzer characterized by the above-mentioned.

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