JP2009300149A - Inspection device - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an inspection device capable of correctly detecting the liquid in a microchannel even if the temperature of a microchip changes. <P>SOLUTION: The inspection device, which moves the specimen and reagent stored in the channel of the microchip to fill a detection part and measures a reaction result, has a liquid detecting means for outputting the detection signal corresponding the amount of the liquid arriving at the predetermined position of the channel, a threshold value calculating means for calculating a threshold value on the basis of the value obtained by delaying the detection signal by a predetermined time, and an arrival judging means for judging the presence of the liquid on the basis of the detection signal and the threshold value. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  The present invention relates to an inspection apparatus.

  In recent years, by making full use of micromachine technology and ultrafine processing technology, devices and means (for example, pumps, valves, flow paths, sensors, etc.) for performing conventional sample preparation, chemical analysis, chemical synthesis, etc. have been miniaturized. A system integrated on a chip has been developed (see, for example, Patent Document 1). This is also referred to as μ-TAS (Micro total Analysis System), bioreactor, lab-on-chip, biochip, and microchip. Its application is expected in the field of measurement and agricultural production. In particular, as seen in genetic testing, when complicated processes, skilled techniques, and operation of equipment are required, the cost, required sample amount, and required time can be reduced by using μ-TAS.

The present applicant encloses a reagent or the like in the microchannel of the microchip, injects a driving liquid into the microchannel by a micropump to move the liquid such as the specimen and the reagent, and flows the liquid to the reaction unit and then to the detection unit Thus, an inspection apparatus capable of measuring a reaction result with a specimen such as blood has been proposed (for example, see Patent Document 2).
JP 2004-28589 A JP 2006-149379 A

  In such an inspection apparatus, it is important to accurately control the leading position of the liquid in the fine channel. Therefore, it is desirable to detect the leading position of the liquid in the fine flow path using an optical sensor or the like and control according to the information.

  On the other hand, at the time of inspection, it is necessary to heat the microchip to a temperature suitable for the reaction between the specimen and the reagent. The sensor that detects the liquid in the fine flow path needs to be placed close to the microchip in order to increase the detection accuracy. However, if it is placed close to the microchip, the heat generated by the Peltier element or the like heats the microchip. The output of the sensor may fluctuate due to the influence of.

  In addition, microchips are often stored in a refrigerated state in order to prevent deterioration of reagents and the like stored therein. However, when the microchip is heated to a temperature suitable for the reaction during the inspection, changes occur such as causing condensation on the wall surface in the fine flow path, or disappearance of the condensation that was in the condensation state before heating. For this reason, there arises a problem that the sensor for detecting the liquid in the fine flow path cannot detect the liquid or erroneously detects it.

  The present invention has been made in view of the above problems, and an object of the present invention is to provide an inspection apparatus that can correctly detect a liquid in a fine channel even if the temperature of a microchip changes.

  The object of the present invention can be achieved by the following constitution.

1. In an inspection device for measuring the result of moving the sample and reagent stored in the flow path of the microchip and filling the detection unit and reacting them,
Liquid detection means for outputting a detection signal corresponding to the amount of liquid that has reached a predetermined position of the flow path;
Threshold calculation means for calculating a threshold based on a value obtained by delaying the detection signal for a predetermined time;
Arrival determination means for determining the presence or absence of the liquid based on the detection signal and the threshold;
An inspection apparatus comprising:

2. The threshold calculation means includes
2. The inspection apparatus according to 1, wherein a threshold value is calculated by adding or subtracting a predetermined value to a value obtained by delaying the detection signal for a predetermined time.

3. The threshold calculation means includes
2. The inspection apparatus according to claim 1, wherein a threshold value is calculated by multiplying a value obtained by delaying the detection signal by a predetermined time by a predetermined value.

  According to the present invention, the threshold value is calculated based on a value obtained by delaying the detection signal corresponding to the amount of the liquid reaching the predetermined position of the flow path for a predetermined time, and the presence / absence of the liquid is determined based on the detection signal and the threshold value. Since the determination is made, it is possible to provide an inspection apparatus that can correctly detect the liquid in the fine channel even if the temperature of the microchip changes.

  Hereinafter, embodiments of the present invention will be described with reference to the drawings.

  FIG. 1 is an external view of an inspection apparatus 80 according to an embodiment of the present invention.

  The inspection device 80 is a device that automatically detects a reaction between a sample previously injected into the microchip 1 and a reagent and displays the result on the display unit 84.

  The housing 82 of the inspection apparatus 80 has an insertion port 83, and the microchip 1 is inserted into the insertion port 83 and set inside the housing 82. The insertion port 83 is sufficiently higher than the thickness of the microchip 1 so as not to contact the insertion port 83 when the microchip 1 is inserted. Reference numeral 85 denotes a memory card slot, 86 denotes a print output port, 87 denotes an operation panel, and 88 denotes an input / output terminal.

  The person in charge of inspection inserts the microchip 1 in the direction of the arrow in FIG. 1 and operates the operation panel 87 to start the inspection. Inside the inspection device 80, the reaction in the microchip 1 is automatically inspected, and when the inspection is completed, the result is displayed on the display unit 84 constituted by a liquid crystal panel or the like. The inspection result can be output from the print output port 86 or stored in a memory card inserted into the memory card slot 85 by operating the operation panel 87. Further, data can be stored in the personal computer or the like from the external input / output terminal 88 using, for example, a LAN cable.

  The inspection person takes out the microchip 1 from the insertion port 83 after the inspection is completed.

  Next, an example of the microchip 1 according to the embodiment of the present invention will be described with reference to FIG.

  FIG. 2 is an external view of the microchip 1 in the embodiment of the present invention, and FIG. 3 is an enlarged view of the periphery of the detection unit 111.

  As shown in the side view of the microchip 1 in FIG. 2A, the microchip 1 includes a groove forming substrate 108 and a covering substrate 109 that covers the groove forming substrate 108.

  FIG. 2B is a plan view of the microchip 1, and illustrates the grooves of the groove forming substrate 108 that can be seen through the transparent coated substrate 109. A flow path is formed by covering the grooves of the groove forming substrate 108 with the covering substrate 109. The microchip 1 is provided with minute groove-like channels 250 (microchannels) and functional parts (channel elements) for performing inspections, sample processing, and the like in an appropriate manner according to the application. Has been.

  The flow path is formed on the order of micrometers, and for example, the width is several μm to several hundred μm, preferably 10 to 200 μm, and the depth is about 25 to 500 μm, preferably 25 to 250 μm.

  In the present embodiment, a microchip 1 used for a process of performing amplification and detection of a specific gene will be described as an example.

  Reference numerals 110a and 110b in FIG. 2B denote injection ports that communicate with the flow path inside the microchip 1. The driving liquid is injected from each of the injection ports 110 to drive the internal specimen, reagent, and the like. In the microchip 1 of the present embodiment, as shown in FIG. 2B, the configuration of the flow path starting from the injection port 110a is completely the same as the configuration of the flow path starting from the injection port 110b. Distinguish between calls. Each component is distinguished by adding a and b.

  Reference numerals 121a and 121b denote sample storage units for storing samples. The sample storage units 121a and 121b have deeper grooves than other flow paths in order to store a predetermined amount of sample. In the present embodiment, a description will be given assuming that samples are stored in the sample storage units 121a and 121b in advance.

  Reference numerals 120a and 120b denote reagent storage parts that store the first reagent, 122a and 122b denote reagent storage parts that store the second reagent, and 123a and 123b denote reagent storage parts that store the third reagent.

  When the driving liquid is injected from the injection ports 110a and 110b, the sample stored in the sample storage units 121a and 121b is pushed out to the flow channel 250 and is injected into the downstream reagent storage units 120a and 120b. The sample and the first reagent pushed out from the reagent storage units 120a and 120b are respectively injected into the downstream reagent storage units 122a and 122b through the flow channel 250. The specimen and the first reagent push out the second reagent from the reagent storage parts 122a and 122b.

  The sample, the first reagent, and the second reagent pass through the flow path 250 and are injected into the downstream reagent storage units 123a and 123b, respectively, to push out the third reagent.

  A mixing unit 130a, a mixing unit 130b, a mixing unit 131a, and a mixing unit 131b are provided downstream of the reagent storage units 123a and 123b. The flow channel 250aa, the sample flowing through the flow channel 250ba, the first reagent, The second reagent and the third reagent are mixed in each mixing unit.

  Liquid reservoirs 140a and 140b are provided in the flow channel 250aa and the flow channel 250ba on the upstream side near the mixing unit 130a and the mixing unit 130b. The liquid reservoirs 140a and 140b are provided to detect that the third reagent has been pushed out of the reagent storage parts 123a and 123b and the leading portion has reached the vicinity of the mixing part 130a and the mixing part 130b.

  The liquid reservoirs 140 a and 140 b are shallower than the mixing part 130 a and mixing part 130 b and deeper than the other flow paths 250. The depth of the grooves of the mixing portion 131a and the mixing portion 131b is, for example, 1.5 mm, the depth of the grooves of the liquid reservoir portions 140a, 140b is, for example, 0.6 mm, and the depth of the grooves of the other flow paths is, for example, 0.25 mm. is there.

  The sample, the first reagent, the second reagent, and the third reagent mixed in the mixing unit 130a, the mixing unit 130b and the mixing unit 131a, and the mixing unit 131b are injected into the detection units 111a and 111b. As will be described later, the microchip 1 is heated or absorbed inside the inspection apparatus 80 to cause the specimen and the reagent to react at a predetermined temperature for a predetermined time.

  The detection units 111a and 111b are provided for optically detecting the reaction between the specimen and the reagent, and have a groove deeper than the other flow paths to accommodate a predetermined amount of the specimen and the reagent.

  Liquid reservoirs 141a and 141b are provided downstream of the detection units 111a and 111b so that it can be detected that the detection units 111a and 111b are filled with a mixed solution of a reagent and a specimen. The liquid reservoirs 141a and 141b are shallower than the detectors 111a and 111b, and deeper than the other channels.

  An example of the shapes of the detection unit 111 and the liquid reservoir 141 will be described with reference to FIG. FIG. 3A is a plan view, and FIG. 3B is a cross-sectional view of the detection unit 111.

  The groove depth d3 of the detectors 111a and 111b is, for example, 1.5 mm, the groove depth d2 of the liquid reservoirs 141a and 141b is, for example, 0.6 mm, and the groove depth d1 of the other flow paths is, for example, 0.25 mm. It is. It is desirable that the liquid reservoirs 141a and 141b be provided as close as possible to the downstream of the detection units 111a and 111b so that it can be detected as soon as possible that the detection units 111a and 111b are filled. Reference numeral 19 denotes a liquid detection unit using, for example, a photo reflector, which irradiates the liquid reservoirs 141a and 141b with light to detect reflected light and outputs an electrical signal.

  When the detection units 111a and 111b emit light from the detection unit 22 (not shown in FIG. 3), the reagent that has reacted with the specimen emits, for example, fluorescence. Therefore, the detection unit 22 measures the amount of fluorescence and displays the reaction result. measure.

  Next, materials used for the groove forming substrate 108 and the covering substrate 109 constituting the microchip 1 will be described.

  The microchip 1 is desired to be excellent in processability, non-water absorption, chemical resistance, weather resistance, cost and the like. In consideration of the structure, application, detection method, etc. of the microchip 1, The material of chip 1 is selected. Various known materials can be used as the material, and usually the substrate and the flow path element are formed by appropriately combining one or more materials in accordance with individual material characteristics.

  Since the detection unit 111 optically detects a fluorescent substance or the like, at least this part of the covering substrate 109 uses a light transmitting material (for example, alkali glass, quartz glass, transparent plastics) so that light can be transmitted. It is necessary to.

  When optically detecting the liquid stored in the liquid reservoirs 140a and 140b and the liquid reservoirs 141a and 141b, at least the portion covering the liquid reservoirs 140a and 140b and the liquid reservoirs 141a and 141b of the coated substrate 109 is It is necessary to configure with a transparent member such as glass or resin. In addition, when detecting a liquid using an ultrasonic wave, an electromagnetic wave, etc., it is not necessary to use a transparent member.

  FIG. 4 is a perspective view for explaining an example of the internal configuration of the inspection apparatus 80 in the embodiment of the present invention, and FIG. 5 is a cross-sectional view showing an example of the internal configuration of the inspection apparatus 80 in the embodiment of the present invention. is there. FIG. 6 is an explanatory diagram illustrating a state where the detection unit 111 is filled with liquid. Each part will be described with reference to the coordinate axes of X, Y, and Z shown in FIGS.

  The inspection device 80 includes a temperature adjustment unit 152, a temperature adjustment unit 153, a detection unit 22, a micro pump unit 75, a packing 90, a driving liquid tank 91, a feed screw 301, a joint 302, a detection unit drive motor 61, and the like. 4 and 5 show a state in which the microchip 1 is in close contact with the packing 90b.

  Hereinafter, an example of the internal configuration of the inspection apparatus 80 will be described with reference to FIGS. 4, 5, and 6.

  The temperature adjustment unit 152 is driven by a drive member (not shown) and is movable in the Z-axis direction. In the initial state, the temperature adjustment unit 152 is raised from the state of FIG. 4 by the thickness of the microchip 1 by the driving member. Then, the microchip 1 can be inserted and removed in the Y-axis direction, and the person inspecting inserts the microchip 1 from the insertion port 83 until it comes into contact with a regulating member (not shown). When the microchip 1 is inserted to a predetermined position, the chip detection unit 95 using a photo interrupter or the like detects the microchip 1 and is turned on.

  The temperature adjustment unit 152 and the temperature adjustment unit 153 incorporate a Peltier element, a heater, a power supply device, a temperature control device, and the like, and adjust the surface of the microchip 1 to a predetermined temperature by generating heat or absorbing heat. This unit promotes the reaction of the filled liquid.

  The temperature adjustment unit 152 and the microchip 1 are lowered by the driving member, and the microchip 1 is brought into close contact with the temperature adjustment unit 152, the temperature adjustment unit 153, and the packing 90b.

  In the present embodiment, as shown in FIG. 5, both surfaces of the microchip 1 are sandwiched between the temperature adjustment unit 152 and the temperature adjustment unit 153 and adjusted to a predetermined temperature.

  As shown in the cross-sectional view of FIG. 5, the liquid detection unit 20 and the liquid detection unit 19 are disposed on the substrate 21 of the temperature adjustment unit 153. 5 shows a cross section along the liquid reservoir 140 and the liquid reservoir 141 of the a channel or b channel, both channels have the same configuration, and will be described without particular distinction in the following description.

  The liquid detection unit 20 and the liquid detection unit 19 are, for example, a photo reflector including a light emitting unit and a light receiving unit, and optically store the liquid stored in the liquid storage unit 140 and the liquid storage unit 141 on the detection substrate 21. It is mounted at each position where it can be detected. On the surface of the temperature adjustment unit 153 in contact with the microchip 1, openings are provided in portions corresponding to the optical paths of the liquid detection unit 20 and the liquid detection unit 19 as shown in FIG. Further, the temperature adjustment unit 153 and the substrate 21 are provided with through holes at corresponding positions so that the detection unit 111 can irradiate the detection unit 111 with excitation light and receive fluorescence.

  Next, an example of a method for optically detecting the liquid stored in the liquid reservoirs 140a and 140b and the liquid reservoirs 141a and 141b of the microchip 1 will be described.

  The liquid detection unit 20 irradiates the liquid reservoir 140 with light, receives the light reflected by the liquid reservoir 140, and outputs a detection signal corresponding to the amount of liquid based on a change in light amount and color. Similarly, the liquid detector 19 receives the light reflected by the liquid reservoir 141 and outputs a detection signal corresponding to the amount of liquid. The liquid detection part 19 and the liquid detection part 20 are the liquid detection means of this invention.

  A detection signal output from the liquid detection unit 19 will be described with reference to FIG.

  6A shows a state where the detection unit 111 is not sufficiently filled with the liquid 57, and FIG. 6B shows a state where the detection unit 111 is filled with the liquid 57. When the leading portion of the liquid 57 reaches the downstream side of the detection unit 111 as shown in FIG. 6B, the liquid 57 is also filled with the liquid 57. Then, the reflected light from the liquid reservoir 141 suddenly decreases, and the detection signal output from the liquid detector 19 also shows a sharp change. By detecting a change in the detection signal as will be described in detail later, it can be known that the detection unit 111 is filled with the liquid 57. The detection signal output from the liquid detection unit 20 when the liquid reservoir 140 is filled with the liquid 57 similarly shows a steep change.

  In this embodiment, an example will be described in which the liquid reservoirs 140a and 140b and the liquid reservoirs 141a and 141b are irradiated with light and reflected light is detected. However, the light transmitted through the liquid reservoir 140 and the liquid reservoir 141 is described. The present invention can also be applied to the case where a detection signal corresponding to the amount of liquid is output by detecting an ultrasonic wave or radio wave.

  Returning to FIG. 4, each part will be described.

  The detection unit 22 includes a light emitting unit and a light receiving unit, and is configured to optically separate the fluorescence emitted from the reagent that has reacted with the specimen by irradiating the detection unit 111 with light and to receive the light in the light receiving unit. The detection unit 22 has a screw portion that is screwed with the feed screw 301, and moves in the X-axis direction when the feed screw 301 rotates. The feed screw 301 is disposed in parallel with the straight line F, and when the detection unit 22 is moved by the feed screw 301, the optical axis of the lens 23 of the detection unit 22 coincides with the center of each of the detection units 111a and 111b. Are arranged as follows. After moving to a predetermined position, the detection unit 22 sequentially irradiates the detection units 111a and 111b with excitation light from the lens 23, receives fluorescence emitted from the fluorescent material, and outputs an electrical signal.

  The feed screw 301 is driven via the joint 302 by the detection unit drive motor 61. The detection unit drive motor 61 is a pulse motor, for example, and rotates by a predetermined amount by a pulse. The position sensor 41 is a sensor such as a photo reflector or a mechanical switch provided for detecting the initial position of the detection unit 22.

  The detection unit 22 is provided with a guide hole (not shown in FIG. 4) for preventing rotation, and moves along a guide rod that passes through the guide hole. The guide bar is disposed in parallel with the feed screw 301.

  In this embodiment, the case where two detection units 111a and 111b are provided on the microchip 1 has been described. However, the number of detection units 111 may be any number as long as it is one or more.

  FIG. 5 illustrates a case where the detection unit 22 is in a position where the reaction result of the reagent that occurs in the detection unit 111 (any one of the detection units 111a and 111b) can be optically detected.

  The inlet 110 of the microchip 1 is provided at a position where it communicates with a corresponding opening provided in the packing 90b when the microchip 1 is brought into close contact with the micropump 75 via the packing 90b.

  A driving liquid tank 91 is connected to the suction port 145 of the micro pump unit 75 via a packing 90a, and the driving liquid filled in the driving liquid tank 91 is sucked via the packing 90a. On the other hand, the discharge port 146 communicates with the injection port 110 of the microchip 1 through the packing 90b.

  By driving the piezoelectric element 112, the driving liquid sent out from the micropump unit 75 is injected from the injection port 110 of the microchip 1 into the channel 250 formed in the microchip 1. In this way, the driving liquid is injected from the micropump unit 75 into the injection port 110.

  The micropump unit 75 is provided with at least one micropump. When the microchip 1 illustrated in FIG. 2 is driven, two micropumps corresponding to the two inlets 110a and 110b are necessary.

  FIG. 7 is a circuit block diagram of the detection device 80 in the embodiment of the present invention.

  The control unit 99 includes a CPU 98 (central processing unit), a RAM 97 (Random Access Memory), a ROM 96 (Read Only Memory), and the like, and reads a program stored in the ROM 96 as a nonvolatile storage unit to the RAM 97. Each part of the detection device 80 is centrally controlled according to the program. The control unit 99 is a control means of the present invention.

  Hereinafter, functional blocks having the same functions as those described so far are denoted by the same reference numerals, and description thereof is omitted.

  The CPU 98 includes a pump drive control unit 411, an arrival determination unit 412, a detection unit drive control unit 413, and a threshold value calculation unit 414. The arrival determination unit 412 is an arrival determination unit of the present invention, and the threshold calculation unit 414 is a threshold calculation unit of the present invention.

  Detection signals output from the liquid detection unit 19a and the liquid detection unit 19b, and from the liquid detection unit 20a and the liquid detection unit 20b are converted into digital values by an A / D converter (not shown) built in the control unit 99, and are sent to the CPU 98. Entered.

  The threshold calculation unit 414 calculates the threshold based on a value obtained by delaying the detection signals output from the liquid detection unit 19 and the liquid detection unit 20 by a predetermined time.

  The arrival determination unit 412 determines the presence or absence of liquid based on the detection signal output by the liquid detection unit 20 and the threshold calculated by the threshold calculation unit 414.

  The threshold calculation procedure will be described in detail later.

  The detection unit drive control unit 413 instructs the detection unit drive detection unit drive motor 61 to move the detection unit 22.

  The pump driving unit 500 drives the piezoelectric element 112 of each micropump. Based on the program, the pump drive control unit 411 controls the pump drive unit 500 to inject or suck a predetermined amount of drive fluid. The pump drive unit 500 receives a command from the pump drive control unit 411 and generates a drive voltage to drive the piezoelectric element 112.

  The CPU 98 performs inspections in a predetermined sequence and stores the inspection results in the RAM 97. The inspection result can be stored in the memory card 501 by the operation of the operation unit 87 or printed by the printer 503.

  Next, the procedure in which the inspection apparatus 80 according to the first embodiment of the present invention performs an inspection will be described with reference to FIGS.

  FIG. 8 is a graph illustrating the relationship between the detection signal level X output from the liquid detection unit according to the first embodiment and the threshold value Z calculated by the threshold value calculation unit 414. FIG. 9 is a flowchart for explaining a subroutine by which the inspection apparatus 80 according to the first embodiment calculates a threshold value. FIG. 10 is a flowchart for explaining a subroutine in which the inspection apparatus 80 according to the embodiment of the present invention detects that the liquid leading portion has reached the liquid reservoir. FIG. 11 is a flowchart illustrating a main routine in which the inspection apparatus 80 according to the embodiment of the present invention performs an inspection. FIG. 12 is a flowchart illustrating a routine in which the inspection apparatus 80 according to the embodiment of the present invention performs a reaction measurement.

  First, the threshold calculation procedure will be described with reference to the flowchart of FIG.

  Note that when the threshold value calculation routine is called, one of the liquid detection units 20a and 20b or the liquid detection units 19a and 19b is designated by an argument to calculate the threshold value. In the following description, a case where the liquid detection unit 20a is designated will be described as an example.

  S10: A step of measuring the reflected light from the liquid reservoir.

  The arrival determination unit 412 causes the designated liquid detection unit 20a to emit light, and measures the detection signal level X output from the liquid detection unit 20a that has received the light reflected from the liquid storage unit 140a.

  S11: A step of storing the detection signal level X.

  The arrival determination unit 412 stores the detection signal level X measured in step S10 in the RAM 97.

  S12: A step of waiting for Δt.

  The CPU 98 sets an internal timer and waits for a predetermined time Δt.

  S13: A step of calculating a threshold value Z.

  The threshold calculation unit 414 reads the detection signal level X stored in the RAM 97. Next, the threshold value calculation unit 414 reads a predetermined value Δd stored in advance in the ROM 96 or the RAM 97 and sets a value obtained by subtracting the predetermined value Δd from the detection signal level X as the threshold value Z.

  The process is thus completed, and the process returns to the original routine.

  In FIG. 8A, the horizontal axis is the time axis t, and the vertical axis is the digital value D, which exemplifies output values from the start of inspection of the detection signal level X and the change in the threshold value Z. Since the threshold value Z is calculated according to the above-described procedure, it is output from t1 after Δt from the start of the inspection, and is a value less than the detection signal level X by Δd.

  FIG. 8A shows an example in which the detection signal level X gradually decreases due to condensation in the flow path. In the present embodiment, since the threshold value Z is calculated according to the above-described procedure, the threshold value Z is also reduced in the same manner as the detection signal level X as shown in FIG.

  FIG. 8B shows the determination result of the arrival determination unit 412 as H when the detection signal level X ≧ threshold Z, and L when the detection signal level X <threshold Z. As shown in FIG. 6B, when the timing at which the liquid reaches the liquid reservoir is t2, the detection signal level X rapidly decreases at t2 and is below the threshold value Z. The arrival determination unit 412 detects that the detection signal level X <the threshold Z, and the determination result transitions from H to L as shown in FIG.

  As described above, in the first embodiment, the threshold value Z is delayed by the predetermined time Δt and is reduced in the same manner as the detection signal level X. Therefore, when the detection signal level X is gradually reduced, the detection signal level X <the threshold value Z is not satisfied. . On the other hand, when the detection signal level X decreases rapidly, the detection signal level X <the threshold value Z. Therefore, even when the detection signal level X gradually decreases due to condensation in the flow path, the timing at which the liquid reaches the liquid reservoir can be accurately determined.

  For example, Δt may be about 1 second, and Δd may be about 10 as a digital value when quantized with 8 bits. Further, once the determination result of the arrival determination unit 412 becomes L, it does not become H even if the detection signal level X ≧ threshold Z, unless reset.

  Next, the liquid arrival detection routine will be described with reference to the flowchart of FIG.

  In the following description, the liquid reservoir 140a or the liquid reservoir 141a is used as the a-channel liquid reservoir, the liquid reservoir 140b or the liquid reservoir 141b is used as the b-channel liquid reservoir, and the pump for injecting the driving liquid from the inlet 110a is used. An a-channel pump and a pump that injects the driving liquid from the inlet 110b are referred to as a b-channel pump.

  Note that when the liquid arrival detection routine is called, either the liquid detection unit 20 or the liquid detection unit 19 is designated by an argument to detect the arrival of the liquid. In the following description, a case where the liquid detection unit 20 is designated will be described as an example.

  S101: This is a step of calculating a threshold value Za for determining the arrival of the liquid.

  The CPU 98 calls a threshold value calculation routine to calculate the threshold value Za of the liquid detection unit 20a.

  S102: A step of measuring the reflected light from the liquid reservoir of the a channel.

  The arrival determination unit 412 causes the liquid detection unit 20a to emit light, and measures the detection signal level Ya output from the liquid detection unit 20a that has received the light reflected from the liquid reservoir 140a of the a channel.

  S103: This is a step of comparing the detection signal level Ya with the threshold value Za.

  The arrival determination unit 412 compares the detection signal level Ya proportional to the light reception level of the liquid detection unit 20a with the threshold value Za obtained by the threshold value calculation routine in step S101, and determines whether or not the detection signal level Ya <the threshold value Za. judge.

  When the detection signal level Ya <the threshold value Za (step S102; Yes), the process proceeds to step S103.

  S104: a step of stopping the pump of the a channel.

  The pump drive control unit 411 stops the a-channel pump. When the leading portion of the liquid reaches the liquid reservoir portion of the a channel, the liquid reservoir portion of the a channel is filled with the liquid, and the detection signal level Ya <threshold value Za is satisfied. When this state is detected, the pump drive control unit 411 stops the pump of the a channel and aligns the positions of the liquid channel flowing through the channel with the b channel.

  If the detection signal level Ya ≧ the threshold value Za (step S103; No), the process proceeds to step S105.

  S105: A step of calculating a threshold value Zb for determining the arrival of the liquid.

  The CPU 98 calls a threshold value calculation routine to calculate the threshold value Zb.

  S106: This is a step of measuring the reflected light from the liquid reservoir.

  The arrival determination unit 412 causes the liquid detection unit 20b to emit light, and measures the detection signal level Yb output from the liquid detection unit 20b that has received the light reflected from the b-channel liquid reservoir.

  S107: This is a step of comparing the detection signal level Yb with the threshold value Zb.

  The arrival determination unit 412 compares the detection signal level Yb proportional to the light reception level of the liquid detection unit 20b with the threshold value Zb obtained when the calibration is performed in step S201, and whether or not the detection signal level Yb <the threshold value Zb. Whether or not.

  If the detection signal level Yb <the threshold value Zb (step S105; Yes), the process proceeds to step S108.

  S108: A step of stopping the pump of the b channel.

  The pump drive control unit 411 stops the b-channel pump. When the leading portion of the liquid reaches the liquid storage part of the b channel, the liquid storage part of the b channel is filled with the liquid, and the detection signal level Yb <the threshold value Zb. When this state is detected, the pump drive control unit 411 stops the b-channel pump, and aligns the leading positions of the liquid flowing through the a-channel and the flow path.

  If the detection signal level Yb ≧ the threshold value Zb (step S105; No), the process proceeds to step S109.

  S109: This is a step of determining whether or not the pumps of both channels a and b are stopped.

  The pump drive control unit 411 determines whether or not the pumps of both channels a and b are stopped.

  When the pumps of both channels a and b are stopped (step S109; Yes), the process is terminated and the process returns to the original routine.

  When the a or b channel pump is not stopped (step S109; No), the process proceeds to step S110.

  S110: It is a step which determines whether it is time-out.

  The CPU 98 reads the elapsed time from the internal timer and determines whether or not the timeout time has elapsed.

  In the case of timeout (step S110; Yes), the process proceeds to step S111.

  S111: This is a step of setting the error flag to 1.

  The CPU 98 sets the error flag to 1 and returns to the original routine.

  When it is not time-out (step S110; No), it returns to step S101.

  If not timed out, the process is continued and the liquid is waited for to reach the liquid reservoir.

  This is the end of the description of the liquid arrival routine.

  Next, an outline procedure of inspection will be described in the order of the flowchart of FIG.

  Assume that the microchip 1 is set at a position where inspection can be performed as shown in FIG. 5, and the CPU is instructed to start inspection by operating the operation unit 87. The detection unit 22 is assumed to be in the initial position shown in FIG.

  S201: This is a step for performing calibration.

  The arrival determination unit 412 causes the liquid detection unit 20a and the liquid detection unit 20b to emit light, and outputs the output signals detected by the liquid detection unit 20a and the liquid detection unit 20b respectively from the light reflected from the liquid storage unit 140a and the liquid storage unit 140b. Compare with a predetermined signal level. If it is not within the predetermined signal level range, the arrival determination unit 412 performs calibration so that the current flowing through the light emitting units of the liquid detection unit 20a and the liquid detection unit 20b is increased or decreased to be within the predetermined signal level range. Also, the error flag is initialized and the value is set to zero.

  S202: This is a step of starting the pump.

  The pump drive control unit 411 instructs the pump drive unit 500 to send liquid from the micropump unit 75 to the microchip 1.

  S203: This is a step of detecting the arrival of the liquid.

  The arrival determination unit 412 calls a liquid arrival detection routine which will be described in detail later, detects that the liquid leading portion has reached the liquid reservoir 140a and the liquid reservoir 140b, and stops driving the pump. In this step, since the leading position of the reagent or the like flowing through the flow channels of the a channel and the b channel can be aligned with the liquid reservoir 140a and the liquid reservoir 140b, the mixing unit 130a and the mixing unit are started when the pump is started in step 208 later. A reagent or the like can be simultaneously injected into 130b.

  S204: This is a step of determining an error flag.

  The arrival determination unit 412 determines an error flag. When the liquid arrival detection routine is normally completed within a predetermined time, the error flag is 0, and the micropump unit 75 is stopped while the liquid has reached the liquid reservoir 140a and the liquid reservoir 140b. On the other hand, the error flag is set to 1 when the process does not end normally within a predetermined time.

  When the error flag is 1 (step S204; Yes), the process proceeds to step S205.

  S205: This is a step of displaying a warning.

  The control unit 99 displays an error warning on the display unit 84 and stops the inspection apparatus 80.

  When the error flag is 0 (step S204; No), the process proceeds to step S206.

  S207: This is a step for performing calibration.

  The arrival determination unit 412 causes the liquid detection unit 20a and the liquid detection unit 20b to emit light, and the output signals detected by the liquid detection unit 20a and the liquid detection unit 20b respectively from the light reflected from the liquid storage unit 141a and the liquid storage unit 141b. Compare with a predetermined signal level. If it is not within the predetermined signal level range, the arrival determination unit 412 performs calibration so that the current flowing through the light emitting units of the liquid detection unit 20a and the liquid detection unit 20b is increased or decreased to be within the predetermined signal level range. Also, the error flag is initialized and the value is set to zero.

  S208: This is a step of starting the pump.

  The pump drive control unit 411 instructs the pump drive unit 500 to send liquid from the micropump unit 75 to the microchip 1.

  S209: This is a step of detecting the arrival of the liquid.

  The arrival determination unit 412 calls the liquid arrival detection routine, detects that the liquid has reached the liquid reservoir 141a and the liquid reservoir 141b, and stops driving the pump. In this step, since the head portion of the mixed liquid of the specimen and the reagent flowing through the channel of the a channel and the b channel is aligned with the liquid reservoir 141a and the liquid reservoir 141b, the mixed liquid is filled in the detector 111a and the detector 111b. Can be in a state.

  S210: This is a step of determining an error flag.

  The arrival determination unit 412 determines an error flag. When the liquid arrival detection routine ends normally within a predetermined time, the error flag is 0, and the micropump unit 75 is stopped in a state where the liquid has reached the liquid reservoir 141a and the liquid reservoir 141b. On the other hand, the error flag is set to 1 when the process does not end normally within a predetermined time.

  When the error flag is 1 (step S210; Yes), the process proceeds to step S211.

  S211: A step of displaying a warning.

  The control unit 99 displays an error warning on the display unit 84 and stops the inspection apparatus 80.

  When the error flag is 0 (step S210; No), the process proceeds to step S212.

  S213: This is a step of measuring the reaction result.

  After a predetermined time has elapsed, the CPU 98 calls a reaction measurement routine and measures the reaction results from the detection unit 111a and the detection unit 111b.

  This is the end of the description of the main routine.

  Next, the reaction measurement routine will be described in the order of the flowchart of FIG.

  S301: This is a step of moving the detection unit 22.

  The detection unit drive control unit 413 sends a predetermined number of pulses to the detection unit drive motor 61 to move the detection unit 22 in the direction of the arrow S2 in FIG. 4, and the detection unit 22 detects the reaction result of the detection unit 111a. Stop.

  S302: It is a step which measures a reaction result.

  After a predetermined time has elapsed, the CPU 98 causes the light emitting section of the detection unit 22 to emit light, measures the output signal level from the detection unit 22, and stores the result in the RAM 97.

  S303: This is a step of moving the detection unit 22.

  The detection unit drive control unit 413 sends a predetermined number of pulses to the detection unit drive detection unit drive motor 61 to further move the detection unit 22 in the direction of the arrow S2 in FIG. 4, and to a position where the detection result of the detection unit 111b is detected. Stop.

  S304: It is a step which measures a reaction result.

  The CPU 98 causes the light emitting unit of the detection unit 22 to emit light, measures the output signal level from the detection unit 22, and stores the result in the RAM 97.

  The inspection is finished as described above, and the detection unit drive control unit 413 returns the detection unit 22 to the initial position.

  This completes the description of the procedure of the reaction measurement routine.

  Next, a second embodiment will be described. In the inspection apparatus 80 according to the first embodiment described so far, the liquid delivery is controlled by detecting that the liquid has reached the liquid reservoir. In the second embodiment, an example will be described in which it is detected that the liquid filled in the liquid reservoir has moved away.

  FIG. 13 is a graph for explaining the relationship between the detection signal level X output from the liquid detection unit according to the second embodiment and the threshold value Z calculated by the threshold value calculation unit 414. FIG. 14 is a flowchart for explaining a subroutine by which the inspection apparatus 80 according to the second embodiment calculates a threshold value.

  In FIG. 13A, the horizontal axis is the time axis t, and the vertical axis is the digital value D, which exemplifies output values from the start of inspection of the detection signal level X and the change in the threshold value Z.

  A difference from FIG. 9A is that the detection signal level X gradually increases in FIG. 13A due to the disappearance of condensation in the flow path. Further, when the liquid filled in the liquid reservoir is moved away, the detection signal level X increases rapidly at the timing t2 as shown in FIG. As in the first embodiment, the threshold value Z output from t1 after a predetermined time Δt from the start of the inspection is a value Δd greater than the detection signal level X in this embodiment.

  FIG. 13B shows the determination result of the arrival determination unit 412 as H when the detection signal level X ≧ threshold Z, and L when the detection signal level X <threshold Z. As described above, when the liquid filled in the liquid reservoir moves and disappears, the detection signal level X increases rapidly.

  In FIG. 13A, the detection signal level X increases rapidly at the timing of t2 and exceeds the threshold value Z. When the detection signal level X ≧ threshold Z at the timing of t2, the determination of the arrival determination unit 412 transitions from L to H as shown in FIG.

  As described above, in the second embodiment, the threshold value Z increases in the same manner as the detection signal level X with a delay of the predetermined time Δt. Therefore, when the detection signal level X gradually increases, the detection signal level X ≧ the threshold value Z does not hold. On the other hand, when the detection signal level X increases rapidly, the detection signal level X ≧ the threshold value Z. Therefore, even when the detection signal level X gradually increases due to the disappearance of condensation in the flow path, the timing at which the liquid reaches the liquid reservoir can be accurately determined.

  For example, Δt may be about 1 second, and Δd may be about 10 as a digital value when quantized with 8 bits. Further, once the determination result of the arrival determination unit 412 becomes L, it does not become H even if the detection signal level X ≧ threshold Z, unless reset.

  The threshold value calculation procedure of the second embodiment will be described with reference to the flowchart of FIG.

  Note that when the threshold value calculation routine is called, one of the liquid detection units 20a and 20b or the liquid detection units 19a and 19b is designated by an argument to calculate the threshold value. In the following description, a case where the liquid detection unit 20a is designated will be described as an example. The only difference from the threshold calculation procedure of the first embodiment described in FIG. 9 is the step of calculating the threshold Z of S23, and the same steps as those of the first embodiment are given the same numbers.

  S10: A step of measuring the reflected light from the liquid reservoir.

  The arrival determination unit 412 causes the designated liquid detection unit 20a to emit light, and measures the detection signal level X output from the liquid detection unit 20a that has received the light reflected from the liquid reservoir.

  S11: A step of storing the detection signal level X.

  The arrival determination unit 412 stores the detection signal level X measured in step S10 in the RAM 97.

  S12: A step of waiting for Δt.

  The CPU 98 sets an internal timer and waits for a predetermined time Δt.

  S23: A step of calculating a threshold value Z.

  The threshold calculation unit 414 reads the detection signal level X stored in the RAM 97. Next, the threshold value calculation unit 414 reads a predetermined value Δd stored in advance in the ROM 96 or RAM 97 and sets a value obtained by adding the predetermined value Δd to the detection signal level X as the threshold value Z.

  The process is thus completed, and the process returns to the original routine.

  Next, a third embodiment will be described. The difference between the third embodiment, the first embodiment, and the second embodiment is only the step of calculating the threshold value Z in S23 of the threshold value calculation procedure.

  The threshold value calculation procedure of the third embodiment will be described with reference to the flowchart of FIG.

  Note that when the threshold value calculation routine is called, one of the liquid detection units 20a and 20b or the liquid detection units 19a and 19b is designated by an argument to calculate the threshold value. In the following description, a case where the liquid detection unit 20a is designated will be described as an example. In addition, the same number is attached | subjected to the process same as 1st Embodiment.

  S10: A step of measuring the reflected light from the liquid reservoir.

  The arrival determination unit 412 causes the designated liquid detection unit 20a to emit light, and measures the detection signal level X output from the liquid detection unit 20a that has received the light reflected from the liquid reservoir.

  S11: A step of storing the detection signal level X.

  The arrival determination unit 412 stores the detection signal level X measured in step S10 in the RAM 97.

  S12: A step of waiting for Δt.

  The CPU 98 sets an internal timer and waits for a predetermined time Δt.

  S33: A step of calculating a threshold value Z.

  The threshold calculation unit 414 reads the detection signal level X stored in the RAM 97. Next, the threshold value calculation unit 414 reads a predetermined coefficient α stored in the ROM 96 or the RAM 97 and sets a value obtained by multiplying the detection signal level X by the coefficient α as the threshold value Z.

  The process is thus completed, and the process returns to the original routine.

  As described above, in the third embodiment, the threshold value calculation unit 414 calculates the threshold value Z by multiplying the detection signal level X by the predetermined coefficient α. When detecting that the liquid has reached and filled the liquid reservoir as in the first embodiment, the coefficient α is set to an appropriate value in the range of 0 <α <1 and stored in the ROM 96 or RAM 97 in advance. The threshold calculation unit 414 reads in step S33. Further, when it is detected that the liquid filled in the liquid reservoir has moved and disappeared as in the second embodiment, the coefficient α is similarly set to an appropriate value exceeding 1 in advance, and the ROM 96 or RAM 97 is similarly set. Remember me.

  In the third embodiment, since the threshold value Z is changed at a constant rate with respect to the detection signal level X, erroneous determination is made even when the detection signal level X varies greatly until the liquid reaches or disappears from the liquid reservoir. Can be prevented.

  As described above, according to the present invention, it is possible to provide an inspection apparatus capable of correctly detecting the liquid in the fine flow path even when the temperature of the microchip changes.

It is an external view of the test | inspection apparatus 80 in embodiment of this invention. It is explanatory drawing of the microchip 1 concerning embodiment of this invention. It is an enlarged view of the periphery of the detection part 111. It is a perspective view which shows an example of the internal structure of the test | inspection apparatus 80 in embodiment of this invention. It is sectional drawing which shows an example of the internal structure of the test | inspection apparatus 80 in embodiment of this invention. It is explanatory drawing explaining the state with which the detection part 111 is filled with the liquid. It is a circuit block diagram of the detection apparatus 80 in embodiment of this invention. 6 is a graph for explaining a relationship between a detection signal level X output from the liquid detection unit according to the first embodiment and a threshold value Z calculated by a threshold value calculation unit 414; It is a flowchart explaining the subroutine with which the test | inspection apparatus 80 of 1st Embodiment calculates a threshold value. It is a flowchart explaining the subroutine with which the test | inspection apparatus 80 of embodiment of this invention detects that the head part of the liquid reached | attained the liquid reservoir. It is a flowchart explaining the main routine which the test | inspection apparatus 80 of embodiment of this invention performs a test | inspection. It is a flowchart explaining the routine with which the test | inspection apparatus 80 of embodiment of this invention performs reaction measurement. It is a graph explaining the relationship between the detection signal level X which the liquid detection part which concerns on 2nd Embodiment outputs, and the threshold value Z which the threshold value calculation part 414 calculated. It is a flowchart explaining the subroutine with which the test | inspection apparatus 80 of 2nd Embodiment calculates a threshold value. It is a flowchart explaining the subroutine with which the test | inspection apparatus 80 of 3rd Embodiment calculates a threshold value.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Microchip 19, 20 Liquid detection part 21 Detection board 22 Detection unit 61 Detection unit drive motor 75 Micropump unit 80 Inspection apparatus 82 Case 83 Insertion port 84 Display part 87 Operation button 90 Packing 110 Inlet 111 Detection part 121 Sample accommodation Units 120, 122, 123 Reagent storage unit 130, 131 Mixing unit 140, 141 Liquid reservoir unit 152, 153 Temperature control unit 250 Channel 411 Pump drive control unit 412 Arrival determination unit 413 Detection unit drive control unit 414 Threshold calculation unit

Claims (3)

In an inspection device for measuring the result of moving the sample and reagent stored in the flow path of the microchip and filling the detection unit and reacting them,
Liquid detection means for outputting a detection signal corresponding to the amount of liquid that has reached a predetermined position of the flow path;
Threshold calculation means for calculating a threshold based on a value obtained by delaying the detection signal for a predetermined time;
Arrival determination means for determining the presence or absence of the liquid based on the detection signal and the threshold;
An inspection apparatus comprising: The threshold calculation means includes
The inspection apparatus according to claim 1, wherein a threshold value is calculated by adding or subtracting a predetermined value to a value obtained by delaying the detection signal for a predetermined time. The threshold calculation means includes
The inspection apparatus according to claim 1, wherein a threshold value is calculated by multiplying a value obtained by delaying the detection signal by a predetermined time by a predetermined value.

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