JP2005164425A - Apparatus for inspecting organism-related substance, inspection stage, and inspection container - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an inspection apparatus of an organism-related substance for properly suppressing heat radiation from an observation window. <P>SOLUTION: The inspection apparatus comprises a temperature adjustment mechanism 111 for adjusting the temperature of a fitted inspection container 101, an observation window 112 for achieving optical observation of a solid phase carrier 102 in the inspection container 101, an observation apparatus 150 for acquiring the optical image of the solid phase carrier 102 in the inspection container 101 from the upper part via the observation window 112, and a pump 115 that is connected to a channel 103 in the fitted inspection container 101 via a tube 114 in terms of fluid. The observation window 112 has two transparent plates 113a, 113b that are arranged with an interval. An inspection stage, including the temperature adjustment mechanism 111, comprises a channel 123 connected to space 125 between the two transparent plates 113a, 113b, and a pump 120 for circulating a transparent liquid to the channel 123. The channel 123 has a heat exchange section 124 for heating the transparent liquid circulated in the channel 123. <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

  The present invention relates to a biologically related substance inspection apparatus, an inspection stage, and an inspection container.

For example, a DNA chip analyzer is known as a biologically relevant substance inspection apparatus. In a DNA chip analyzer, an oligo DNA probe spotted on a DNA chip is reacted with a fluorescently labeled sample solution to detect a signal as fluorescence intensity. For example, JP-A-8-166387 discloses a reaction vessel for a DNA chip using glass or silicon as a substrate. Japanese Patent No. 3208390 and Japanese Patent Application Laid-Open No. 09-504864 disclose DNA chips formed by immobilizing a probe on a porous material substrate. This signal detection is performed by irradiating the DNA chip with excitation light and detecting fluorescence using a photomultiplier or a CCD camera.
JP-A-8-166387 Japanese Patent No. 3208390 JP 09-504864 A

  In order to perform an appropriate test using the above-described DNA chip, it is preferable to control the temperature of the sample liquid, and in general, heating is often performed. Therefore, in the inspection apparatus having a window for observing the DNA chip, the apparatus for inspecting the DNA chip disclosed in Japanese Patent No. 3208390 and Japanese Patent Application Laid-Open No. 09-504864 is disclosed. The temperature of the sample liquid will decrease. As a result, when the temperature of the sample solution is lowered, the desired temperature cannot be reached, so that a desired reaction may not be performed, and improvement in test accuracy is required.

  The present invention has been made in consideration of such actual situations, and its main purpose is to provide a testing technique for biologically relevant substances with high testing accuracy.

  Therefore, a secondary object of the present invention is to provide a biologically related substance inspection apparatus in which heat radiation from the observation window is satisfactorily suppressed. Another secondary object of the present invention is to provide a bio-related substance inspection stage in which heat radiation from the observation window is satisfactorily suppressed.

  The present invention is directed, in part, to a biologically related substance testing apparatus. The inspection apparatus of the present invention includes a temperature adjustment mechanism for adjusting the temperature of an inspection container containing a solid phase carrier on which a probe for detecting a biological substance is solid-phased, and the optical property of the solid phase carrier. An observation window that enables observation, an observation device for acquiring an optical image of the solid phase carrier in the cuvette through the observation window, and an observation window heating unit that directly heats the observation window are provided.

  The present invention is directed, in part, to an inspection stage for biological materials. The inspection stage of the present invention includes a temperature control mechanism for adjusting the temperature of an inspection container containing a solid phase carrier on which a probe for detecting a biological substance is solid-phased, and an optical phase of the solid phase carrier. An observation window that enables observation and observation window heating means that directly heats the observation window are provided.

  ADVANTAGE OF THE INVENTION According to this invention, the test | inspection technique of a biological relevant substance with high test | inspection precision is provided. Thereby, better optical observation of the solid phase carrier can be expected.

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

First Embodiment The present embodiment is directed to a biologically related substance inspection apparatus. FIG. 1 is a schematic cross-sectional view of an inspection apparatus and an inspection container attached to the inspection apparatus according to an embodiment of the present invention, showing a state in which the liquid level of a sample solution is positioned on a solid phase carrier. . FIG. 2 is a schematic cross-sectional view of the same inspection apparatus and inspection container as FIG. 1 and shows a state in which the liquid level of the sample solution is located below the solid phase carrier.

  As shown in FIG. 1 and FIG. 2, the cuvette 101 contains a solid phase carrier 102 on which a probe for detecting a biological substance is immobilized, and a sample solution that is a liquid containing the biological substance. And a flow path 103.

  The solid phase carrier 102 is composed of a porous body and can pass through the sample solution. Although not limited to this, the solid-phase carrier 102 is, for example, a DNA chip made by spotting an oligo DNA probe on a porous substrate and immobilizing it.

  The channel 103 has a large-diameter portion 103a that extends downward from the upper surface opening across the solid-phase carrier 102, and a small-diameter portion 103b that extends laterally from the large-diameter portion 103a and terminates at the side surface. The large-diameter portion 103a is not limited to this, but has a truncated cone shape, for example. Although not limited to this, the small diameter part 103b has a taper, for example.

  The inspection apparatus includes a temperature adjustment mechanism 111 for adjusting the temperature of the attached inspection container 101, an observation window 112 that enables optical observation of the solid phase carrier 102 in the inspection container 101, and the observation window 112. An observation device 150 for acquiring an optical image of the solid phase carrier 102 in the cuvette 101 from above, a tube 114 fluidly connected to the small-diameter portion 103b of the flow path 103 of the cuvette 101 mounted, A pump 115 is fluidly connected to the tube 114. Among these, the temperature control mechanism 111 and the observation window 112 constitute an inspection stage that accommodates and holds the inspection container 101. The tube 114 and the pump 115 constitute a sample solution transfer mechanism that transfers the sample solution. Although the observation apparatus 150 is not limited to this, For example, it is comprised with optical microscopes, such as a fluorescence microscope.

  The observation window 112 has two transparent plates 113a and 113b arranged near the opening on the upper surface of the cuvette 101. The two transparent plates 113a and 113b are made of, for example, a thin glass plate, and are arranged substantially in parallel with an interval.

  The inspection stage including the temperature adjustment mechanism 111 includes a flow path 123 communicating with the space 125 between the two transparent plates 113a and 113b, a pump 120 for circulating a transparent liquid in the flow path 123, and a pump. 120 and a tube 121 that fluidly communicates with one end of the flow path 123, and a tube 122 that fluidly communicates the pump 120 and the other end of the flow path 123. The flow path 123 includes a heat exchange unit 124 for heating the circulating liquid by the heat generated by the temperature adjustment mechanism 111.

  The pump 120, the tubes 121 and 122, and the flow path 123 constitute an observation window heating unit that directly heats the observation window 112. This observation window heating means supplies the high-temperature transparent liquid heated by the heat exchange unit 124 to the space 125 between the transparent plates 113a and 113b constituting the observation window 112, and directly to the transparent plates 113a and 113b. The observation window 112 is heated by the contact.

  In the inspection stage, for example, as shown in FIG. 3, the inspection container and the observation window are configured separately. Here, the configuration of the inspection stage will be described in more detail with reference to FIG.

  As shown in FIG. 3, the inspection stage 130 that detachably holds the inspection container 101 has a base 131 having a placement part 131 a on which the inspection container 101 is placed and a receiving part 131 b against which the inspection container 101 is pressed. A cover member 132 that can be opened and closed with respect to the base 131, and a slider 133 for pressing the cuvette 101 against the receiving portion 131 b of the base 131.

  Although not shown, the cover member 132 is attached to, for example, an arm that is rotatably supported by a base on which the base 131 is placed, and can thereby be opened and closed with respect to the base 131. Here, the inspection stage 130 in which the cover member 132 is opened and closed with respect to the base 131 by rotation is illustrated, but the inspection stage 130 has a configuration in which the cover member 132 is opened and closed with respect to the base 131 by translation. May be.

  As indicated by the arrow, the slider 133 can move on the placement portion 131a of the base 131 in a direction approaching the receiving portion 131b and, conversely, a direction moving away from the receiving portion 131b.

  If the slider 133 is moved in a direction approaching the receiving portion 131b in a state where the cuvette 101 is placed on the mounting portion 131a of the base 131, the slider 133 comes into contact with the cuvette 101 and is then shown in FIG. As shown, the cuvette 101 is pressed against the side surface of the receiving portion 131 b of the base 131. As a result, the cuvette 101 is held.

  When the slider 133 is moved away from the receiving portion 131b from the state in which the cuvette 101 is held, the slider 133 moves away from the cuvette 101. In this state, the cuvette 101 can be easily removed from the mounting portion 131a of the base 131.

  The receiving portion 131b of the base 131 has a through hole 134 that crosses the side surface facing the side surface 104 where the flow path 103 of the cuvette 101 terminates. In a state where the inspection container 101 is held on the inspection stage 130, the inspection container 101 is hermetically coupled to the base 131 via the O-ring 106, and the flow path 103 of the inspection container 101 and the through hole 134 of the base 131 are fluidic. To be contacted. Thereby, the flow path 103 of the cuvette 101 is fluidly connected to the pump 115 via the through hole 134 of the base 131 and the tube 114.

  The inspection stage 130 includes a part of the components of the temperature adjustment mechanism 111 for controlling the temperature of the inspection container 101. The base 131 includes a heater 141 that generates heat in response to current supply and a temperature sensor 142 that detects the temperature of the heater 141. Further, the cover member 132 includes therein a heater 143 that generates heat in response to current supply and a temperature sensor 144 that detects the temperature of the heater 143.

  The cover member 132 is provided with two transparent plates 113a and 113b that are disposed substantially in parallel with each other and a flow path 123 that communicates with a space 125 between the two transparent plates 113a and 113b (FIG. 1). And FIG. 2). In FIG. 3, reference numeral 123 indicating the flow path is not shown, but here, FIG. 1 and FIG. 2 are inherited, and the flow path is denoted by reference numeral 123.

  As shown in FIG. 4, the flow path 123 includes a suction part 126 for taking in the transparent liquid supplied from the pump 120, a heat exchange part 124 for heating the transparent liquid, and discharging the transparent liquid. It has the discharge part 127 for doing.

  Although the suction part 126 and the discharge part 127 are not limited to this, For example, it opens in the side surface of the cover member 132. FIG. Tubes 121 and 122 (see FIG. 1) are connected to the suction part 126 and the discharge part 127, respectively.

  The heat exchanging portion 124 extends in a meandering manner in the portion where the heater 143 exists so that the liquid passing through the heat exchanging portion 124 is heated satisfactorily. One end of the heat exchange unit 124 is continuous with the suction unit 126, and the other end is continuous with the space 125 between the two transparent plates 113a and 113b. The discharge part 127 is continuous with the space 125 between the two transparent plates 113a and 113b.

  In the following description, it is assumed that the solid-phase carrier 102 is a DNA chip, and that the inspection device is a DNA chip analysis device and performs gene expression analysis.

  In the inspection container 101, a required amount of the sample solution 105 is dropped on the DNA chip 102 and mounted on the inspection apparatus (that is, accommodated and held on the inspection stage). The cuvette 101 containing the sample solution 105 is heated to an appropriate temperature of about 35 ° C. to 80 ° C. by the temperature adjustment mechanism 111 or controlled at an appropriate temperature cycle in order to promote the hybridization reaction.

  Further, a transparent liquid such as water is circulated through the flow path 123 provided in the temperature adjustment mechanism 111 (more precisely, the cover member 132) by the pump 120. The transparent liquid is heated by the heat exchange unit 124 and supplied to the space 125 between the two transparent plates 113a and 113b. As a result, the two transparent plates 113a and 113b are directly heated by the heated liquid.

  Further, in order to promote the hybridization reaction, the pump 115 pressurizes and depressurizes the flow path 103 in the cuvette 101 to force the sample solution 105 to pass through the DNA chip 102. As a result, the sample solution 105 is in a state where the liquid level is located on the DNA chip 102 as shown in FIG. 1 and in a state where the liquid level is located below the DNA chip 102 as shown in FIG. And repeatedly.

  By forcibly passing the DNA chip 102 through the sample solution 105 in this manner, the hybridization reaction can be completed within a relatively short time. For example, the hybridization reaction can be completed in about 1 to 2 hours.

  After completion of the reaction, the DNA chip 102 is observed for fluorescence by the observation device 150, and the increase or decrease in the expression level of the gene is read as the intensity of the fluorescence intensity.

  The inspection container 101 is open above the DNA chip 102, and the inspection stage has an observation window 112 above the DNA chip 102. For this reason, in such an inspection container 101 and an inspection stage, the DNA chip 102 is optically observed from above (normal microscope observation or the like) at any time before the reaction of the hybridization reaction, during the reaction, or after the completion of the reaction. Fluorescence observation). Therefore, the analysis can be performed by reading the increase or decrease in the expression level of the gene as the intensity of the fluorescence intensity.

  As described above, during the hybridization reaction, the cuvette 101 is warmed to a predetermined temperature by the temperature adjustment mechanism 111 or changed in a predetermined temperature cycle. At this time, in the present embodiment, the transparent liquid heated by the heat exchange unit 124 is always supplied to the space 125 between the two transparent plates 113a and 113b. The heated transparent liquid directly touches the transparent plates 113a and 113b, thereby heating the entire transparent plates 113a and 113b evenly. Therefore, the temperature distribution around the observation window 112 is as shown in FIG.

  As a comparative example, FIG. 10 shows the temperature distribution of the peripheral portion of the observation window in a configuration without the observation window heating means as in this embodiment (a configuration in which the observation window 112 is simply made of a single transparent plate).

  As can be seen by comparing FIG. 8 and FIG. 10, in the present embodiment, the temperatures of the transparent plates 113a and 113b are increased. For this reason, the heat radiation from the transparent plates 113a and 113b is satisfactorily suppressed. As a result, the temperature drop of the sample liquid can be suppressed, so that stable analysis can be performed.

  The transparent liquid supplied to the space 125 between the two transparent plates 113a and 113b does not hinder optical observation through the observation window 112.

Second Embodiment This embodiment is directed to another inspection apparatus for biologically related substances. FIG. 5 is a schematic cross-sectional view of the inspection apparatus and the inspection container attached thereto in the second embodiment of the present invention. 5, members denoted by the same reference numerals as those in FIG. 1 are equivalent members, and detailed description thereof is omitted here. In the following, the difference from the first embodiment will be described with emphasis.

  In the present embodiment, the observation window 112 includes a single transparent plate 160 and a heater 161 provided on the transparent plate 160. The inspection stage also has a control unit 162 that supplies current to the heater 161 and controls the amount of heat generated by the heater 161 by controlling the amount of current supplied. The heater 161 and the control unit 162 constitute observation window heating means for directly heating the observation window 112.

  Although the transparent plate 160 is not limited to this, For example, it is comprised with glass. In that case, in order to improve thermal conductivity, it is desirable that the thickness of the transparent plate 160 be relatively thin (about 0.1 to 1 mm).

  The heater 161 is formed of a heating resistor having an appropriate resistance that generates heat in response to current supply. The heating resistor is not limited to this, but may be composed of, for example, a metal film. In that case, the heater 161 is provided avoiding the region located above the DNA chip 102 of the cuvette 101.

  For example, as shown in FIG. 6, the heater 161 includes a ring-shaped conductive portion 163 that surrounds the DNA chip 102 of the cuvette 101 and a wiring portion 164 for supplying current that extends from the conductive portion 163. The

  For the cuvette 101 having a plurality of DNA chips 102, the heater 161 includes a ladder-shaped conductive portion 165 and a conductive portion 165 surrounding the plurality of DNA chips 102 as shown in FIG. And a wiring portion 166 for supplying current that extends from the wiring portion 166.

  In the present embodiment, the heater 161 provided directly on the transparent plate 160 generates heat in response to current supply from the control unit 162 to heat the transparent plate 160. As a result, the temperature distribution around the transparent plate 160 is as shown in FIG. As can be seen by comparing FIG. 9 and FIG. 10, in this embodiment, the temperature of the transparent plate 160 is increased. For this reason, the heat radiation from the transparent plate 160 is suppressed. As a result, the temperature drop of the sample liquid can be suppressed, so that stable analysis can be performed.

  So far, the heater 161 has been described assuming that it is made of an opaque conductive film, but the heater 161 may be made of a transparent conductive film. Although a transparent conductive film is not limited to this, For example, it is comprised with transparent conductive films, such as a tin dope indium oxide (ITO) film | membrane and a fluorine dope tin oxide (FTO) film | membrane.

  When the heater 161 is composed of a transparent conductive film, the heater (transparent conductive film) 161 may be provided on the entire top surface of the transparent plate 160. The transparent conductive film may be partially provided on the upper surface of the transparent plate 160 in a range including a region located above the DNA chip 102 of the test container 101 mounted on the test apparatus. As a result, the entire transparent plate 160 or a desired region (region located above the DNA chip 102) can be heated almost uniformly. As a result, heat dissipation from the transparent plate 160 can be more effectively suppressed, and more stable analysis can be expected.

  Although the embodiments of the present invention have been described above with reference to the drawings, the present invention is not limited to these embodiments, and various modifications and changes can be made without departing from the scope of the present invention. May be.

  Further, although a DNA chip using a porous substrate has been described as an example, the present invention can also be applied to a DNA chip using a silicon wafer or a slide glass as a base material. Moreover, it is applicable not only to the detection of DNA but also to the use of detecting an immune reaction using, for example, an antigen or an antibody as a probe.

It is a fragmentary sectional view showing typically composition of an inspection device in a first embodiment of the present invention, and an inspection container with which it is equipped, and the liquid level of a sample solution is located on a solid phase carrier. Indicates the state. It is the fragmentary sectional view which has shown typically the structure of the same test | inspection apparatus and test container as FIG. 1, and has shown the state in which the liquid level of a sample solution is located under a solid-phase carrier. It is sectional drawing of the principal part of the test | inspection stage which hold | maintains the test container in the test | inspection apparatus typically shown by FIG. 1 and FIG. 2 so that attachment or detachment is possible. FIG. 4 is a cross-sectional view of the cover member taken along line IV-IV shown in FIG. 3. It is typical sectional drawing of the inspection apparatus in 2nd embodiment of this invention, and the inspection container with which this was mounted | worn. FIG. 6 shows an example of the form of the heater shown in FIG. 5. FIG. 6 shows another example of the form of the heater shown in FIG. The temperature distribution of the peripheral part of the observation window in the inspection apparatus of 1st embodiment of this invention is shown. The temperature distribution of the peripheral part of the observation window in the inspection apparatus of 2nd embodiment of this invention is shown. The temperature distribution of the peripheral part of the observation window in the normal inspection apparatus which does not have an observation window heating means is shown.

Explanation of symbols

DESCRIPTION OF SYMBOLS 101 ... Test container, 102 ... DNA chip, 103 ... Channel, 103a ... Large diameter part, 103b ... Small diameter part, 104 ... Side surface, 105 ... Sample solution, 106 ... O-ring, 111 ... Temperature control mechanism, 112 ... Observation window 113a, 113b ... transparent plate, 114 ... tube, 115 ... pump, 120 ... pump, 121,122 ... tube, 123 ... flow path, 124 ... heat exchange part, 125 ... space, 126 ... suction part, 127 ... discharge part , 130 ... inspection stage, 131 ... base, 131 a ... mounting part, 131 b ... receiving part, 132 ... cover member, 133 ... slider, 134 ... through hole, 141 ... heater, 142 ... temperature sensor, 143 ... heater, 144 ... Temperature sensor, 150 ... observation device, 160 ... transparent plate, 161 ... heater, 162 ... control unit, 163 ... conductive unit, 164 ... Line unit, 165 ... conductive portion, 166 ... wire section.

Claims (10)

A temperature control mechanism for adjusting the temperature of the cuvette containing the solid phase carrier on which the probe for detecting the biological substance is solidified, and
An observation window that enables optical observation of the solid support;
An observation device for acquiring an optical image of the solid phase carrier in the cuvette through the observation window;
An inspection apparatus for biologically related substances, comprising observation window heating means for directly heating the observation window. In Claim 1, the observation window has two transparent plates arranged at an interval, and the observation window heating means has a flow path communicating with the space between the two transparent plates, and the flow path. And a pump for circulating a transparent liquid, the flow path has a heat exchange part, and the heat exchange part heats the transparent liquid passing therethrough by heat generated by a temperature control mechanism. 2. The inspection apparatus according to claim 1, wherein the observation window includes a transparent plate, and the observation window heating means includes a heater provided on the transparent plate and a control unit that controls the heater. 4. The inspection apparatus according to claim 3, wherein the heater is made of a metal thin film, and the metal thin film is positioned away from the observation region of the solid phase carrier. 4. The inspection apparatus according to claim 3, wherein the heater is made of a transparent conductive film, and the transparent conductive film is located in a range on the transparent plate including a region located above the observation region of the solid phase carrier. A temperature control mechanism for adjusting the temperature of the cuvette containing the solid phase carrier on which the probe for detecting the biological substance is solidified, and
An observation window that enables optical observation of the solid support;
An inspection stage for biologically related substances, comprising observation window heating means for directly heating the observation window. In Claim 6, the observation window has two transparent plates arranged at intervals, and the observation window heating means has a flow path communicating with the space between the two transparent plates, and the flow path. And a pump for circulating the transparent liquid, the flow path has a heat exchanging portion, and the heat exchanging portion heats the transparent liquid passing therethrough by heat generated by a temperature control mechanism. 7. The inspection stage according to claim 6, wherein the observation window includes a transparent plate, and the observation window heating means includes a heater provided on the transparent plate and a control unit that controls the heater. 9. The inspection stage according to claim 8, wherein the heater is composed of a metal thin film, and the metal thin film is located avoiding the observation region of the solid phase carrier. In claim 8, the heater is composed of a transparent conductive film, the transparent conductive film is located in the range on the transparent plate including the region located above the observation region of the solid phase carrier,
Inspection stage.

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