JP2012185008A - Channel chip for cell assay and inspection apparatus - Google Patents

Channel chip for cell assay and inspection apparatus Download PDF

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JP2012185008A
JP2012185008A JP2011047678A JP2011047678A JP2012185008A JP 2012185008 A JP2012185008 A JP 2012185008A JP 2011047678 A JP2011047678 A JP 2011047678A JP 2011047678 A JP2011047678 A JP 2011047678A JP 2012185008 A JP2012185008 A JP 2012185008A
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cell assay
cell
chip
solution
channel chip
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JP5872776B2 (en
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Yukihiro Sugiyama
幸宏 杉山
Takao Okada
孝夫 岡田
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RES INST OF BIOMOLECULE METROLOGY CO Ltd
Research Institute of Biomolecule Metrology Co Ltd
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RES INST OF BIOMOLECULE METROLOGY CO Ltd
Research Institute of Biomolecule Metrology Co Ltd
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Abstract

PROBLEM TO BE SOLVED: To provide a channel chip for cell assay in which a cell can be cultivated without giving stress thereto, and an inspection apparatus therefor.SOLUTION: In a channel chip for cell assay capable of cell cultivation, an injection port 5 and a discharge port 7 for a solution are provided on an upper surface of a channel chip body 3 for cell assay. In a channel 11 communicating the injection port 5 and the discharge port 7, a dam section 21 is provided for damming back cells 19 in the solution injected from the injection port 5. A cultivation chamber 25 is provided at an upstream side of the dam section 21. At a downstream side of the dam section 21, in order to suppress a flow rate of the solution flowing through the dam section 21 to low rate, a liquid storage chamber 27 is provided which keeps a liquid level higher than the dam section during cell cultivation. A solution holding cup 17 for holding the solution at a predetermined height is provided in the injection port 5 and the discharge port 7. The capacity of the liquid storage chamber 27 is made less than that of the cultivation chamber 25, and at least one of upper and lower walls of the cultivation chamber 25 is transparent.

Description

本発明は、細胞培養が可能な細胞アッセイ用流路チップに係り、さらに詳細には、培養中の細胞に対するストレスの付与を抑制することの可能な細胞アッセイ用流路チップ、及び当該細胞アッセイ用流路チップを使用して細胞の生理活性を評価することが可能な検査装置に関する。   The present invention relates to a cell assay channel chip capable of culturing cells. More specifically, the present invention relates to a cell assay channel chip capable of suppressing the application of stress to cells in culture, and the cell assay. The present invention relates to an inspection apparatus capable of evaluating a physiological activity of a cell using a channel chip.

従来、細胞の生理活性を測定するには、先ず、例えば細胞接着性のプラスチック製ウエル（数１００μＬ程度）内に細胞を植付け、次に細胞と反応させる試薬の存在下で細胞を培養し、最後に発色／発光染色処理をして、光学的検出をすることにより、行っている（例えば、特許文献１）。   Conventionally, in order to measure the physiological activity of a cell, first, for example, the cell is implanted in a cell-adhesive plastic well (about several hundred μL), and then the cell is cultured in the presence of a reagent that reacts with the cell. This is performed by performing color development / luminescence staining treatment and optical detection (for example, Patent Document 1).

特開２００８−３９５４１号公報JP 2008-39541 A

ところで、前述したように、プラスチック製ウエル内に細胞を植付けてから細胞の生理活性を測定するまでの一連の工程において、培地の交換、試薬の注入、ウエル内の洗浄などの操作は、人の手によるピペット操作によって実施されている。したがって、操作工数が多く煩雑である。例えば、ピペット操作による溶液の交換時には、ウエル内の溶液を攪拌し、細胞を舞上げることがあり、培養中の細胞にストレスを付与することが多い、という問題がある。   By the way, as described above, in a series of steps from planting a cell into a plastic well to measuring the physiological activity of the cell, operations such as medium replacement, reagent injection, and well washing are performed by humans. It is carried out by hand pipetting. Therefore, the number of operation steps is large and complicated. For example, at the time of exchanging the solution by pipetting, there is a problem that the solution in the well may be agitated to soar the cells, and stress is often applied to the cells being cultured.

本発明は、前述のごとき問題に鑑みてなされたもので、細胞培養が可能な細胞アッセイ用流路チップであって、当該細胞アッセイ用流路チップ本体の上面に溶液の注入口と排出口とを備え、前記注入口と排出口とを連通した流路中に、前記注入口から注入された溶液内の細胞をせき止めするためのせき止め部を備え、当該せき止め部の上流側に培養室を備えると共に、前記せき止め部の下流側に、前記せき止め部を流れる溶液の流速を低速に抑制するために、細胞培養時には前記せき止め部よりも液面高さを高く保持する液溜室を備えていることを特徴とするものである。   The present invention has been made in view of the above-described problems, and is a cell assay channel chip capable of culturing cells, and a solution injection port and a discharge port on the upper surface of the cell assay channel chip body. And a damming portion for damaging cells in the solution injected from the inlet, and a culture chamber upstream of the damming portion. In addition, in order to suppress the flow rate of the solution flowing through the damming portion at a low speed on the downstream side of the damming portion, a liquid reservoir chamber that holds a higher liquid level than the damming portion during cell culture is provided. It is characterized by.

また、前記細胞アッセイ用流路チップにおいて、前記注入口及び排出口に、溶液を所定高さに保持する溶液保持カップを備えていることを特徴とするものである。   In the cell assay channel chip, the inlet and the outlet have a solution holding cup for holding the solution at a predetermined height.

また、前記細胞アッセイ用流路チップにおいて、前記培養室の容積よりも前記液溜室の容積を小さく構成し、かつ前記培養室の上部壁又は下部壁の少なくとも一方の壁は透明であることを特徴とするものである。   In the cell assay channel chip, the volume of the liquid storage chamber is configured to be smaller than the volume of the culture chamber, and at least one of the upper wall and the lower wall of the culture chamber is transparent. It is a feature.

また、前記細胞アッセイ用流路チップにおいて、前記せき止め部に、前記培養室と前記液溜室とを連通した複数の微細連通路を備えていることを特徴とするものである。   In the cell assay channel chip, the damming portion is provided with a plurality of fine communication paths communicating the culture chamber and the liquid reservoir chamber.

また、前記細胞アッセイ用流路チップにおいて、前記細胞アッセイ用流路チップ本体は、ベース本体と当該ベース本体に一体的に接合した上部プレートを備え、前記培養室及び前記液溜室は前記ベース本体に備え、前記注入口及び排出口は前記上部プレートに備えられており、前記せき止め部における堰部材（堰部）は前記ベース本体又は前記上部プレートの適宜一方或は両方に備えていることを特徴とするものである。   In the cell assay channel chip, the cell assay channel chip body includes a base body and an upper plate integrally joined to the base body, and the culture chamber and the liquid storage chamber are the base body. The inlet and the outlet are provided in the upper plate, and the dam member (dam part) in the damming part is provided in one or both of the base body and the upper plate as appropriate. It is what.

また、前記細胞アッセイ用流路チップを使用した検査装置であって、複数の細胞アッセイ用流路チップを並列して支持したチップホルダを出入自在のＣＯ２インキユベータと、当該ＣＯ２インキユベータの前側に備えられ、前記チップホルダの大きさに対応した開口部を備えると共に前記ＣＯ２インキユベータに対して出し入れされるチップホルダを支持するホルダ支持プレートと、当該ホルダ支持プレートの前記開口部に対応した位置に配置されたチップホルダに支持されている全ての細胞アッセイ用流路チップを、前記開口部を介して下側から撮像するための撮像手段と、を備えていることを特徴とするものである。 Also, the A test apparatus using cellular assays for channel chip, and CO 2 Inkiyubeta freely and out the tip holder which supports multiple simultaneous cellular assay channel chip, on the front side of the CO 2 Inkiyubeta A holder support plate that has an opening corresponding to the size of the chip holder and supports a chip holder that is inserted into and removed from the CO 2 ink evacuator; and a position corresponding to the opening of the holder support plate. And an imaging means for imaging all the cell assay flow path chips supported by the arranged chip holders from below through the openings.

本発明によれば、細胞アッセイ用流路チップにおける培養室内での溶液の流れは一方向の流れになると共に、急速な流れを抑制することができる。したがって、例えば、培養室内の細胞が舞上るようなことがなく、細胞にストレスを付与するようなことがないものである。   According to the present invention, the flow of the solution in the culture chamber in the cell assay channel chip is a one-way flow, and a rapid flow can be suppressed. Therefore, for example, the cells in the culture chamber do not fly, and the cells are not stressed.

本発明の実施形態に係る細胞アッセイ用流路チップの平面説明図である。It is a plane explanatory view of the channel chip for cell assay concerning the embodiment of the present invention. 図１におけるII−II線に沿った拡大断面説明図である。It is an expanded sectional explanatory view along the II-II line in FIG. 細胞アッセイ用流路チップの主要部を示す拡大平面説明図である。It is an expanded plane explanatory view which shows the principal part of the flow-path chip for cell assays. チップホルダに複数の細胞アッセイ用流路チップを収容した状態の説明図である。It is explanatory drawing of the state which accommodated the several flow-path chip for cell assays in the chip holder. 検査装置の構成を概念的、概略的に示した斜視説明図である。It is perspective explanatory drawing which showed the structure of the test | inspection apparatus notionally and schematically. 検査装置の構成を示す平面説明図である。It is a plane explanatory view showing the composition of an inspection device.

以下、図面を用いて本発明の実施形態に係る細胞アッセイ用流路チップについて説明するに、細胞アッセイ用流路チップ１は、図１に概念的、概略的に示すように、平面視したときに矩形状を呈する細胞アッセイ用流路チップ本体３を備えている。そして、この細胞アッセイ用流路チップ本体３の上面には複数の注入口５が備えられていると共に、各注入口５と対をなす複数の排出口７が備えられている。さらに、前記細胞アッセイ用流路チップ本体３には、前記各注入口５、排出口７のＸ，Ｙ軸方向の基準位置となる矩形状の切欠部９が適宜位置の角部に備えられている。   Hereinafter, a cell assay channel chip according to an embodiment of the present invention will be described with reference to the drawings. The cell assay channel chip 1 is viewed in plan view as schematically and schematically shown in FIG. Is provided with a cell assay flow path chip body 3 having a rectangular shape. The top surface of the cell assay flow path chip body 3 is provided with a plurality of injection ports 5 and a plurality of discharge ports 7 which are paired with the respective injection ports 5. Further, the cell assay flow path chip body 3 is provided with a rectangular notch 9 serving as a reference position in the X and Y axis directions of each of the inlets 5 and outlets 7 at an appropriate corner. Yes.

前記細胞アッセイ用流路チップ本体３に備えた前記注入口５と前記排出口７は、図２に示すように、流路１１を介して連通してある。より詳細には、前記細胞アッセイ用流路チップ本体３は、透明なベース本体（下部壁）１３の上面に透明な上部プレート（上部壁）１５を一体的に接合した構成であって、前記上部プレート１５に前記注入口５及び排出口７が形成してあり、かつ上部プレート１５の上面には、前記注入口５、排出口７に対応して溶液保持カップ１７がそれぞれ備えられている。   As shown in FIG. 2, the inlet 5 and the outlet 7 provided in the cell assay flow path chip body 3 communicate with each other through a flow path 11. More specifically, the cell assay channel chip body 3 has a structure in which a transparent upper plate (upper wall) 15 is integrally joined to an upper surface of a transparent base body (lower wall) 13, and The inlet 15 and the outlet 7 are formed in the plate 15, and a solution holding cup 17 is provided on the upper surface of the upper plate 15 corresponding to the inlet 5 and the outlet 7.

該ベース本体１３の素材は、光の透過性に優れた素材であれば特に限定はされないが、蛍光検出を行う場合は、例えば低自家蛍光ポリスチレン、紫外線透過ガラス、及びシクロオレフィン等の、自家蛍光が低い素材が好ましい。   The material of the base body 13 is not particularly limited as long as it is a material excellent in light transmittance. However, when fluorescence detection is performed, autofluorescence such as low autofluorescence polystyrene, ultraviolet transmissive glass, and cycloolefin is used. Is preferred.

前記ベース本体１３には、前記注入口５と前記排出口７とを連通した前記流路(接続路)１１が形成してあり、この接続路（流路）１１の途中には、前記注入口５から注入された溶液の流れは許容するが、溶液中の細胞１９をせき止めるためのせき止め部２１を構成する堰部２３が備えられている。すなわち、前記流路１１中に前記せき止め部２１を備えたことにより、前記流路１１は、前記注入口５に連通した上流側の培養室２５と前記排出口７に連通した下流側の液溜室２７とに区画してある。本実施例では、多くの細胞が培養可能なように、前記培養室２５の容積は前記液溜室２７の容積より大きく構成してある。   The base body 13 is formed with the flow path (connection path) 11 that connects the injection port 5 and the discharge port 7, and in the middle of the connection path (flow path) 11, the injection port Although the flow of the solution injected from 5 is allowed, a weir portion 23 constituting a damming portion 21 for damming the cells 19 in the solution is provided. That is, by providing the damming portion 21 in the channel 11, the channel 11 is connected to the upstream culture chamber 25 communicating with the inlet 5 and the downstream liquid reservoir communicating with the outlet 7. It is partitioned into a chamber 27. In this embodiment, the volume of the culture chamber 25 is configured to be larger than the volume of the liquid reservoir chamber 27 so that many cells can be cultured.

前記せき止め部２１における前記堰部２３の上面には、前記培養室２５と前記液溜室２７とを連通した複数の微細連通路２９を備えている。この微細連通路２９は、溶液の流れは許容するが、溶液内の前記細胞１９をせき止める作用をなすものであって、例えば深さは２０μｍ以下であり、幅は２００μｍ以下に形成してある。したがって、前記注入口５から細胞１９を含む溶液を注入すると、溶液中の細胞１９がせき止められて、前記培養室２５内に定着又は浮遊した状態となるものである。この際、前記培養室２５及び液溜室２７内の溶液の液面３１は前記せき止め部２１よりも常に高く保持されるものである。   A plurality of fine communication passages 29 that communicate the culture chamber 25 and the liquid storage chamber 27 are provided on the top surface of the dam portion 23 in the damming portion 21. The fine communication path 29 allows the flow of the solution, but acts to block the cells 19 in the solution. For example, the depth is 20 μm or less and the width is 200 μm or less. Therefore, when a solution containing cells 19 is injected from the injection port 5, the cells 19 in the solution are blocked and become fixed or floating in the culture chamber 25. At this time, the liquid level 31 of the solution in the culture chamber 25 and the liquid storage chamber 27 is always kept higher than the damming portion 21.

換言すれば、前記せき止め部２１は常に溶液内に没入した状態に保持されるものであって、前記微細連通路２９の流出側（前記液溜室２７側の出口）が溶液の液面３１よりも高くなるようなことがなく、微細連通路２９の出口側には前記溶液溜室２７の液面３１が高いことに起因する液圧が、常に背圧として作用しているものである。よって、前記液溜室２７は一種の背圧付与室を構成するものである。   In other words, the damming portion 21 is always kept immersed in the solution, and the outflow side of the fine communication passage 29 (the outlet on the liquid reservoir 27 side) is more than the liquid level 31 of the solution. The liquid pressure resulting from the high liquid surface 31 of the solution reservoir 27 is always acting as a back pressure on the outlet side of the fine communication passage 29. Therefore, the liquid reservoir 27 constitutes a kind of back pressure applying chamber.

前記構成により、前記培養室２５に対する培地や試薬溶液などの溶液の注入は注入口５側から行われ、溶液の排出は排出口７側で行われるものである。細胞培養用の培地は、細胞アッセイに使用する細胞の生育に適した液体培地である限り、特に限定はされない。したがって、溶液の交換を行うときは、前記注入口５から新しい溶液を注入し、排出口７側において排出することにより容易に行い得るものである。この際、溶液の流れは注入口５側から排出口７側への一定方向の流れであり、かつ前記培養室２５及び液溜室２７の液面３１を前記せき止め部２１よりも高く保持して溶液の交換を行うことが可能である。   With the configuration described above, the injection of a solution such as a medium or a reagent solution into the culture chamber 25 is performed from the injection port 5 side, and the solution is discharged from the discharge port 7 side. The medium for cell culture is not particularly limited as long as it is a liquid medium suitable for the growth of cells used in the cell assay. Therefore, when exchanging the solution, it can be easily performed by injecting a new solution from the injection port 5 and discharging it at the discharge port 7 side. At this time, the flow of the solution is a flow in a certain direction from the inlet 5 side to the outlet 7 side, and the liquid surface 31 of the culture chamber 25 and the liquid reservoir chamber 27 is held higher than the damming portion 21. It is possible to exchange the solution.

よって、前記せき止め部２１における前記微細連通路２９の出口側には、前記液溜室２７内の液圧が背圧として作用するので、前記微細連通路２９における溶液の流速が低速に保持されるものである。したがって、例えば、前記培養室２５内における溶液の交換による乱流等の発生が抑制されるため、前記培養室２５内で培養されている細胞１９に物理的なストレス等を付与することなく、溶液の交換を行い得るものである。   Therefore, since the liquid pressure in the liquid reservoir 27 acts as a back pressure on the outlet side of the fine communication passage 29 in the damming portion 21, the flow rate of the solution in the fine communication passage 29 is maintained at a low speed. Is. Therefore, for example, since the occurrence of turbulence or the like due to the exchange of the solution in the culture chamber 25 is suppressed, the solution without applying physical stress or the like to the cells 19 cultured in the culture chamber 25 Can be exchanged.

なお、前記構成においては、前記せき止め部２１の堰部２３をベース本体１３に備えた場合について例示したが、前記堰部２３は上部プレート１５に備えた構成とすることも可能である。この場合、せき止め部２１における前記微細連通路２９は、堰部２３の下面に備えた構成となるものである。また、ベース本体１３及び上部プレート１５にそれぞれ中間高さの堰部２３を備え、この上下の堰部２３の間に微細通路２９を形成することも可能である。   In addition, in the said structure, although illustrated about the case where the base body 13 was equipped with the dam part 23 of the said damming part 21, the said dam part 23 can also be set as the structure with which the upper plate 15 was equipped. In this case, the fine communication passage 29 in the damming portion 21 is provided on the lower surface of the dam portion 23. It is also possible to provide the base body 13 and the upper plate 15 with dam portions 23 having intermediate heights, and to form a fine passage 29 between the upper and lower dam portions 23.

前記細胞アッセイ用流路チップ１をＣＯ２インキユベータ内に収納して細胞１９の培養を行うために、細胞アッセイ用流路チップ１を保持するチップホルダ３３が備えられている。チップホルダ３３は、図４に示すように、複数の細胞アッセイ用流路チップ１を並列した状態に収容保持するための複数の収容部３５を備えている。上記各収容部３５は、収容した細胞アッセイ用流路チップ１を下側から観察可能に構成してあり、かつ各収容部３５には、細胞アッセイ用流路チップ１における前記切欠部９と係合して位置決めを行う位置決め突出部３７が備えられている。 A chip holder 33 for holding the cell assay flow path chip 1 is provided to house the cell assay flow path chip 1 in a CO 2 incubator and culture the cells 19. As shown in FIG. 4, the chip holder 33 includes a plurality of accommodating portions 35 for accommodating and holding a plurality of cell assay flow path chips 1 in parallel. Each of the accommodating portions 35 is configured so that the accommodated cell assay channel chip 1 can be observed from below, and each of the accommodating units 35 is associated with the notch 9 in the cell assay channel chip 1. A positioning projection 37 for positioning is provided.

したがって、前記チップホルダ３３における各収容部３５に細胞アッセイ用流路チップ１をそれぞれ収容すると、複数の細胞アッセイ用流路チップ１は、前記位置決め突出部３７に切欠部９を係合して位置決めされることになる。よって、各細胞アッセイ用流路チップ１の方向性は全て同一方向性に揃えられた状態にあるものである。   Therefore, when the cell assay channel chip 1 is accommodated in each accommodating part 35 in the chip holder 33, the plurality of cell assay channel chips 1 are positioned by engaging the notch portions 9 with the positioning protrusions 37. Will be. Therefore, the directionality of each cell assay channel chip 1 is in a state where all the directionality is aligned.

前記細胞アッセイ用流路チップ１を用いて培養した細胞１９の検査を行うための検査装置は、次のごとき構成である。すなわち、図５、図６に概念的、概略的に示すように、検査装置３９は箱状に構成してあって、前記チップホルダ３３を出入自在のＣＯ２インキユベータ４１を備えている。そして、前記ＣＯ２インキユベータ４１の前側には、前記チップホルダ３３を支持するホルダ支持プレート４３が備えられている。このホルダ支持プレート４３には、前記チップホルダ３３の大きさに対応した開口部４５が備えられている。そして、前記ホルダ支持プレート４３の下方位置には、前記開口部４５に対応して位置決めしたチップホルダ３３に支持されている全ての細胞アッセイ用流路チップ１の照明を行うための適数の光源４７が備えられていると共に、前記開口部４５を介して全ての細胞アッセイ用流路チップ１を下側から撮像するＣＣＤカメラなどの撮像手段４９が備えられている。 An inspection apparatus for inspecting cells 19 cultured using the cell assay flow path chip 1 has the following configuration. That is, as conceptually and schematically shown in FIG. 5 and FIG. 6, the inspection device 39 is configured in a box shape and includes a CO 2 incubator 41 through which the chip holder 33 can be moved in and out. A holder support plate 43 that supports the chip holder 33 is provided on the front side of the CO 2 ink elevator 41. The holder support plate 43 is provided with an opening 45 corresponding to the size of the chip holder 33. An appropriate number of light sources for illuminating all the cell assay flow path chips 1 supported by the chip holder 33 positioned corresponding to the opening 45 are located below the holder support plate 43. 47 and an imaging means 49 such as a CCD camera for imaging all the cell assay channel chips 1 from the lower side through the opening 45.

したがって、前記ＣＯ２インキユベータ４１において培養した後に、チップホルダ３３に保持されている全てのチップホルダ３３を撮像手段４９によって撮像し、細胞の生理活性の観察のための光学的検出を行うことができるものである。なお、撮像手段４９によって細胞アッセイ用流路チップ１の撮像を行う際には、全ての細胞アッセイ用流路チップ１を同時に撮像することが望ましい。しかし、個別に又は連続的に撮像することも可能である。 Therefore, after culturing in the CO 2 incubator 41, all the chip holders 33 held by the chip holder 33 can be imaged by the imaging means 49, and optical detection for observing the physiological activity of the cells can be performed. Is. When imaging the cell assay channel chip 1 by the imaging means 49, it is desirable to image all the cell assay channel chips 1 simultaneously. However, it is also possible to image individually or continuously.

該光学的検出の例としては、蛍光検出等の発光検出、吸光度検出などが挙げられる。   Examples of the optical detection include luminescence detection such as fluorescence detection, and absorbance detection.

本発明において細胞は、付着性細胞等の任意の細胞でよい。細胞の具体的な例としては、癌細胞、神経細胞、肝細胞、繊維芽細胞、筋芽細胞、平滑筋細胞、心筋細胞、骨格筋細胞、幹細胞、中胚葉系幹細胞、胚性幹細胞、グリア細胞、胎性幹細胞、造血幹細胞、肥満細胞、脂肪細胞、神経幹細胞、及び血球細胞などが挙げられる。   In the present invention, the cell may be any cell such as an adherent cell. Specific examples of cells include cancer cells, nerve cells, hepatocytes, fibroblasts, myoblasts, smooth muscle cells, cardiomyocytes, skeletal muscle cells, stem cells, mesodermal stem cells, embryonic stem cells, glial cells Embryonic stem cells, hematopoietic stem cells, mast cells, adipocytes, neural stem cells, blood cells and the like.

また、細胞アッセイには、細胞の生理活性等、細胞の種々の特性の測定を包含する。細胞アッセイの具体的な例としては、エストロゲン応答アッセイ、アセチルコリンステラーゼアッセイ等の神経細胞分化アッセイ、乳酸デヒドロゲナーゼアッセイ、ＤＮＡ細分化アッセイ等のアポトーシスアッセイ、抗アレルギーアッセイ、ＨＳＰ４７細胞アッセイ等のストレス応答・回復アッセイ、神経細胞保護アッセイ等の抗アルツハイマー症アッセイ、α―グルコシダーゼ阻害アッセイ等の糖吸収抑制アッセイ、ＭＴＴアッセイ等の抗腫瘍アッセイ、ニュートラルレッドアッセイ等の細胞毒性アッセイ、ＰＣ１２細胞等を用いる神経細胞伸張アッセイ、アルカリフォスファターゼ活性化アッセイ、脂肪細胞分化アッセイ等の脂肪生成アッセイ、毛乳頭細胞成長アッセイ、及びメラニン生成抑制アッセイ等の皮膚美白アッセイなどが挙げられる。   In addition, the cell assay includes measurement of various characteristics of the cell such as the physiological activity of the cell. Specific examples of cell assays include stress response and recovery such as estrogen response assay, neuronal differentiation assay such as acetylcholinesterase assay, apoptosis assay such as lactate dehydrogenase assay, DNA fragmentation assay, anti-allergy assay, HSP47 cell assay, etc. Anti-Alzheimer's disease assay such as assay, neuronal protection assay, sugar absorption inhibition assay such as α-glucosidase inhibition assay, antitumor assay such as MTT assay, cytotoxicity assay such as neutral red assay, neuronal cell extension using PC12 cells, etc. Adipogenesis assay such as assay, alkaline phosphatase activation assay, adipocyte differentiation assay, skin whitening assay such as dermal papilla cell growth assay, and melanogenesis inhibition assay It is.

１ 細胞アッセイ用流路チップ
３ 細胞アッセイ用流路チップ本体
５ 注入口
７ 排出口
９ 切欠部
１１ 流路
１３ ベース本体
１５ 上部プレート
１７ 溶液保持カップ
１９ 細胞
２１ せき止め部
２３ 堰部
２５ 培養室
２７ 液溜室
２９ 微細連通路
３３ チップホルダ
３５ 収容部
３９ 検査装置
４３ ホルダ支持プレート
４５ 開口部
４９ 撮像手段 DESCRIPTION OF SYMBOLS 1 Cell assay flow path chip 3 Cell assay flow path chip main body 5 Inlet 7 Discharge port 9 Notch 11 Flow path 13 Base main body 15 Upper plate 17 Solution holding cup 19 Cell 21 Damping part 23 Weir part 25 Culture chamber 27 Liquid Reservoir chamber 29 Fine communication path 33 Chip holder 35 Storage section 39 Inspection device 43 Holder support plate 45 Opening section 49 Imaging means

Claims (6)

細胞培養が可能な細胞アッセイ用流路チップであって、当該細胞アッセイ用流路チップ本体の上面に溶液の注入口と排出口とを備え、前記注入口と排出口とを連通した流路中に、前記注入口から注入された溶液内の細胞をせき止めするためのせき止め部を備え、当該せき止め部の上流側に培養室を備えると共に、前記せき止め部の下流側に、前記せき止め部を流れる溶液の流速を低速に抑制するために、細胞培養時には前記せき止め部よりも液面高さを高く保持する液溜室を備えていることを特徴とする細胞アッセイ用流路チップ。   A cell assay channel chip capable of culturing cells, wherein the cell assay channel chip has a solution inlet and an outlet on the upper surface of the channel chip body, and the channel and the outlet are in communication with each other. And a damming portion for damaging cells in the solution injected from the inlet, a culture chamber provided upstream of the damming portion, and flowing through the damming portion downstream of the damming portion. In order to suppress the flow rate of the cell at low speed, the cell assay channel chip is provided with a liquid reservoir chamber that holds a liquid surface height higher than that of the damming portion during cell culture. 請求項１に記載の細胞アッセイ用流路チップにおいて、前記注入口及び排出口に、溶液を所定高さに保持する溶液保持カップを備えていることを特徴とする細胞アッセイ用流路チップ。   2. The cell assay channel chip according to claim 1, further comprising a solution holding cup that holds the solution at a predetermined height at the inlet and the outlet. 3. 請求項１又は２に記載の細胞アッセイ用流路チップにおいて、前記培養室の容積よりも前記液溜室の容積を小さく構成し、かつ前記培養室の上部壁又は下部壁の少なくとも一方の壁は透明であることを特徴とする細胞アッセイ用流路チップ。   The flow path chip for cell assay according to claim 1 or 2, wherein the volume of the liquid storage chamber is smaller than the volume of the culture chamber, and at least one wall of the upper wall or the lower wall of the culture chamber is A channel chip for cell assay, which is transparent. 請求項１、２又は３に記載の細胞アッセイ用流路チップにおいて、前記せき止め部に、前記培養室と前記液溜室とを連通した複数の微細連通路を備えていることを特徴とする細胞アッセイ用流路チップ。   4. The cell assay channel chip according to claim 1, 2 or 3, wherein the cell is provided with a plurality of fine communication passages that communicate the culture chamber and the liquid storage chamber in the damming portion. Flow path chip for assay. 請求項１〜４のいずれかに記載の細胞アッセイ用流路チップにおいて、前記細胞アッセイ用流路チップ本体は、ベース本体と当該ベース本体に一体的に接合した上部プレートを備え、前記培養室及び前記液溜室は前記ベース本体に備え、前記注入口及び排出口は前記上部プレートに備えられており、前記せき止め部における堰部材は前記ベース本体又は前記上部プレートの適宜一方或は両方に備えていることを特徴とする細胞アッセイ用流路チップ。   5. The cell assay channel chip according to claim 1, wherein the cell assay channel chip body includes a base body and an upper plate integrally joined to the base body, and the culture chamber and The liquid reservoir is provided in the base body, the inlet and the outlet are provided in the upper plate, and the dam member in the damming portion is provided in one or both of the base body and the upper plate as appropriate. A channel chip for cell assay, characterized in that 請求項１〜５のいずれかに記載の細胞アッセイ用流路チップを使用した検査装置であって、複数の細胞アッセイ用流路チップを並列して支持したチップホルダを出入自在のＣＯ２インキユベータと、当該ＣＯ２インキユベータの前側に備えられ、前記チップホルダの大きさに対応した開口部を備えると共に前記ＣＯ２インキユベータに対して出し入れされるチップホルダを支持するホルダ支持プレートと、当該ホルダ支持プレートの前記開口部に対応した位置に配置されたチップホルダに支持されている全ての細胞アッセイ用流路チップを、前記開口部を介して下側から撮像するための撮像手段と、を備えていることを特徴とする検査装置。 A test apparatus using the cell assay flow path chip according to any one of claims 1 to 5, wherein a CO 2 ink incubator is provided with a chip holder that supports a plurality of cell assay flow path chips in parallel. A holder support plate which is provided on the front side of the CO 2 ink evacuator and has an opening corresponding to the size of the chip holder and which supports a chip holder which is taken in and out of the CO 2 ink evacuator, and the holder support plate Imaging means for imaging all the cell assay flow path chips supported by the chip holder disposed at a position corresponding to the opening from below through the opening. Inspection device characterized by
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