JP4767806B2 - Tip holder - Google Patents

Description

  The present invention relates to a chip holder for holding a channel chip.

  Conventionally, as a channel chip, for example, there are a chemical microdevice described in Patent Document 1, a microchannel device described in Patent Document 2, and the like. The flow path chip described in Patent Documents 1 and 2 includes a substrate and an elastic member bonded onto the substrate. A fine channel is formed at the joint between the substrate and the elastic member, and the chip is supplied from the channel so that the sample solution can be supplied into the channel and discharged from the channel. A supply port (liquid introduction hole) and a discharge port (liquid discharge hole) extend to the surface.

In such a channel chip, a sample (sample liquid) as a measurement target is supplied into the channel through the supply port, and, for example, as described in Patent Document 1, a voltage is applied to the sample in the channel. It is possible to carry out an electrophoretic analysis by applying. Further, as described in Patent Document 2, a DNA probe is fixed in a flow path and used as a DNA chip or the like.
JP 2004-219199 A Japanese Patent Laid-Open No. 2003-121311

  By the way, in recent years, for example, functional analysis of a cell information transmission function at a single cell level has been demanded using a channel chip having a channel. When performing a functional analysis of a biological sample such as a cell, it is preferable to flow a perfusate (sample solution) as a culture solution in the flow path in order to maintain a state in which the biological sample is utilized for a longer time.

  However, for example, it is difficult to flow a perfusate (sample solution) through a flow path by connecting a sample liquid supply pipe or the like to the flow path chip itself as described in Patent Document 1 from the outside. Therefore, it is desired to develop a chip holder that holds the flow channel chip and can easily flow the sample liquid into the flow channel in the flow channel chip.

  In addition, in the microchannel device described in Patent Document 2, a convex joint having a hole communicating therewith is provided on the supply port and the discharge port, and a concave joint fitting portion that fits the convex portion is provided. By connecting the connection device, it is possible to supply the perfusate to the flow channel in the flow channel chip and to discharge the perfusate from the flow channel. However, since there is only a direct connection device on the flow channel chip, it is difficult to align the flow channel chip and the connection device, and as a result, it may be difficult to supply the perfusate to the flow channel chip. . Therefore, the development of the tip holder as described above is desired.

  The present invention has been made in view of the above circumstances, and an object thereof is to provide a chip holder that holds a flow channel chip and can more easily flow a sample liquid into a flow channel in the flow channel chip. To do.

  The chip holder according to the present invention holds a flow channel chip having a flow channel therein and a liquid introduction hole for introducing the sample liquid into the flow channel and a liquid discharge hole for discharging the sample liquid from the flow channel. (1) a base member on which a flow path chip is placed; (2) attached to the base member, presses the flow path chip, and waits for a sample liquid to be supplied to the liquid introduction hole A well member having a well; (3) a cover that is attached to the base member, presses the flow path chip, and is connected to the liquid discharge hole; and a second pipe that is inserted into the well. (4) The well member has a connecting member for connecting the well and the liquid introduction hole and allowing the sample liquid in the well to flow into the liquid introduction hole.

  The chip holder has three members, a base member, a well member, and a cover member. After the flow channel chip is placed on the base member, the well member and the cover member are attached to the base member, so that the flow channel The chip is pressed and held in the chip holder. Thus, when the well member and the cover member are attached to the base member, as the well member presses the flow path chip, the connecting member of the well member is connected to the liquid introduction hole, and the well and the liquid introduction hole And are connected. Thereby, the sample liquid waiting in the well can be supplied to the flow path through the liquid introduction hole. Further, as the cover member presses the flow path chip, the first pipe is connected to the liquid discharge hole, and the second pipe is inserted into the well. Then, the sample liquid in the flow path is discharged through the liquid discharge hole using the first pipe, and the sample liquid is supplied into the well using the second pipe, or the sample liquid in the well is supplied. As a result, the volume of the sample solution in the well can be controlled.

  Therefore, for example, by connecting a pump or the like to the first pipe and sucking the inside of the flow path through the first pipe and the liquid discharge hole, the sample liquid waiting in the well can flow into the flow path. It is. Further, for example, by connecting a pump or the like to the second pipe and injecting the sample solution into the well, or by sucking out the excess sample solution in the well and discharging it out of the well, the sample solution in the well It is possible to control so that the capacity of is set to a predetermined amount. Therefore, after mounting the flow channel chip on the base member, it is possible to perform measurement using the flow channel chip while flowing the sample solution into the flow channel by simply attaching the well member and the cover member. In particular, since the first pipe and the second pipe are provided on the cover member, troublesome piping such as tubes connected to each pipe can be concentrated on the cover member, so that the chip holder can be handled. It becomes simple. In the chip holder, since the sample liquid can be temporarily kept in the well of the well member, it is easy to adjust the concentration of the sample liquid flowing in the flow path.

  Moreover, in the chip holder according to the present invention, it is preferable that the well and the connecting member are located substantially immediately above the liquid introduction hole. In this configuration, even when bubbles or the like enter the liquid introduction hole or the like when the connection member and the liquid introduction hole are connected, the bubbles or the like are easily removed through the connection member and the well.

  In the chip holder according to the present invention, the cover member has a third pipe inserted into the well, and one of the second pipe and the third pipe supplies the sample liquid to the well. It is preferable that the other is an overflow pipe for sucking a part of the sample solution in the well. In this configuration, the sample liquid can be supplied from the outside of the chip holder into the well through the supply pipe, and part of the sample liquid in the well is discharged by the overflow pipe, so that the sample liquid overflows from the well. It is controlled to a predetermined amount without going out.

  Further, in the tip holder according to the present invention, the end opposite to the end connected to the liquid discharge hole in the first pipe protrudes from the cover member, and in the second and third pipes, It is preferable that the end opposite to the end inserted into the well protrudes from the cover member.

  In this case, since the end of the first pipe opposite to the side connected to the liquid discharge hole protrudes from the cover member, the sample liquid in the flow path can be discharged through the liquid discharge hole and the first pipe. become. Furthermore, since the end opposite to the end inserted into the well in the second and third pipes protrudes from the cover member, the supply of the sample liquid into the well and the supply of the sample liquid from the well Emission is easier.

  Furthermore, in the chip holder according to the present invention in which the cover member has the second pipe and the third pipe, it is preferable that a step portion is formed inside the well. As described above, one of the second pipe and the third pipe is an overflow pipe. When a part of the sample liquid in the well is discharged with this overflow pipe, the water level of the sample liquid tends not to be constant due to surface tension or the like. It is possible to make it constant. As a result, for example, when a stimulating drug or the like is injected into a sample solution waiting in the well, the concentration of the stimulating drug or the like can be easily adjusted.

  Furthermore, in the chip holder according to the present invention, at least one of the base member and the cover member is formed with a window portion at a position facing the observation region in the flow path, and the well member is located on the observation region. It is preferable that it is attached to the base member so as not to be positioned. In this configuration, since the observation area of the flow channel in the flow channel chip can be observed through the window portion, for example, optical measurement or the like is possible.

  Further, in the chip holder according to the present invention, provided in the base member, provided in at least one of the first positioning means for positioning the flow path chip, the base member and the well member, It is preferable to further include a second positioning means for determining the position of the well member, and a third positioning means for determining the position of the cover member with respect to the base member, provided on at least one of the base member and the cover member.

  Since the position of the channel chip relative to the base member is determined by the first positioning means, the position of the well member relative to the base member is determined by the second positioning means, and the position of the cover member is determined by the third positioning means. The connecting member can reliably connect the well and the liquid introduction hole, and further, the first pipe and the liquid discharge hole are connected.

  In addition, as a 1st positioning means, the recessed part substantially the same shape as a flow-path chip is illustrated. Further, examples of the second positioning means include a positioning pin provided in the base member and a through hole that fits into the pin formed in the well member. The third positioning means is also exemplified by the same as the second positioning means.

  Furthermore, in the chip holder according to the present invention, the channel chip has n channels (n is an integer of 2 or more), and the liquid introduction hole and the liquid discharge are respectively provided in the n channels. The well member has n wells and connecting members, the cover member has n first and second pipes, and each connecting member has a corresponding well. It is preferable that the liquid introduction hole is connected, each first pipe is connected to the corresponding liquid discharge hole, and each second pipe is inserted into the corresponding well.

  In this case, each of the n second pipes can be used to supply the sample solution into each well, or the sample solution in each well can be sucked to control the volume of the sample solution in each well. become. Since the n wells are connected to the corresponding liquid introduction holes via the connecting members, the sample liquid can be supplied to each flow path. In addition, since each of the n first pipes is connected to the corresponding liquid discharge hole, the sample liquid can be discharged from each flow path. Therefore, it is easy to flow the sample solution through n channels.

  According to the chip holder according to the present invention, it is possible to easily flow the sample solution into the flow path in the flow path chip while holding the flow path chip.

  Hereinafter, embodiments of a chip holder according to the present invention will be described with reference to the drawings. In the description of the drawings, the same elements are denoted by the same reference numerals, and redundant description is omitted. Further, the dimensional ratios and the like in the drawings do not necessarily match those described.

(First embodiment)
The chip holder 1A shown in FIGS. 1 to 4 includes a base member 10, a well member 20, and a cover member 30, and the chip holder 1A is for holding the flow path chip 40A. In the following description, the side on which the cover member 30 is located with respect to the base member 10 is referred to as the upper side.

  Here, first, the configuration of the channel chip 40A held by the chip holder 1A will be described with reference to FIG. 7 showing the configuration of the channel chip 40A. FIG. 7A is a plan view of the channel chip. FIG. 7B is a side view of the flow path chip. FIG. 7C is a cross-sectional view taken along the line VIIc-VIIc in FIG. FIG. 7C shows an enlarged part of the cross-sectional configuration of the flow path chip 40A.

  The flow path chip 40A is configured by bonding an elastic member 42 made of, for example, PDMA to an upper surface 41a of a glass plate 41 as a substrate, and has a plate shape. The flow path chip 40A has eight fine flow paths 43, for example, micro flow paths, at the joint between the glass plate 41 and the elastic member. As shown in FIG. 7A, the eight flow paths 43 are formed so as to have observed regions (predetermined regions) 43a extending substantially in parallel with each other. As shown in FIG. 7C, the cell S as the measurement target is attached to at least a part of the wall surface. A liquid introduction hole 44 and a liquid discharge hole 45 extend from both ends of each flow path 43 to the upper surface 42 b of the elastic member 42.

  Next, the chip holder 1 </ b> A that holds the flow path chip 40 </ b> A will be described with reference to FIGS. 1 to 6. FIG. 1 is an exploded perspective view of the chip holder 1A. FIG. 2 is a plan view of the chip holder 1A. FIG. 3 is a side sectional view of the chip holder 1A. In FIG. 3, for the convenience of explanation, the characteristic portion of the chip holder 1A appears. 1 to 3 show a state in which the chip holder 1A holds the flow channel chip 40A. 4 is an enlarged view of a region surrounded by a one-dot chain line in FIG. FIG. 5 is a plan view of the well member 20. 6 is a cross-sectional view taken along the line VI-VI in FIG.

  As shown in FIGS. 1 to 3, the chip holder 1 </ b> A has a base member 10 on which the flow path chip 40 </ b> A is placed. In the present embodiment, the longitudinal direction of the base member 10 is referred to as a lateral direction, and a direction substantially orthogonal to the longitudinal direction is also referred to as a longitudinal direction.

  The base member 10 has a plate shape and is made of, for example, aluminum and stainless steel. The base member 10 has a recess 11 in which the well member 20 and the cover member 30 are mounted, and a recess (first positioning means) 12 in which the channel chip 40A is mounted is formed on the bottom wall surface of the recess 11. . The shape of the recess 12 is substantially the same shape as the flow channel chip 40A, and the depth of the recess 12 is substantially the same as the thickness of the flow channel chip 40A. Therefore, when the channel chip 40A is mounted in the recess 12, the position of the channel chip 40A with respect to the base member 10 is reliably determined, and the upper surface of the channel chip 40A is substantially flush with the bottom wall surface of the recess 11. Will be located on top. In order to facilitate attachment / detachment of the channel chip 40 </ b> A to the recess 12, it is preferable that a part of the side wall surface of the recess 12 is notched outward and the notch 14 is formed. In addition, an opening 13 that is an observation window is formed in a portion of the recess 12 that faces the observation region 43 a of the flow path 43.

  Further, on the bottom wall surface of the recess 11, pins 15 and 16 for positioning the well member 20 and the cover member 30 are provided on both sides of the recess 12, and on the side where the pins 15 and 16 are respectively provided, Screw holes 17 and 18 for screwing the well member 20 and the cover member 30 are further formed.

  The well member 20 is disposed on the side portion 11a side of the concave portion 11, and the well member 20 has a through hole 22-1 that fits into the pin 15 of the base member 10 and a through hole 22-2 through which the screw 51 passes. The first plate portion 21 is formed. The length of the first plate portion 21 in the vertical direction is substantially the same as the length of the concave portion 11 in the vertical direction. Also, the lateral length of the first plate portion 21 is such that the end portion 21a is such that the first plate portion 21 covers the liquid introduction hole 44 while the observation of the observation region 43a using the opening 13 is not hindered. Has a length that is not located on the observed region 43a. The second plate portion 25 extending in the vertical direction is integrally provided on the eight liquid introduction holes 44 on the end portion 21 a side of the first plate portion 21. The first plate portion 21 and the second plate portion 25 are made of, for example, aluminum and stainless steel.

  As shown in FIGS. 1 and 3, the well member 20 has a well 26 for waiting for a perfusate (sample liquid) W to be perfused in the flow path 43 immediately above each liquid introduction hole 44 of the flow path chip 40 </ b> A. . The shapes of the eight wells 26 are substantially the same, and the capacity thereof is, for example, 1 ml.

  As shown in FIGS. 5 and 6, the well 26 has a hole portion 26 a extending from the upper surface 25 a of the second plate portion 25 to the inside of the first plate portion 21, and a part of the inner wall on the upper end side of the hole portion 26 a. It is comprised from the pipe insertion part 26b, 26c cut off outside. A supply pipe 37 (second pipe) and an overflow pipe 38 (third pipe), which will be described later, are inserted into the pipe insertion portions 26b and 26c, respectively. Further, since the depths of the pipe insertion portions 26b and 26c are shallower than the thickness of the second plate portion 25, each well 26 has a step portion 26d inside.

  A stainless steel connecting pipe (connecting member) 27 extends from the lower end of each well 26, in other words, from the lower end of the hole 26 a toward the lower surface of the well member 20. It slightly protrudes from the lower surface of the member 20. Therefore, the connecting pipe 27 communicating with each well 26 is connected to the liquid introducing hole 44 corresponding to each well 26, and the liquid introducing hole 44 corresponding to each well 26 is connected. As a result, when the perfusate (sample liquid) W is supplied into the well 26, the perfusate W flows into the flow path 43 through the connection pipe 27 and the liquid introduction hole 44. Further, since the lower end portion 27a of the connecting pipe 27 slightly protrudes from the lower surface of the well member 20, when the well member 20 is screwed to the base member 10, as shown in FIGS. Since the connecting pipe 27 and the liquid introduction hole 44 are connected while the elastic member 42 is elastically deformed, liquid leakage from the connection portion between the connection pipe 27 and the liquid introduction hole 44 and intrusion of bubbles and the like are suppressed. Yes.

  In order to efficiently introduce the perfusate W into the liquid introduction hole 44 through the connection pipe 27, the well 26 preferably has a tapered shape that tapers toward the connection pipe 27. Further, from the viewpoint of allowing a slight positional deviation, the lower end portion 27a of the connecting pipe 27 is preferably chamfered as shown in FIG.

  Here, the stainless steel connecting pipe 27 is a connecting member that connects the well 26 and the liquid introduction hole 44. However, the connecting member may be a packing, for example, and the material is not limited to stainless steel. It is also possible to use rubber.

  As shown in FIGS. 1 to 3, the cover member 30 is disposed on the side portion 11 b side of the recess 11, and passes the through hole 32-1 fitted to the pin 16 of the base member 10 and the screw 52. The plate-shaped first cover part 31 in which the through hole 32-2 is formed. The length of the first cover portion 31 in the vertical direction is substantially the same as the length of the concave portion 11 in the vertical direction. Further, the lateral length of the first cover part 31 is such that the end part 31 a is in contact with or close to the end part 21 a of the first plate part 21.

  Further, the first cover portion 31 is connected to each liquid discharge hole 45 and has a stainless steel discharge pipe 33 (first pipe) for discharging the perfusate W in the flow path 43 through the liquid discharge hole 45. Arranged immediately above each liquid discharge hole 45. Each discharge pipe 33 extends in the thickness direction of the first cover portion 31 and is provided so as to penetrate the first cover portion 31. As shown in FIGS. 1 and 3, a concave portion is formed on the upper surface side of the portion where each discharge pipe 33 is disposed in the first cover portion 31 so that the upper end portion 33 a side of the discharge pipe 33 is exposed. It has become. Further, the lower end portion 33 b of the discharge pipe 33 slightly protrudes from the lower surface of the first cover portion 31 as in the case of the connection pipe 27. Thereby, when the cover member 30 is screwed to the base member 10, the elastic member 42 is elastically deformed and connected to the liquid discharge hole 45, so that at the connection portion between the discharge pipe 33 and the liquid discharge hole 45. Liquid leakage is suppressed.

  A plate-like second cover portion 34 is integrally provided at the end portion 31 a of the first cover portion 31. In addition, the 1st cover part 31 and the 2nd cover part 34 consist of aluminum and stainless steel, for example. Since the second cover part 34 is provided with respect to the first cover part 31 so as to protrude laterally from the upper side of the end part 31a, the first cover part 31 and the second cover part 34 are stepped. ing.

  On the lower surface side of the second cover portion 34, a recess 35 having substantially the same shape as the second plate portion 25 is formed so as to receive the second plate portion 25. An opening 36 is formed in the second cover portion 34 immediately above each hole portion 26a for injecting a stimulating drug into each well 26 using, for example, a dispensing head. Further, a stainless steel supply pipe 37 and an overflow pipe 38 are provided immediately above the pipe insertion portions 26b and 26c. The supply pipe 37 and the overflow pipe 38 extend in the thickness direction of the second cover portion 34 and are disposed so as to penetrate the second cover portion 34. The supply pipe 37 and the overflow pipe 38 have upper end portions 37a and 38a and lower end portions 37b and 38b protruding from the second cover portion 34. The lower end portion 37b of the supply pipe 37 is more than the lower end portion 38b of the overflow pipe 38. It is provided so as to be inserted into the well 26 deeply.

  In addition, an opening 39 is formed at a position directly above the opening 13 of the base member 10 and facing the observation region 43 a of the flow path 43. In the cover member 30 shown in FIGS. 1 to 3, a circular opening 39 is formed so as to cover a boundary portion between the first cover portion 31 and the second cover portion 34.

  When the flow path chip 40A is held by the chip holder 1A having the above configuration, first, the flow path chip 40A is mounted in the recess 12 of the base member 10. Since the recess 12 has substantially the same shape as the channel chip 40 </ b> A, the position of the channel chip 40 </ b> A is reliably determined with respect to the base member 10. Further, since the depth of the recess 12 and the thickness of the flow path chip 40A are substantially the same, the bottom wall surface of the recess 11 and the upper surface of the flow path chip 40A are located on substantially the same plane.

  Next, after fitting the pin 15 into the through hole 22-1, the well member 20 is screwed using the screw 51. Thereby, the channel chip 40 </ b> A is pressed by the first plate portion 21. As a result, since the connecting pipe 27 is pressed against the liquid introduction hole 44, the connection pipe 27 and the liquid introduction hole 44 are connected, and as a result, each well 26 and the corresponding liquid introduction hole 44 are connected. .

  Subsequently, after the pin 16 is fitted into the through hole 32-1, the cover member 30 is screwed to the base member 10 with a screw 52. Thereby, the channel chip 40 </ b> A is pressed by the first cover portion 31. As a result, since the discharge pipe 33 is pressed against the liquid discharge hole 45, the discharge pipe 33 and the liquid discharge hole 45 are connected. Further, the second plate portion 25 is accommodated in the concave portion 35 of the second cover portion 34, and the supply pipe 37 and the overflow pipe 38 are inserted into the corresponding pipe insertion portions 26b and 26c. When the cover member 30 is attached to the base member 10, the upper surface 25 a of the second plate portion 25 is in contact with the second cover portion 34, so that the second plate portion 25 is pressed by the second cover portion 34. By the pressing of the cover member 30 against the second plate portion 25, the connecting pipe 27 is further pressed toward the liquid introduction hole 44 side. As a result, it is possible to further prevent liquid leakage at the connecting portion and invasion of bubbles and the like.

  The tip holder 1A can be suitably used in the perfusion system 60 shown in FIG. The perfusion system 60 puts a stimulating agent as a reagent into the channel 43 at an arbitrary timing while irrigating the perfusate W as a culture solution into the channel chip 40A held by the chip holder 1A. This is for observing and measuring the reaction of the cells S in 43 to the stimulating drug. Here, a fluorescence probe is attached to the cell S in advance, and the fluorescence intensity is detected.

  As shown in FIG. 8, the perfusion system 60 includes a perfusion liquid tank 61, a waste liquid tank 62, an XY stage 63, an observation optical system 64, a dispensing head 65, and a dispensing head driving means 68. FIG. 8 shows a state in which the cells S in the flow path chip 40A held by the chip holder 1A are observed and measured.

  In the perfusate tank 61, a perfusate (sample solution) W as a culture solution to be passed through the flow path 43 is stored. The waste liquid tank 62 stores the perfusate W discharged from the flow path 43 through the liquid discharge hole 45. The XY stage 63 is for mounting the chip holder 1A holding the flow channel chip 40A, and has an opening 63a for observing the flow channel chip 40A from below.

  The observation optical system 64 is for optically observing and measuring the reaction of the cells S attached in the flow path 43 to the stimulating drug. The observation optical system 64 includes a light source unit 64A that outputs excitation light 71 having a wavelength that can excite the fluorescent probe attached to the cell S, a detection unit 64B such as a camera that detects fluorescence 72 from the fluorescent probe, and a light source. A light guide optical system 64C that irradiates the excitation light 71 output from the section 64A as the convergent light to the flow path 43 and condenses the fluorescence 72 from the fluorescent probe and inputs it to the detection section 64B. The light guide optical system 64C may include, for example, an objective lens and a dichroic mirror that transmits one of excitation light and fluorescence and reflects the other.

  The dispensing head 65 is for injecting stimulating drugs into the eight wells 26 at once, and a so-called eight-series dispensing head is exemplified. The dispensing head driving means 68 moves the dispensing head 65 three-dimensionally and controls injection of the stimulating drug into the dispensing head 65 and discharge of the stimulating drug.

  Next, a method for using the tip holder 1A in the perfusion system 60 will be described.

  First, a channel chip 40A in which cells S to which a fluorescent probe is attached is seeded and cultured in the channel 43 and a chip holder 1A are prepared. At this time, the chip holder 1 </ b> A is disassembled into the base member 10, the well member 20, and the cover member 30.

  Then, the supply pipe 37 of the cover member 30 and the perfusate tank 61 are connected by a tube 66a, and the discharge pipe 33 and overflow pipe 38 of the cover member 30 and the waste liquid tank 62 are connected by a tube 66b and a tube 66c. The member 30 is placed above the observation position.

  Next, the channel chip 40A is attached to the recess 12 of the base member 10 of the chip holder 1A, and the well member 20 is attached to the base member 10. Thereafter, the base member 10 on which the flow path chip 40 </ b> A and the well member 20 are mounted is set on the XY stage 63. At this time, the base member 10 is set so that the opening 13 is positioned on the opening 63 a of the XY stage 63. Subsequently, after the XY stage 63 is driven and moved to a predetermined observation position on the observation optical system 64 as shown in FIG. 8, the cover member 30 is screwed to the base member 10 and the chip holder 1A and To do.

  Next, the pump 67a provided on the tube 66a is driven from the perfusate tank 61 to deliver the perfusate W, while the pump 67b provided on the tube 66b is driven to pass through the tube 66b and the discharge pipe 33. The inside of the flow path 43 is sucked.

  The perfusate W delivered from the perfusate tank 61 by the pump 67 a flows into the well 26 through the supply pipe 37, and then flows from the lower end of the well 26 through the connection pipe 27 and the liquid introduction hole 44. Flows in. Since the flow path 43 is sucked, the perfusate W flowing into the flow path 43 is discharged to the waste liquid tank 62 through the liquid discharge hole 45, the discharge pipe 33, and the tube 66b. Accordingly, the perfusate W perfuses the flow path 43.

  When the perfusate W is introduced into the well 26, in order to prevent the perfusate W from overflowing from the well 26, the pump 67c provided on the tube 66c is driven, and the tube 66b and the overflow pipe 38 are connected. Through the well 26, the perfusate W above a certain water level is discharged to the waste liquid tank 62.

  When perfusion of the perfusate W into the flow path 43 is started, the excitation light 71 is output from the light source unit 64A of the observation optical system 64, and the excitation light 71 is opened by the light guide optical system 64C. 13 and the glass plate 41 irradiate the cells S in the observation region 43a in the eight flow paths 43 as convergent light. Since the fluorescent probe is attached to the cell S, the fluorescence 72 is generated by the irradiation of the excitation light 71. The generated fluorescence 72 is collected by the light guide optical system 64C and input to the detection unit 64B to be detected. Thereby, the cells S in the observed region 43a are observed.

  Next, the dispensing head driving means 68 causes the dispensing head 65 to access a ligand plate (not shown) holding the stimulating drug, and causes the dispensing head 65 to suck the stimulating drug from the ligand plate. Then, as shown in FIG. 8, a part of the dispensing head 65 is inserted into the well 26 through the opening 36 by the dispensing head driving means 68, and the stimulating drug is injected into the perfusate W in the well 26. When the perfusate W into which the stimulating drug is injected flows into the flow path 43, the cells S react with the stimulating drug, and the fluorescence intensity changes. Therefore, the response of the cell S to the stimulating drug is measured by detecting the change in the fluorescence intensity due to the administration of the stimulating drug by the detection unit 64B.

  Next, functions and effects of the chip holder 1A will be described.

  In the chip holder 1 </ b> A, the flow path chip 40 </ b> A is fixed to the base member 10 at a fixed position because the flow path chip 40 </ b> A is attached to the recess 12 having substantially the same shape as the flow path chip 40 </ b> A. Moreover, the well member 20 and the cover member 30 are fitted to the pins 15 and 16 provided on the base member 10 by fitting the through holes 22-1 and the through holes 32-1 of the well member 20 and the cover member 30, respectively. Since the alignment is performed, the well member 20 and the cover member 30 are securely fixed to a certain position of the base member 10. Therefore, even if the well member 20 and the cover member 30 are attached to the base member 10 after the flow channel chip 40A is attached to the base member 10, the connecting pipe 27 and the liquid introduction hole 44 are securely connected and discharged. The pipe 33 and the liquid discharge hole 45 are securely connected.

  Therefore, as described using the perfusion system 60, the flow channel chip 40A is placed on the base member 10, and after the well member 20 is mounted on the base member 10, it is transported to a predetermined observation position and pre-tube 66a. The analysis for elucidating the function of the cell S can be easily performed simply by mounting the cover member 30 to which ~ 66c is connected to the base member 10, and for example, the measurement can be automated.

  Further, since the chip holder 1 </ b> A has the wells 26 corresponding to the number of the flow paths 43, the stimulating drug can be collectively dispensed into the eight wells 26. As a result, collective measurement in the eight flow paths 43 is possible, and a large amount of measurement of the response of the cells S to the stimulating drug can be performed.

  Further, since the connection pipe 27 is pressed and connected to the liquid introduction hole 44, liquid leakage at the connection portion between the connection pipe 27 and the liquid introduction hole 44 and intrusion of bubbles or the like into the liquid introduction hole 44 are suppressed. ing. As described above, intrusion of bubbles or the like into the liquid introduction hole 44 is suppressed. Even if bubbles or the like enter the liquid introduction hole 44 at the time of connection, the connection pipe 27 and the well are directly above the liquid introduction hole 44. Since 26 is located, the bubbles rise in the connecting pipe 27 and the well 26 and are discharged. Furthermore, the path from the well 26 to the liquid introduction hole 44 is shorter due to the above-described arrangement relationship of the liquid introduction hole 44, the connecting pipe 27, and the well 26. The drug easily flows into the flow path 43 quickly. From the viewpoint of measuring the reaction of the cells S in each channel 43 due to the injection of the stimulating drug into each well 26 at approximately the same timing, the path length from the lower end of each well 43 to the liquid introduction hole 44 is the same. Is preferred.

  Further, in the tip holder 1A, the perfusate W is made to wait in the well 26 of the well member 20, and the stimulating drug is put into the perfusate W in the well 26 using a dispensing head or the like. Easy to control.

  Further, since the step portion 26d is provided in the well 26, when the perfusate W is introduced into the well 26, the liquid level of the perfusate W becomes constant. If no step is formed in the well, when a part of the perfusate W in the well 26 is sucked using the overflow pipe 38, it is transferred to the overflow pipe 38 along the inner wall of the well 26 due to surface tension or the like. There is a possibility that the perfusate W flows in and the liquid level does not become constant. Thus, when the water level of the perfusate W in the well 26 is not constant, it is difficult to control the concentration when the stimulating drug is injected. On the other hand, in the chip holder 1A, since the step portion 26d is provided, the liquid level in the well 26 becomes constant, and as a result, the concentration of the stimulating drug can be further controlled.

  In the present embodiment, a part of the edge portion on the upper end side of the hole portion 26a is notched to form pipe insertion portions 26b and 26c shallower than the depth of the hole portion 26a. Although the step portion 26d is provided, for example, the step portion 26d may be formed over the entire circumference of the hole portion 26a.

  Further, since the liquid level tends to be constant as described above by having the step portion 26d, the step portion 26d is preferably formed in the well 26, but the step portion 26d is not formed. Also good. Further, in order to adjust the amount of the perfusate W in the well 26, an overflow pipe 38 is provided. However, the amount of the perfusate W introduced into the well 26 and the amount of the perfusate W discharged from the flow path 43 are described. In this case, the overflow pipe 38 may not be provided. Furthermore, in order to adjust the amount of the perfusate W in the well 26, in the above embodiment, the two pipes of the supply pipe 37 and the overflow pipe 38 are exemplified, but the present invention is not limited to this. For example, if a pump capable of sucking and discharging is used, the amount of liquid can be adjusted with a single pipe.

(Second Embodiment)
Next, a second embodiment of the chip holder according to the present invention will be described with reference to FIGS.

  As shown in FIGS. 9 and 10, the chip holder 1B is for holding the flow path chip 40B.

  Also in this embodiment, the flow channel chip 40B will be described first with reference to FIG. In the configuration of the channel chip 40B, in the eight channels 43, the distance from the liquid introduction hole 44 to the observation region 43a is substantially the same, and the distance from the liquid discharge hole 45 to the observation region 43a is substantially the same. Thus, it differs from the configuration of the channel chip 40A in that the channel 43 is formed. In this configuration, the liquid introduction hole 44 and the liquid discharge hole 45 are arranged in an arc as shown in FIG.

  Next, the chip holder 1B that holds the flow path chip 40B will be described. The chip holder 1 </ b> B is configured by attaching a well plate (well member) 90 and a perfusion plate (cover member) 100 to a base member 80. The materials of the base member 80, the well plate 90, and the perfusion plate 100 can be the same as the materials of the base member 10, the well member 20, and the cover member 30 exemplified in the first embodiment.

  The base member 80 has a substantially rectangular shape when viewed from above, and has a recess 11 as in the case of the base member 10, and further has a recess 12 having substantially the same shape as the channel chip 40 </ b> B in the recess 11. Is formed. An opening 13 as an observation window is formed at the center of the recess 12. Further, a pin 16 for positioning the perfusion plate 100 and a screw hole 18 for receiving a screw 52 for screwing the perfusion plate 100 are formed on the bottom wall surface of the recess 11.

  Also in this embodiment, the longitudinal direction of the base member 80 is referred to as a lateral direction, and a direction substantially orthogonal to the longitudinal direction is also referred to as a longitudinal direction. In the present embodiment, the “upper side” is the side on which the perfusion plate 100 is disposed with respect to the base member 80.

  The well plate 90 is placed on the base member 80 so that the end portion 90 a is in contact with the side portion 11 a of the recess 11. As shown in FIG. 9, the length of the well plate 90 in the vertical direction is substantially the same as the length of the recess 11 in the vertical direction, so that the well plate 90 is placed on the base member 80 as described above. The position is determined. Therefore, the recess 11 functions as a positioning means (second positioning means) for the well plate 90.

  As shown in FIG. 11, the well plate 90 has eight wells 91 of the same size on the end 90a side. Each well 91 has a substantially rectangular shape when viewed from above, and a groove 92 extends from each well 91 toward the end 90b to a position directly above the liquid introduction hole 44. A connecting pipe 27 connected to the liquid introduction hole 44 extends from the lower part of the end 92 a of each groove 92 toward the lower surface of the well plate 90, and the lower end of the connecting pipe 27 is slightly from the lower surface of the well plate 90. Protruding.

  As shown in FIGS. 9 and 10, the perfusion plate 100 is such that a part of the lower surface on the end 100 a side can cover the groove 92 of the well plate 90 and the connection side of the well 91 to the groove 92. Notched into. The perfusion plate 100 covers the flow path chip 40 </ b> B and covers the end 90 b side of the well plate 90 so that a part of the well 91 is exposed in the well plate 90.

  The perfusion plate 100 is formed with a through hole 32-1 that fits into the pin 16 of the base member 80 and a through hole 32-2 through which the screw 52 passes. Further, an opening 39 is formed in a portion of the perfusion plate 100 immediately above the opening 13 of the base member 80 and facing the observation region 43 a of the flow path 43. Further, a discharge pipe 33 is disposed immediately above the liquid discharge hole 45 of the flow path chip 40B, as in the case of the first embodiment, and a supply pipe 37 and an overflow pipe are directly above each well 91. 38 is arranged in the same manner as in the first embodiment.

  In the chip holder 1 </ b> B configured as described above, the well plate 90 is mounted on the base member 80 as described above after the flow path chip 40 </ b> B is attached in the recess 12 of the base member 80. Next, the perfusion plate 100 is positioned using the pins 16 and the through holes 32-1, and then screwed.

  When the tip holder 1B is assembled in this way, the lower end portions 37b and 38b of the supply pipe 37 and the overflow pipe 38 are inserted into the portion of the well 91 covered by the perfusion plate 100. Further, when the perfusion plate 100 is screwed to the base member 80, the discharge pipe 33 is pressed against the liquid discharge hole 45 and the discharge pipe 33 and the liquid discharge hole 45 are connected. Furthermore, since the well plate 90 is pressed by the perfusion plate 100, the connecting pipe 27 is pressed by the liquid introduction hole 44. As a result, the connection pipe 27 and the liquid introduction hole 44 are connected, and the liquid introduction hole 44 and the well 26 are connected via the connection pipe 27 and the groove 92.

  The tip holder 1B is suitably used, for example, in the perfusion system 60 shown in FIG. 8 as in the case of the tip holder 1A of the first embodiment. The chip holder 1B is used in the perfusion system 60. The chip holder 1B is assembled by positioning and positioning the well plate 90 in contact with the side portion 11a of the recess 11, and then using the perfusion plate 100 with the well plate 90. Is the same as in the case of the tip holder 1A except that a part of the dispensing head 65 is inserted into a part of the well 91 exposed from the perfusion plate 100.

  The action and effect of the chip holder 1B is the same as that of the chip holder 1A. That is, also in the tip holder 1B, the positions of the flow path tip 40B, the well plate 90, and the perfusion plate 100 are determined with respect to the base member 80. Therefore, by assembling the tip holder 1B, the connection pipe 27 and the liquid introduction hole 44 are assembled. Are securely connected, and the discharge pipe 33 and the liquid discharge hole 45 are securely connected. Further, the perfusion plate 100 is screwed to the base member 80, whereby the well plate 90 is pressed and the connection pipe 27 and the liquid introduction hole 44 are connected. As a result, liquid leakage is prevented at the connection portion between the connection pipe 27 and the liquid introduction hole 44, and further, bubbles and the like are prevented from entering the flow path 43.

  Further, since the tip holder 1B waits for the perfusate W in the well 91 and supplies the stimulating drug to the perfusate W in the well 91 using a dispensing head or the like, the concentration control of the stimulating drug is easy. . From the viewpoint of further facilitating such concentration control, it is preferable to form a stepped portion inside the well 91. Further, from the viewpoint of measuring the reaction of the cells S in each flow path 43 due to the injection of the stimulating drug into each well 91, the path length from each well 91 to the corresponding liquid introduction hole 44 is the same. Preferably there is.

  In the tip holder 1B, the perfusion plate 100 as the cover member includes the supply pipe 37 and the overflow pipe 38. However, the perfusion plate 100 does not include either the supply pipe 37 or the overflow pipe 38. It is also possible. In this case, a supply pipe or an overflow pipe may be directly inserted into the portion of the well 91 that is not covered with the perfusion plate 100. Even in this case, for example, it is easier to supply the perfusate W to the flow path 43 than when a supply pipe or an overflow pipe is directly connected to the liquid introduction hole 44 of the flow path chip 40B. Furthermore, in order to adjust the amount of the perfusate W in the well 91, in the above embodiment, the two pipes of the supply pipe 37 and the overflow pipe 38 are exemplified, but the present invention is not limited to this. For example, if a pump capable of sucking and discharging is used, the amount of liquid can be adjusted with a single pipe.

  As mentioned above, although preferred embodiment of the chip holder of this invention was described, this invention is not limited to the said embodiment. In the first embodiment, the eight connecting pipes (connecting members) 27 are arranged on a straight line. For example, when the flow channel chip 40B shown in FIG. 12 is used, as shown in FIG. In addition, the arrangement state of the connecting pipes 27 may be arcuate. In the tip holder 1A of the first embodiment, when holding the flow channel tip 40B, the arrangement state of the discharge pipes 33 is made arcuate in accordance with the arrangement state of the liquid discharge holes 45 of the flow channel tip 40B. Keep it. Further, in the tip holder 1B of the second embodiment, when the flow channel tip 40A is held, the arrangement state of the connection pipe 27 provided in the well plate 90 and the discharge pipe 33 provided in the perfusion plate 100 is linear. You can do it.

  Furthermore, in the case of the first embodiment, the well 26 is positioned immediately above the liquid introduction hole 44, but the present invention is not limited to this. When the well 26 is not located immediately above the corresponding liquid introduction hole 44, a groove portion may be provided as in the second embodiment to connect the well 26 and the connection pipe 27, or a bent connection pipe. By using the (connection member) 27, the well 26 and the liquid introduction hole 44 can be connected. The supply pipe 37, the overflow pipe 38, and the discharge pipe 33 are linear, but may be bent.

  Further, the tip holders 1A and 1B have the openings 13 and 39 as window portions so that they can be used for optical observation and measurement by the observation optical system 64 of the perfusion system 60. When the measurement is not performed, for example, when the flow path chip as the DNA chip is held by the chip holders 1A and 1B, the openings 13 and 39 may not be provided.

  Even in the case of optical observation, it is sufficient that either one of the openings 13 and 39 is formed. In the case where the openings 13 and 39 are formed, the light that is not absorbed by the fluorescent probe among the excitation light incident through one of the openings 13 and 39 passes through the other opening and the chip holders 1A and 1B. This is preferable because it can be detected with higher sensitivity. Moreover, although the window part was made into opening 13 and 39, it is not limited to this, You may be comprised from the transparent member which can observe flow-path chip | tip 40A, 40B.

  Further, in the chip holder 1A, the number of the wells 26, the connecting pipes 27, the discharge pipes 33, the supply pipes 37, and the overflow pipes 38 need only be the same as the number of the flow paths 43 included in the flow path chip 40A. For example, when the number of the flow paths 43 is one, the number of the wells 26 may be one, and the number of the connecting pipes 27, the discharge pipes 33, the supply pipes 37, and the overflow pipes 38 may be one. When the number of the flow paths 43 is n (n is an integer of 2 or more), the number of the wells 26 may be n, and the connection pipe 27, the discharge pipe 33, the supply pipe 37, and the overflow pipe 38 are each n. I need a book. The same applies to the chip holder 1B.

  In addition, the lower end portion 27 a of the connecting member 27 protrudes from the lower surface of the well member 20, but it only needs to be connected to the liquid introduction hole 44. The same applies to the discharge pipe 33.

  Furthermore, although the first positioning means for positioning the channel chips 40A and 40B is the recess 12, it is not particularly limited as long as the channel chips 40A and 40B can be arranged at predetermined positions of the base member 10, for example, the channel chip 40A, A raised portion that defines the position of the corner of 40B may be formed on the base member 10. Furthermore, in the first embodiment, the second positioning means is composed of the pin 16 and the through hole 22-1; however, as shown in the second embodiment, the concave portion 11 can be used. Further, the third positioning means is not limited to the pin 16 and the through hole 32-1, and for example, the recess 11 can be used.

  Further, from the viewpoint of, for example, automating measurement using the perfusion system 60, it is preferable to include the first positioning unit, the second positioning unit, and the third positioning unit, but it is not always necessary. For example, after marking a predetermined position of the base member 10 and disposing the flow path chips 40A and 40B in accordance with the mark, the well members 20 and 90 and the cover members 30 and 100 are replaced with screws 51 and 52, for example. If the connection pipe 27 and the liquid introduction hole 44 are fixed, the discharge pipe 33 and the liquid discharge hole 45 can be connected to each other.

  Furthermore, in the chip holder 1A of the first embodiment, the well member 20 and the cover member 30 are screwed to the base member 10 using the screws 51 and 52, so that the well member 20 and the cover member 30 are connected to the channel chip. 40A and 40B are pressed. However, for example, when the perfusion system 60 includes a pressing unit that presses the well member 20 and the cover member 30, the fixing is not necessarily limited to screwing. The same applies to the chip holder 1B.

  Furthermore, although the upper surfaces of the channel chips 40A and 40B held by the chip holders 1A and 1B are planar, for example, as described in Patent Document 2, they communicate with the liquid introduction hole 44 and the liquid discharge hole 45. The convex part which has a hole part may be provided. In this case, a concave portion for receiving the convex portion provided on the flow path chips 40A and 40B may be formed at corresponding positions in the well members 20 and 90 and the cover members 30 and 100.

  The sample solution W is a perfusate W as a culture solution. However, as described above, when the flow channel chips 40A and 40B as DNA chips are used, a solution containing a sample (DNA) to be detected is used. do it.

It is a disassembled perspective view of one Embodiment of the chip holder which concerns on this invention. It is a top view of the chip holder shown in FIG. It is a sectional side view of the chip holder shown in FIG. It is an enlarged view of the area | region enclosed with the dashed-dotted line of FIG. It is a top view of the well member which the chip holder shown in FIG. 1 has. FIG. 6 is a cross-sectional view taken along line VI-VI in FIG. 5. It is a figure which shows the flow-path chip | tip which the chip | tip holder shown in FIG. 1 hold | maintains. It is a schematic block diagram of a perfusion system. It is a top view of other embodiments of a chip holder concerning the present invention. FIG. 10 is a side sectional view of the chip holder shown in FIG. 9. It is a top view of the well member which the chip holder shown in FIG. 9 has. FIG. 10 is a plan view of a flow path chip held by the chip holder illustrated in FIG. 9. FIG. 6 is a plan view of another example of the well member shown in FIG. 5.

Explanation of symbols

  DESCRIPTION OF SYMBOLS 1A, 1B ... Chip holder, 10 ... Base member, 11 ... Recessed part (2nd positioning means), 12 ... Recessed part (1st positioning means), 13, 39 ... Opening (window part), 15 ... Pin (2nd positioning means) , 16 ... Pin (third positioning means), 20 ... Well member, 22-1 ... Through hole (second positioning means), 26 ... Well, 26d ... Stepped portion, 27 ... Connection pipe (connection member), 30 ... Cover member, 32-1 ... through hole (third positioning means), 33 ... discharge pipe (first pipe), 33a ... upper end (end opposite to the side connected to the liquid discharge hole), 37 ... supply pipe (second pipe), 37a ... upper end (end opposite to the side inserted into the well), 38 ... overflow pipe (third pipe), 38a ... upper end (inserted into the well) End opposite side), 40A, 40B ... flow Chips 43... Channels 43 a. Observation region 44. Liquid introduction holes 45. Liquid discharge holes 80. Base members 90. Well plates (well members) 91 91 Wells 100 Perfusion plates (cover members) ), W ... perfusate (sample solution).

Claims (8)

A chip holder that holds a flow channel chip having a flow channel therein and having a liquid introduction hole for introducing a sample liquid into the flow channel and a liquid discharge hole for discharging the sample liquid from the flow channel. And
A base member on which the flow path chip is placed;
A well member attached to the base member and having a well for pressing the flow channel chip and waiting for the sample liquid to be supplied to the liquid introduction hole;
A cover member that is attached to the base member, presses the flow path chip, and has a first pipe connected to the liquid discharge hole, and a second pipe inserted into the well;
With
The well member includes a connection member that connects the well and the liquid introduction hole and allows the sample liquid in the well to flow into the liquid introduction hole.   The chip holder according to claim 1, wherein the well and the connecting member are positioned substantially immediately above the liquid introduction hole. The cover member has a third pipe inserted into the well,
One of the second pipe and the third pipe is a supply pipe for supplying the sample liquid to the well, and the other is for sucking a part of the sample liquid in the well The chip holder according to claim 1, wherein the chip holder is an overflow pipe. In the first pipe, an end opposite to the end connected to the liquid discharge hole protrudes from the cover member,
4. The chip holder according to claim 3, wherein, in the second and third pipes, an end opposite to an end inserted into the well protrudes from the cover member. 5.   The chip holder according to claim 3, wherein a step portion is formed inside the well. At least one of the base member and the cover member is formed with a window at a position facing the observation region in the flow path,
The chip holder according to claim 1, wherein the well member is attached to the base member so as not to be positioned on the observation region. A first positioning means provided on the base member for positioning the flow path chip;
Second positioning means provided on at least one of the base member and the well member, and determining a position of the well member with respect to the base member;
A third positioning means which is provided on at least one of the base member and the cover member and determines the position of the cover member with respect to the base member;
The chip holder according to any one of claims 1 to 6, further comprising: The channel chip has n channels (n is an integer of 2 or more), and the liquid introduction hole and the liquid discharge hole are formed for each of the n channels.
The well member has n wells and connection members,
The cover member has n pieces of the first and second pipes,
Each connection member connects the corresponding well and the liquid introduction hole,
Each of the first pipes is connected to the corresponding liquid discharge hole,
The chip holder according to claim 1, wherein each second pipe is inserted into the corresponding well.

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