JP2009097953A - Examination apparatus - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an examination apparatus capable of detecting, without contact, the presence or the absence of a small amount of specimen injected to a microchip, using a simple procedure. <P>SOLUTION: In the examination apparatus allows the specimen to be injected to the microchip for reacting to a reagent and measures the reaction result, a specimen detection means for detecting the specimen injected to the microchip without contacts is provided. <P>COPYRIGHT: (C)2009,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 called μ-TAS (Micro total Analysis System), bioreactor, lab-on-chip, biochip, medical test / diagnosis field, environmental measurement field, Its application is expected in the field of 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 the driving liquid into the microchannel by a micropump, moves the reagent, etc., and flows it to the reaction unit and then to the detection unit, whereby blood Have proposed a test apparatus capable of measuring a reaction result with a specimen (see, for example, Patent Document 2).

  In such an inspection apparatus, the inspection apparatus automatically reacts a microchip into which a specimen has been previously injected with a reagent or the like, and measures the reaction result. However, if the tester forgets to inject the sample into the microchip, or if the sample is not injected due to an erroneous operation, the test device measures the reaction result in a predetermined procedure, so the test result is misjudged. was there.

  On the other hand, by using a reagent containing a colored substance or a fluorescent substance, a method of simply confirming the presence of the reagent when dispensing the reagent into each reaction part has been proposed (for example, see Patent Document 3).

Also, before starting the measurement, the remaining amount of reagent in the reagent bottle corresponding to the measurement item is detected by the reagent remaining amount detection means, and the inspection device accurately recognizes it, thereby avoiding problems due to reagent shortage during analysis Is disclosed (see, for example, Patent Document 4).
JP 2004-28589 A JP 2006-149379 A JP 2006-346613 A JP-A-6-66813

  However, as disclosed in Patent Document 3, when a sample is colored or fluorescent and visually confirmed, a step of mixing a colored substance or a fluorescent material with the sample in advance and a visual confirmation step are required. , The procedure for inspection will increase.

  Further, in the method disclosed in Patent Document 4, since the remaining amount of the reagent in the reagent bottle is detected by a sensor provided in the nozzle, the presence / absence of a minute amount of sample injected into the microchip is detected. Not applicable to

  The present invention has been made in view of the above problems, and an object of the present invention is to provide a test apparatus capable of detecting the presence or absence of a minute amount of sample injected into a microchip by a simple procedure in a non-contact manner. To do.

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

1.
In an inspection device that injects a sample into a microchip, reacts the sample with a reagent, and measures the reaction result.
An inspection apparatus comprising: a sample detection means for detecting the sample injected into the microchip in a non-contact manner.

2.
Sample determination means for determining the presence or absence of the sample based on the detection output of the sample detection means;
Control means for controlling the inspection device;
Have
2. The inspection apparatus according to 1, wherein the control unit performs control based on determination by the specimen determination unit.

3.
Display means for displaying warning information;
3. The inspection apparatus according to 2, wherein the control unit displays the warning information on the display unit based on the determination of the sample determination unit.

  According to the present invention, it is possible to detect the presence or absence of a minute amount of sample injected into a microchip by a simple procedure without contact.

  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, 88 denotes an input / output terminal, and 89 denotes a power switch.

  After the inspection person turns on the power switch 89, the microchip 1 is inserted in the direction of the arrow in FIG. 1, and the operation panel 87 is operated 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.

  2A and 2B are external views of the microchip 1. FIG. In FIG. 2A, an arrow indicates an insertion direction in which the microchip 1 is inserted into the inspection apparatus 80, and FIG. 2A illustrates a surface that becomes the lower surface of the microchip 1 when inserted. FIG. 2B is a side view of the microchip 1.

  The window 111a and the channel 111b of the detection unit 111 in FIG. 2A are provided for optically detecting the reaction between the specimen and the reagent, and are made of a transparent member such as glass or resin. Reference numerals 110a, 110b, 110c, 110d, and 110e denote driving liquid injection units that communicate with the internal fine flow paths. The driving liquid injection units 110 inject driving liquids to drive internal reagents and the like.

  Reference numeral 113 denotes a sample injection unit for injecting a sample into the microchip 1. A sample such as blood is injected from the sample injection unit 113 by using a syringe or the like, and the sample is stored in the connected sample storage unit 121.

When optically detecting a sample stored in the sample storage unit 121, at least the window 121a of the sample storage unit 121 is formed of a transparent member such as glass or resin as shown in FIG. When detecting from the light transmitted through the specimen storage unit 121, the flow path 121b is also formed of a transparent member such as glass or resin.
Note that when a specimen is detected using ultrasonic waves, radio waves, or the like, it is not always necessary to use a transparent member.

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

  The 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.

  In particular, it is desirable that a chip intended for a large number of measurement specimens, particularly clinical specimens at risk of contamination and infection, be of a disposable type. Therefore, a plastic resin that can be mass-produced, is lightweight, is strong against impact, and can be easily disposed of by incineration, for example, polystyrene that is excellent in transparency, mechanical properties, and moldability and is easy to be finely processed is preferable. For example, when it is necessary to heat the chip to near 100 ° C. in analysis, it is preferable to use a resin having excellent heat resistance (for example, polycarbonate). In addition, when protein adsorption becomes a problem, it is preferable to use polypropylene. Resin and glass have low thermal conductivity, and by using these materials in the locally heated region of the microchip, heat conduction in the surface direction is suppressed, and only the heated region is selectively heated. Can do.

  Since the detection unit 111 optically detects a color reaction product, a fluorescent substance, and the like, at least the substrate of this part uses a light-transmitting material (for example, alkali glass, quartz glass, transparent plastics), It is necessary to transmit light. The window 111a of the detection unit 111 and at least the groove forming substrate that forms the flow path of the detection unit 111 are made of a light-transmitting material, and transmit light through the detection unit 111.

  When optically detecting the sample stored in the sample storage unit 121 so as to transmit light through the sample storage unit 121, the same material as that of the detection unit 111 is used. A groove forming substrate for forming the window 121a and the channel 121b is formed.

  The microchip 1 is provided with minute groove-like channels (microchannels) and functional parts (channel elements) for performing inspection, sample processing, and the like in an appropriate manner according to the application. ing. In the present embodiment, an example of a process for performing amplification and detection of a specific gene performed in the microchip 1 by using these microchannels and channel elements will be described with reference to FIG.

  FIG. 2C is an explanatory diagram for explaining an example of the functions of the micro flow channel and the flow channel element inside the microchip 1.

  For example, a sample storage unit 121 that stores a sample liquid, and reagent storage units 120a, 120b, 120c, and 120d that store reagents are provided in the microchannel, and can be quickly examined regardless of location or time. As described above, necessary reagents, a cleaning solution, a denaturing solution, and the like are stored in advance in the reagent storage units 120a, 120b, 120c, and 120d. In FIG. 2C, the reagent storage units 120a, 120b, 120c, and 120d, the sample storage unit 121, and the flow path element are represented by squares, and the fine flow path therebetween is represented by a solid line and an arrow.

  The microchip 1 includes a groove forming substrate 108 in which a fine flow path is formed and a covering substrate 109 that covers the groove-shaped flow path. The fine channel is formed on the order of micrometers, 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.

  At least in the groove forming substrate 108 of the microchip 1, the fine flow path is formed. The coated substrate 109 needs to cover at least the fine flow path of the groove forming substrate in close contact, and may cover the entire surface of the groove forming substrate. Note that the microchannel 1 is provided with a part for controlling liquid feeding, such as a liquid feeding control unit (not shown), a backflow prevention unit (a check valve, an active valve, etc.), and the like. In this case, liquid feeding is performed according to a predetermined procedure.

  The sample injection unit 113 is an injection unit for injecting the sample into the microchip 1, and the driving liquid injection unit 110 is an injection unit for injecting the driving liquid 11 into the microchip 1. Prior to performing the test using the microchip 1, the tester injects the sample from the sample injection unit 113 using a syringe or the like. As shown in FIG. 2C, the sample injected from the sample injection unit 113 is stored in the sample storage unit 121 through the communicating fine channel.

  Next, when the driving liquid is injected from the driving liquid injection unit 110 a, the driving liquid 11 pushes the sample stored in the sample storage unit 121 through the communicating fine flow path, and sends the sample to the amplification unit 122.

  On the other hand, the driving liquid injected from the driving liquid injection unit 110b pushes out the reagent stored in the reagent storage unit 120a through the communicating fine channel. The reagent pushed out from the reagent storage unit 120a is sent to the amplification unit 122 by the driving liquid. Depending on the reaction conditions at this time, it is necessary to set the amplification unit 122 to a predetermined temperature, and as described later, the reaction is performed at a predetermined temperature by heating or absorbing heat inside the inspection device 80.

  After a predetermined reaction time, a solution containing the sample after reaction sent out from the amplification unit 122 by the driving liquid 11 is injected into the detection unit 111. Subsequently, when the driving liquid 11 is injected from the driving liquid injection sections 110c, 110d, and 110e, the driving liquid 11 pushes out the reagent stored in the reagent storage sections 120b, 120c, and 120d through the communicating fine flow path, and detects the detection section. 111 is injected.

  When the detection unit 111 is irradiated with light from the window 111, the reagent that has reacted with the specimen emits, for example, fluorescence. Therefore, the reaction result can be measured by measuring the amount of fluorescence. The solution after the reaction result is measured by the detection unit 111 is sent to the waste liquid storage unit 125.

  FIG. 3 is a cross-sectional view showing an example of the internal configuration of the inspection apparatus 80 according to the first embodiment. The inspection apparatus 80 includes a temperature adjustment unit 152, a light detection unit 150, a specimen detection unit 155, an intermediate flow path unit 180, a micro pump unit 75, packings 90a and 90b, a chip connection unit 8, a driving liquid tank 91, and the like. Hereinafter, the same reference numerals are given to the same components as those described so far, and description thereof will be omitted.

  FIG. 3 shows a state in which the lower surface of the microchip 1 is brought into close contact with the chip connection portion 8 so that the driving liquid injection port 110 and the flow path opening 185 of the chip connection portion 8 communicate with each other. The microchip 1 is placed on a chip transport tray (not shown in FIG. 3), and moves in the left-right direction on the paper surface as the chip transport tray moves. When the microchip 1 moves to a position where the driving liquid injection port 110 and the flow path opening 185 can communicate with each other, the temperature adjustment unit 152 moves downward in the drawing and presses the upper surface of the microchip 1 to connect the driving liquid injection port 110 with the driving liquid injection port 110. The channel opening 185 is brought into close contact.

  The temperature adjustment unit 152 is a unit that incorporates a Peltier element, a power supply device, a temperature control device, and the like and adjusts the upper surface of the microchip 1 to a predetermined temperature by generating heat or absorbing heat.

  When the microchip 1 is transported to a predetermined position, the detection unit 95 detects the chip transport tray 2 and is turned on.

  The sample detection unit 155 includes a second light emitting unit 155a using an LED or a lamp, and a second light receiving unit 155b using a photoelectric conversion element such as a photodiode. The sample detection unit 155 transmits light transmitted through the sample storage unit 121 of the microchip 1. It is arranged so that it can be detected. In the present embodiment, an example in which light transmitted through the sample storage unit 121 is detected will be described. However, ultrasonic waves or radio waves transmitted through the sample storage unit 121 may be detected.

  The driving liquid injection port 110 of the microchip 1 is provided at a position that communicates with a corresponding flow path opening 185 provided in the chip connection portion 8 when the microchip 1 and the chip connection portion 8 are brought into close contact with each other. . The intermediate flow path portion 180 includes a transparent first substrate 184 provided with a groove of the intermediate flow path 182 and a transparent second substrate 183 that covers the first substrate 184. One end of the intermediate flow path 182 has a flow path opening. The other end communicates with the flow path opening 186 of the packing 90b. The intermediate flow path 182 communicates with the input / output port 146 of the micro pump unit 75 through the flow path opening 186.

  A driving liquid tank 91 is connected to the suction side of the micropump 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 input / output port 146 provided on the discharge-side end face of the micropump unit 75 communicates with the driving liquid injection port 110 of the microchip 1 via the intermediate flow path 182, and thus is sent out from the micropump unit 75. The driving liquid thus injected is injected from the driving liquid injection port 110 of the microchip 1 into the flow path 250 formed in the microchip 1. In this way, the driving liquid is injected from the micropump unit 75 into the driving liquid injection port 110.

  The micropump unit 75 is provided with at least one micropump. When the microchip 1 illustrated in FIG. 2 is driven, five micropumps corresponding to the five driving liquid inlets 110a, 110b, 110c, 110d, and 110e are necessary. In addition, corresponding five flow path openings 186, intermediate flow paths 182 and flow path openings 185 are required so that the five micropumps communicate with the five driving liquid inlets 110a, 110b, 110c, 110d, and 110e, respectively. .

  In the detection unit 111 of the microchip 1, the specimen and the reagent stored in the microchip 1 react to cause, for example, coloration, light emission, fluorescence, turbidity, and the like. In the present embodiment, as described with reference to FIG. 2, the reaction result of the reagent that occurs in the detection unit 111 is optically detected. The light detection unit 150 includes a first light emitting unit 150a and a first light receiving unit 150b, and is arranged so that light transmitted through the detection unit 111 of the microchip 1 can be detected.

  FIG. 4 is a circuit block diagram of the reaction 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 reaction detector 80 is centrally controlled according to the program.

  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 and a sample determination unit 412.

  The sample determination unit 412 determines the result by comparing the output signal of the second light receiving unit 155b, which detects the light transmitted through the sample storage unit 121 with the light emitted from the second light emitting unit 155a, with a predetermined signal level.

  The pump drive unit 500 is a drive unit that 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.

  FIG. 5 is a flowchart for explaining the procedure of the inspection performed by the inspection apparatus 80 according to the embodiment of the present invention.

  Assume that the microchip 1 is set at a position where inspection can be performed as shown in FIG. 3, and the CPU is instructed to start inspection by operating the operation button 87.

  S101: This is a step of determining whether or not a sample has been injected.

  The specimen determination unit 412 causes the second light emitting unit 155a to emit light and determines whether or not the output signal level of the second light receiving unit 155b is equal to or higher than a predetermined level. As the determination level, for example, a value obtained by actually measuring the output signal level when the amount of the sample is 90% of the prescribed amount is stored in the ROM 96.

  If it is below the predetermined level (step S101; No), the process proceeds to step S110.

  S110: This is a step of displaying a warning.

  The CPU 98 displays a warning that the amount of the sample is insufficient on the display unit 84, and proceeds to step 104 where the inspection apparatus 80 is stopped.

  If the level is equal to or higher than the predetermined level (step S101; Yes), the process proceeds to step S102.

  S102: This is the step of feeding liquid.

  The pump drive control unit 411 instructs the pump drive unit 500 to send liquid from the micropump to the microchip 1 in a predetermined order.

  S103: It is a step which measures a reaction result.

  After a predetermined time has elapsed, the CPU 98 causes the first light emitting unit 150a to emit light, measures the output signal level of the first light receiving unit 150b, and stores the result in the RAM 97.

  S104: This is a step of stopping the inspection apparatus 80.

  The CPU 98 stops each part of the inspection device 80.

  FIG. 6 is a cross-sectional view illustrating an example of the internal configuration of the inspection apparatus 80 according to the second embodiment. The difference between the first embodiment and the second embodiment is that the sample detection unit 156 detects light reflected from the sample storage unit 121. As the specimen detection unit 156, a photoelectric conversion element such as a photodiode or an imaging element such as a CCD can be used. Alternatively, the specimen storage unit 121 may be illuminated from the specimen detection unit 156 side. The procedure for the inspection apparatus 80 to perform the inspection is exactly the same as the procedure described in FIG.

  The other functional elements are exactly the same, and are given the same numbers and are not described here.

  As described above, according to the present invention, it is possible to provide an inspection apparatus capable of detecting the presence or absence of a minute amount of sample injected into a microchip by a simple procedure in a non-contact manner.

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 explanatory drawing which shows an example of an internal structure of the test | inspection apparatus 80 of 1st Embodiment. It is a circuit block diagram of the inspection apparatus 80 in the 1st Embodiment of this invention. It is a flowchart explaining the procedure of the test | inspection of embodiment of this invention. It is explanatory drawing which shows an example of the internal structure of 1st Embodiment.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Microchip 75 Micro pump unit 80 Inspection apparatus 82 Case 83 Insertion port 84 Display part 87 Operation button 90 Packing 110 Drive liquid injection port 111 Detection part 121 Specimen storage part 121a Window 121b Flow path 150 Photodetection part 155, 156 Specimen detection Part 152 Temperature adjustment unit 180 Intermediate flow path

Claims (3)

In an inspection device that injects a sample into a microchip, reacts the sample with a reagent, and measures the reaction result.
An inspection apparatus comprising: a sample detection means for detecting the sample injected into the microchip in a non-contact manner. Sample determination means for determining the presence or absence of the sample based on the detection output of the sample detection means;
Control means for controlling the inspection device;
Have
The inspection apparatus according to claim 1, wherein the control unit performs control based on determination by the specimen determination unit. Display means for displaying warning information;
The inspection apparatus according to claim 2, wherein the control unit displays the warning information on the display unit based on the determination of the sample determination unit.

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