JP3455262B2 - Optical immunoassay and device - Google Patents
Optical immunoassay and deviceInfo
- Publication number
- JP3455262B2 JP3455262B2 JP32852893A JP32852893A JP3455262B2 JP 3455262 B2 JP3455262 B2 JP 3455262B2 JP 32852893 A JP32852893 A JP 32852893A JP 32852893 A JP32852893 A JP 32852893A JP 3455262 B2 JP3455262 B2 JP 3455262B2
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- Prior art keywords
- light
- antigen
- thin film
- antibody
- layer
- Prior art date
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- Investigating Or Analysing Materials By Optical Means (AREA)
Description
【０００１】[0001]
【産業上の利用分野】本発明は、光学的免疫測定法およ
びそのデバイスに関する。FIELD OF THE INVENTION The present invention relates to an optical immunoassay and its device.
【０００２】[0002]
【従来の技術】生体関連物質の定量法として、放射性物
質や酵素で標識した抗原もしくは抗体を用いたラジオイ
ムノアッセイ法や酵素免疫測定法は、その反応系に抗原
抗体反応を用いることから特定のタンパク質や生理活性
物質の測定が可能となり、既に一部は病理診断技術とし
て確立している。2. Description of the Related Art Radioimmunoassays and enzyme immunoassays, which use an antigen or antibody labeled with a radioactive substance or an enzyme as a method for quantifying a biological substance, use an antigen-antibody reaction in the reaction system, and And physiologically active substances can be measured, and some of them have already been established as pathological diagnosis techniques.
【０００３】しかし、これらの従来技術は、抗原もしく
は抗体に結合した放射性物質や酵素などの標識物質の量
を測定し、その結果から抗原もしくは抗体の濃度を定量
するものであった。そのため、単に抗原抗体反応だけで
なく、標識物質を抗原もしくは抗体に結合させるための
操作が必要となり、同時に標識物質を測定するための高
価な設備も要求された。However, these conventional techniques measure the amount of a labeling substance such as a radioactive substance or an enzyme bound to an antigen or an antibody, and quantify the concentration of the antigen or the antibody from the result. Therefore, not only the antigen-antibody reaction but also an operation for binding the labeling substance to the antigen or the antibody is required, and at the same time, expensive equipment for measuring the labeling substance is required.
【０００４】さらに、臨床検査の主流ともいえるラジオ
イムノアッセイ法では、抗原もしくは抗体の標識に放射
性物質を用いることから、その操作は専門の技術者に委
ねられ、また放射性物質の保管や廃棄には厳重な注意を
必要とされている。Furthermore, in the radioimmunoassay method, which can be said to be the mainstream of clinical tests, a radioactive substance is used for labeling an antigen or an antibody, so that the operation is entrusted to a professional engineer, and the storage and disposal of the radioactive substance are strictly performed. Attention is needed.
【０００５】これらの問題を解決するため、光干渉性基
板の表面を抗原抗体反応の場とし、そこで生じる有機物
質の厚みの増加を色調の変化として検出する、または楕
円偏光測定器を用いて定量する光学的免疫測定法がいく
つか提案されている。In order to solve these problems, the surface of the light-interfering substrate is used as a field for an antigen-antibody reaction, and an increase in the thickness of the organic substance generated there is detected as a change in color tone, or is quantified using an ellipsometer. Several optical immunoassays have been proposed.
【０００６】例えば、特公平４−８７４２号公報には、
光反射性基板の上に光干渉層を積層し、さらに反応性中
間層を設けて抗原または抗体を固定化したデバイスが開
示されており、この表面で抗原抗体反応を生じせしめ、
その結果生じる有機物質の厚みの増加に伴う色調変化を
検出することにより、結合した抗原または抗体の有無を
判断するものである。ここでいう厚みの増加とは、抗原
抗体反応により結合してなる実質的に単分子層の抗原ま
たは抗体の大きさであり、そのため色調の変化は単分子
層相当の厚みの有無にのみ起因することから、その定量
には至っていない。[0006] For example, Japanese Patent Publication No. 4-8742 discloses that
A device in which an optical interference layer is laminated on a light-reflecting substrate, and a reactive intermediate layer is further provided to immobilize an antigen or an antibody is disclosed, and an antigen-antibody reaction is caused on this surface,
The presence or absence of the bound antigen or antibody is determined by detecting the resulting change in color tone with an increase in the thickness of the organic substance. The increase in thickness here is the size of an antigen or antibody in a substantially monolayer formed by binding by an antigen-antibody reaction, and therefore the change in color tone is caused only by the presence or absence of a thickness equivalent to a monolayer. Therefore, its quantification has not been reached.
【０００７】また、ここでいう光干渉層は、ＬＢ法を用
いてステアリン酸系有機物質の単分子層を数十層積層し
たものであり、その厚みで光学的特性が決定されること
から、その表面に設けられる前記反応層の厚みも厳しく
制限される。さらに、ＬＢ法を用いた製膜では大きな面
積の基板に対応できずまた量産性もなく、多層を積層し
たときには崩れ易いなどの問題を有しており、本来の目
的である簡便な測定法とそのデバイスとを広く提供する
には至っていない。なお、光干渉層として種々の無機材
料も示されているが、その構成に係る具体例は何等開示
されていない。Further, the optical interference layer mentioned here is formed by laminating several tens of monomolecular layers of a stearic acid type organic substance by using the LB method, and since the optical characteristics are determined by the thickness, The thickness of the reaction layer provided on the surface is also severely limited. Furthermore, the film formation using the LB method has a problem that it cannot be applied to a substrate having a large area, mass productivity is low, and it easily breaks when multiple layers are laminated. It has not been widely offered with the device. Although various inorganic materials are shown as the light interference layer, no specific example relating to the structure is disclosed.
【０００８】一方、光干渉層に無機材料を用いた技術
は、ジョン・ハンリン（Ｊｏｈｎ Ｈａｎｌｉｎ）著
“薄膜（ＴＨＩＮ ＦＩＬＭＳ）”と題する論文：フォ
トニクス・スペクトラ（ＰＨＯＴＯＮＩＣＳ・ＳＰＥＣ
ＴＲＡ）誌（Ｆebruary, １９８８）やＰＣＴ／ＵＳ９
０／０５３１６号にも開示されている。これらの技術
は、シリコンウェハーの表面に一酸化シリコンと二酸化
シリコンとを積層した後、前記した特公平４−８７４２
号公報と同様に抗原または抗体を固定化するための反応
層を設けるものであり、楕円偏光測定器を用いた場合に
は定量方法も開示されているが、目視による色調変化か
らは抗原抗体反応による結合物質の有無しか判定できて
いない。On the other hand, a technique using an inorganic material for the optical interference layer is described in a paper entitled "THIN FILMS" by John Hanlin: Photonics SPEC.
TRA) magazine (February, 1988) and PCT / US9
It is also disclosed in 0/05316. According to these techniques, after laminating silicon monoxide and silicon dioxide on the surface of a silicon wafer, the above-mentioned Japanese Patent Publication No. 4-8742 is used.
Similar to the publication, it provides a reaction layer for immobilizing an antigen or antibody, and a quantification method is also disclosed when an ellipsometer is used. Only the presence or absence of the binding substance can be determined by.
【０００９】[0009]
【発明が解決しようとする課題】そこで、本発明は放射
性物質などを必要としないだけでなく、抗原抗体判定以
外の操作を伴わず、抗原抗体反応により結合した有機物
質の量の大小を目視による色調変化から判断できる簡便
な光学的免疫測定法を提供するものであり、そのデバイ
スを安価でかつ大量に提供することを目的としている。Therefore, the present invention not only requires no radioactive substance, but also visually confirms the amount of the organic substance bound by the antigen-antibody reaction without any operation other than the antigen-antibody determination. It is intended to provide a simple optical immunoassay that can be judged from a change in color tone, and an object thereof is to provide a large amount of the device at a low cost.
【００１０】[0010]
【課題を解決するための手段】すなわち、本発明による
光学的免疫測定法は、光反射層上に、屈折率がその表面
に近づくほど小さくなる多層構造を有すると共に、可視
光を吸収する光干渉層を形成して、その表面に抗原又は
抗体のいずれか一方である第１の有機物質を予め単分子
層を形成する分子数以下の量を吸着させておき、吸着さ
せた上記第１の有機物質と抗原抗体反応を生ずる抗体又
は抗原のいずれかである第２の有機物質を与えて抗原抗
体反応を生じさせ、その抗原抗体反応の結果生ずる光干
渉層表面での有機物質密度の増加を可視光領域での光干
渉色の変化として定量的に検出するものである。That is, in the optical immunoassay method according to the present invention, the refractive index is provided on the surface of the light reflecting layer.
Has a multi-layered structure that becomes smaller as it approaches, and a light interference layer that absorbs visible light is formed, and a first organic substance, which is either an antigen or an antibody, is previously formed as a single molecule on its surface.
An antibody or an antibody that causes an antigen-antibody reaction with the adsorbed first organic substance is adsorbed in an amount equal to or less than the number of molecules forming a layer.
The cause an antigen-antibody reaction gives a second organic material is either antigen, the antigen-antibody reaction occurring upon an increase in organic matter density of the optical interference layer surface of the optical interference color in the visible light region The change is quantitatively detected.
【００１１】また、そのためのデバイスは、光反射性固
体表面に、屈折率がその表面に近づくほど小さくなる多
層構造を有すると共に、可視光が吸収されかつ有機物質
が吸着される表面を有する光干渉層を設け、光反射性固
体表面は光干渉層表面よりも大きな屈折率を有する材料
で構成し、光干渉層の表面に単分子層を形成する分子数
以下の量の抗原もしくは抗体を吸着させたことを特徴と
する。Further, a device therefor often has a light-reflecting solid surface whose refractive index becomes smaller as it gets closer to the surface.
In addition to having a layer structure, a light interference layer having a surface that absorbs visible light and adsorbs an organic substance is provided, and the light-reflecting solid surface is composed of a material having a refractive index larger than that of the light interference layer surface. Number of molecules forming a monolayer on the surface of the interference layer
It is characterized in that the following amount of antigen or antibody is adsorbed.
【００１２】さらに、光干渉層を、プラズマ反応法で形
成することも特徴としている。Further, the optical interference layer is characterized by being formed by a plasma reaction method.
【００１３】[0013]
【作用】本発明による以上の構成は、デバイス表面での
抗原抗体反応による有機物質のわずかな密度増加を光学
的膜厚の増加として捉え、それを目視による色調変化と
して定量的に検出することを可能とするものであり、同
時にそのデバイスを安価で大量に供給することも可能に
するものである。The above configuration according to the present invention is to detect a slight increase in the density of the organic substance due to the antigen-antibody reaction on the device surface as an increase in the optical film thickness, and to detect it quantitatively as a visual color change. It is also possible to supply the device at a low cost and in large quantities at the same time.
【００１４】すなわち、光干渉層の下層から表面に至る
屈折率を順次に小さくなる多層構造にしたことから反射
分光特性に優れるだけでなく、その下地になる光反射性
固体表面の屈折率を光干渉層表面の屈折率よりも大きく
したため、色調変化が鮮明になる。That is, since the refractive index from the lower layer to the surface of the light interference layer is made smaller in order, the reflection spectral characteristics are excellent, and the refractive index of the surface of the light-reflective solid layer underlying the light-reflecting layer is Since the refractive index of the interference layer is made larger than that of the surface of the interference layer, the change in color tone becomes clear.
【００１５】さらに、本発明の構成は光干渉層の表面に
有機物質としての抗原を低密度に吸着させることから、
従来技術による単分子層としての膜厚増加の有無ではな
く、色調変化に基づく定量的な検出が可能である。Further, in the constitution of the present invention, since the antigen as the organic substance is adsorbed at a low density on the surface of the light interference layer,
Quantitative detection based on a change in color tone is possible, regardless of whether or not the thickness of the monomolecular layer is increased by the conventional technique.
【００１６】同時に、従来技術で必要とした反応性中間
層が不要となり、また従来はＬＢ法を用いていた光干渉
層を量産技術として実績を持つプラズマ反応法で連続的
に形成できることから、大型基板を用いた大量生産も可
能とする。At the same time, the reactive intermediate layer required in the prior art is no longer necessary, and the optical interference layer, which has conventionally used the LB method, can be continuously formed by the plasma reaction method, which has a proven track record as a mass production technology. Mass production using substrates is also possible.
【００１７】[0017]
【実施例】本発明を、図面および実験データに基づき詳
細に説明する。The present invention will be described in detail with reference to the drawings and experimental data.
【００１８】光干渉層の表面に、抗原または抗体が単分
子層の厚みで付着したとき、その色調がどのように変化
するのかを簡便に調べるため、プラズマ反応法を用いて
製膜した光干渉層の多層構造表面に、アクリロニトリル
（以下、ＡＮとする）薄膜を同じくプラズマ反応法によ
り堆積させ、その堆積前後の目視による色調、反射分光
特性および色度（以下、これらを光学特性とする）を調
べた。なお、有機物質としてＡＮ薄膜を選定したのは、
それが抗原や抗体など一般のタンパクと同じ屈折率を有
するためであり、その膜厚も抗体の単分子層を想定して
約８nmとした。In order to easily examine how the color tone changes when an antigen or antibody is attached to the surface of the light interference layer in the thickness of a monolayer, the light interference formed by a plasma reaction method is used. An acrylonitrile (hereinafter referred to as AN) thin film is similarly deposited on the surface of the multi-layer structure by the plasma reaction method, and the color tone, reflection spectral characteristic and chromaticity (hereinafter, these optical characteristics) are visually observed before and after the deposition. Examined. The AN thin film was selected as the organic substance.
This is because it has the same refractive index as general proteins such as antigens and antibodies, and its film thickness was set to about 8 nm assuming a monolayer of the antibody.
【００１９】まず、光反射性固体表面の構成と光学特性
について調べた。ここで、光反射性固体として、基板に
はスライドガラスを用い、その表面に電子ビーム蒸着法
を用いて厚さが２５０nmのクロム薄膜を堆積させて光反
射性固体表面とした。なお、比較のため、クロム薄膜を
設けないスライドガラスおよびシリコンウェハーについ
ても同様に調べた。これらの基板を図１に示す製膜チャ
ンバーに設置し、チャンバー内を５×１０-3Ｐaの圧力
まで排気した後、モノシラン、アルゴンおよび亜酸化窒
素などの反応ガスを３３Ｐaの圧力まで導入し、プラズ
マ反応法によりシリコン、酸化シリコンおよび二酸化シ
リコンの３層を連続的に堆積させ、この多層構造を光干
渉層とした。図２の（ａ）には、この多層構造の断面図
を示す。なお、図２の（ｂ）には、クロム薄膜１２を設
けない例（比較例１）を、また図２の（ｃ）にはシリコ
ンウェハー１３’を用いた例（比較例２）を示してい
る。First, the structure and optical characteristics of the light-reflecting solid surface were investigated. Here, as the light-reflective solid, a glass slide was used as a substrate, and a chromium thin film having a thickness of 250 nm was deposited on the surface thereof by an electron beam evaporation method to obtain a light-reflective solid surface. For comparison, slide glasses and silicon wafers not provided with a chromium thin film were also examined in the same manner. These substrates were placed in the film forming chamber shown in FIG. 1, the chamber was evacuated to a pressure of 5 × 10 −3 Pa, and then reaction gases such as monosilane, argon and nitrous oxide were introduced up to a pressure of 33 Pa, Three layers of silicon, silicon oxide and silicon dioxide were successively deposited by the plasma reaction method, and this multilayer structure was used as an optical interference layer. FIG. 2A shows a sectional view of this multilayer structure. 2B shows an example in which the chromium thin film 12 is not provided (Comparative Example 1), and FIG. 2C shows an example in which the silicon wafer 13 'is used (Comparative Example 2). There is.
【００２０】これら薄膜の組成は、モノシランと亜酸化
窒素との導入割合を変えることにより制御した。また、
各薄膜の膜厚は製膜時間で制御し、シリコン薄膜１３は
５６０nm、酸化シリコン薄膜１４は１３０nm、また二酸
化シリコン薄膜１５は２０nmとした。なお、各薄膜の屈
折率を楕円偏光測定器で調べた結果、シリコンは４．
０、酸化シリコンは２．０、また二酸化シリコンは１．
５であった。The composition of these thin films was controlled by changing the introduction ratio of monosilane and nitrous oxide. Also,
The film thickness of each thin film was controlled by the film formation time, and the silicon thin film 13 was 560 nm, the silicon oxide thin film 14 was 130 nm, and the silicon dioxide thin film 15 was 20 nm. In addition, as a result of examining the refractive index of each thin film with an ellipsometer, it was found that silicon was 4.
0, silicon oxide is 2.0, and silicon dioxide is 1.
It was 5.
【００２１】これら試料の光学的特性を調べた後、前記
と同じ製膜チャンバーに試料を設置し、３７Ｐaの圧力
までＡＮモノマーを導入してプラズマ反応法によりＡＮ
薄膜を堆積させ、同じく光学的特性を調べた。その結果
は、表１に示すように、光反射性固体表面にクロム薄膜
を設けたとき、ＡＮ薄膜堆積前後での色調変化が最も大
きかった。このクロム薄膜の有無による光学特性の違い
は、光干渉層の下地に光反射性表面が必要なことを示し
ている。また、クロム薄膜に対するシリコンウェハーで
の感度が低いのは、シリコンウェハー自体の色調が干渉
色に現れるためである。After investigating the optical characteristics of these samples, the samples were placed in the same film forming chamber as described above, and AN monomer was introduced to a pressure of 37 Pa to carry out AN reaction by plasma reaction method.
Thin films were deposited and also examined for optical properties. As a result, as shown in Table 1, when the chromium thin film was provided on the light-reflecting solid surface, the largest change in color tone was observed before and after the AN thin film was deposited. The difference in the optical characteristics depending on the presence or absence of the chromium thin film indicates that a light-reflecting surface is required as the base of the light interference layer. Further, the reason why the sensitivity of the silicon wafer to the chromium thin film is low is that the color tone of the silicon wafer itself appears in the interference color.
【００２２】[0022]
【表１】 ＡＮ薄膜着膜前 ＡＮ薄膜着膜後 光反射性固体表面 色調 Ｘ座標 Ｙ座標 吸収波長 色調 Ｘ座標 Ｙ座標 吸収波長 クロム薄膜 黄 0.44 0.45 456 桃 0.47 0.42 476 （本発明） スライドガラス 黄 0.43 0.46 450nm 橙 0.44 0.46 459nm （比較例１） シリコンウェハー 赤黄 0.43 0.45 455 赤橙 0.46 0.52 473 （比較例２）[Table 1] Before AN thin film deposition After AN thin film deposition Light-reflecting solid surface Color tone X coordinate Y coordinate Absorption wavelength Color tone X coordinate Y coordinate Absorption wavelength Chrome thin film Yellow 0.44 0.45 456 Pink 0.47 0.42 476 (Invention) Glass slide Yellow 0.43 0.46 450nm Orange 0.44 0.46 459nm (Comparative Example 1) Silicon wafer Red yellow 0.43 0.45 455 Red orange 0.46 0.52 473 (Comparative example 2)
【００２３】光反射性固体表面を構成する金属薄膜材料
と光学特性との関係を調べた。用いた金属は、クロム、
マンガン、チタン、ニッケル、銅およびアルミであり、
その製膜方法や膜厚は前記のクロムのときと同じであ
る。なお、これら薄膜の屈折率を楕円偏光測定器で調べ
たところ、クロムは２．６、マンガンとチタンは２．
０、ニッケルは１．５、銅は０．７７、そしてアルミは
０．６６であった。The relationship between the metal thin film material constituting the light-reflecting solid surface and the optical characteristics was investigated. The metal used is chromium,
Manganese, titanium, nickel, copper and aluminum,
The film forming method and the film thickness are the same as in the case of chromium. When the refractive index of these thin films was examined by an ellipsometer, chromium was 2.6 and manganese and titanium were 2.
0, nickel 1.5, copper 0.77, and aluminum 0.66.
【００２４】これらの金属薄膜表面を光反射性固体表面
として、前記同様に光干渉層およびＡＮ薄膜を設け、そ
の光学特性を調べた。このとき、光干渉層の多層構造
は、下層からシリコンが２４nm、酸化シリコンが４２n
m、また二酸化シリコンが２０nmとした。Optical interference layers and an AN thin film were provided in the same manner as above using these metal thin film surfaces as light reflecting solid surfaces, and the optical characteristics thereof were examined. At this time, the multilayer structure of the optical interference layer is such that the silicon is 24 nm and the silicon oxide is 42 n from the lower layer.
m, and silicon dioxide was 20 nm.
【００２５】得られた結果は表２のごとくであり、クロ
ム、マンガンおよびチタンを光反射性固体表面としたと
きには鮮明な色調変化が認められるが、ニッケル、銅お
よびアルミでは色調変化が小さかった。これは、光反射
性固体表面の屈折率と光干渉層の光学的特性との相互関
係によると考えられ、光干渉層の表面を構成する二酸化
シリコンの屈折率が１．５であったことを考慮すると、
光反射性固体表面の屈折率は、光干渉層表面の屈折率よ
りも大きくする必要がある。The results obtained are as shown in Table 2, and when chromium, manganese, and titanium were used as the light-reflecting solid surface, a clear change in color tone was observed, but with nickel, copper, and aluminum, the change in color tone was small. This is considered to be due to the interrelationship between the refractive index of the light-reflecting solid surface and the optical characteristics of the light interference layer, and it was confirmed that the refractive index of silicon dioxide forming the surface of the light interference layer was 1.5. Considering
The refractive index of the light-reflecting solid surface needs to be higher than that of the light interference layer surface.
【００２６】[0026]
【表２】 ＡＮ薄膜着膜前 ＡＮ薄膜着膜後 光反射性固体表面 色調 Ｘ座標 Ｙ座標 吸収波長 色調 Ｘ座標 Ｙ座標 吸収波長 クロム薄膜 黄 0.39 0.45 452nm 赤紫 0.41 0.38 474nm マンガン薄膜 黄 0.45 0.45 466 赤紫 0.44 0.36 490 チタン薄膜 黄 0.44 0.43 470 赤紫 0.42 0.33 494 ニッケル薄膜 黄 0.43 0.47 441 橙 0.46 0.41 467 銅薄膜 黄 0.48 0.44 459 黄 0.49 0.39 485 アルミ薄膜 黄 0.41 0.46 421 黄 0.44 0.48 441[Table 2] Before AN thin film deposition After AN thin film deposition Light-reflecting solid surface Color tone X coordinate Y coordinate Absorption wavelength Color tone X coordinate Y coordinate Absorption wavelength Chrome thin film Yellow 0.39 0.45 452nm Red-purple 0.41 0.38 474nm Manganese thin film Yellow 0.45 0.45 466 Red-purple 0.44 0.36 490 Titanium thin film Yellow 0.44 0.43 470 Magenta 0.42 0.33 494 Nickel thin film Yellow 0.43 0.47 441 Orange 0.46 0.41 467 Copper thin film Yellow 0.48 0.44 459 Yellow 0.49 0.39 485 Aluminum thin film Yellow 0.41 0.46 421 Yellow 0.44 0.48 441
【００２７】光反射性固体表面の屈折率がＡＮ薄膜着膜
による光学特性に及ぼす影響について、ＣＩＥ（国際照
明委員会）の色度図に基づき説明する。図３は、透明な
基板表面に薄膜を堆積させたとき、その干渉色がどの様
に変化するのかを色度図上で定性的に示すものである。
図のように、薄膜厚さの増加に伴い、その干渉色は図中
原点からの軌跡を矢印の方向に移動する。このとき、干
渉色の色調と膜厚の増加分とが同じであっても、原点か
らの軌跡の１周目と２周目とでは軌跡上の移動距離が異
なり、周回を重ねるほど移動距離は小さくなる。また、
同じ周回でかつ同じ膜厚増加であっても、原点に近い軌
跡をとるほど色調変化は大きくなるが、距離が小さすぎ
るときには軌跡が白色領域に入り込み、色調変化が検知
できなくなる。The influence of the refractive index of the light-reflecting solid surface on the optical properties of the AN thin film is described with reference to the chromaticity diagram of CIE (International Commission on Illumination). FIG. 3 qualitatively shows on the chromaticity diagram how the interference color changes when a thin film is deposited on the surface of a transparent substrate.
As shown in the figure, as the thickness of the thin film increases, the interference color moves along the locus from the origin in the direction of the arrow in the figure. At this time, even if the color tone of the interference color and the increment of the film thickness are the same, the moving distance on the locus differs between the first and second laps of the locus from the origin, and the more the laps, the more the moving distance becomes. Get smaller. Also,
Even if the trajectory is the same and the film thickness is the same, the change in color tone increases as the locus closer to the origin is taken, but if the distance is too small, the locus enters the white region and the change in color tone cannot be detected.
【００２８】上記した色度図上での一般的な干渉色の変
化を考慮すると、ＡＮ薄膜堆積による色度図上の軌跡
は、反射性固体表面の屈折率が小さいほど原点から離れ
た位置をとり、そのために色調変化としての検出感度が
低下するものと考えられる。また、薄膜の厚さの違いを
色調変化としても最も鋭敏に感じられるのは赤から紫の
領域であることから、ＡＮ薄膜を堆積する前の色調は、
色度軌跡で赤の前になる黄色から赤色までの領域に揃え
る必要がある。Considering the general change of the interference color on the chromaticity diagram described above, the locus on the chromaticity diagram due to the AN thin film deposition is located at a position farther from the origin as the refractive index of the reflective solid surface is smaller. Therefore, it is considered that the detection sensitivity as a change in color tone is lowered due to this. Further, even if the difference in the thickness of the thin film is considered as the change in color tone, it is the region from red to purple that is most sensitively sensed, so the color tone before depositing the AN thin film is
It is necessary to align the area from yellow to red, which is in front of red in the chromaticity locus.
【００２９】従来技術の中には、本発明と同様に光反射
性固体の表面に光干渉層を設けることが開示されてお
り、その光反射性固体の材料として本発明によるクロム
を含む種々の金属が上げられている。しかし、従来技術
ではその反射性に注目しているのみで、本発明になる如
く光反射性固体表面とその上に堆積される光干渉層との
屈折率の違いは何等検討されていない。そのため、従来
技術では単なる定性的な検出に留まったものである。The prior art discloses that a light interference layer is provided on the surface of a light-reflective solid as in the present invention, and various materials containing chromium according to the present invention as a material of the light-reflective solid are disclosed. The metal is raised. However, the prior art only pays attention to its reflectivity, and no consideration has been made on the difference in refractive index between the light-reflective solid surface and the light interference layer deposited thereon as in the present invention. Therefore, the conventional technique is limited to qualitative detection.
【００３０】以上の実験結果を基に、ＡＮ薄膜堆積前の
干渉色を黄色〜橙色の領域に制御するため、光干渉層の
構成について検討した。表３は、クロム薄膜を蒸着した
スライドガラスを光反射性固体表面とし、その上に設け
たシリコン薄膜の厚みと光学特性との関係を示すもので
ある。このとき、酸化シリコン薄膜の厚みは１２０nm、
また二酸化シリコンの厚みは２０nmとした。Based on the above experimental results, the structure of the optical interference layer was examined in order to control the interference color before the deposition of the AN thin film in the yellow to orange region. Table 3 shows the relationship between the optical characteristics and the thickness of the silicon thin film provided on the slide glass on which the chromium thin film is vapor-deposited as the light-reflecting solid surface. At this time, the thickness of the silicon oxide thin film is 120 nm,
The thickness of silicon dioxide was 20 nm.
【００３１】[0031]
【表３】 ＡＮ薄膜着膜前 ＡＮ薄膜着膜後 シリコン膜厚 色調 Ｘ座標 Ｙ座標 吸収波長 色調 Ｘ座標 Ｙ座標 吸収波長 75 nm 黄 0.38 0.50 448nm 濁橙 0.40 0.41 480nm 150 薄紫 0.26 0.22 518 紫 0.24 0.21 533 340 黄 0.40 0.46 436 橙 0.47 0.43 475 490 黄 0.40 0.50 429 橙 0.47 0.44 466 560 黄 0.44 0.45 456 桃 0.47 0.42 475[Table 3] Before AN thin film deposition After AN thin film deposition Silicon film thickness Color tone X coordinate Y coordinate Absorption wavelength Color tone X coordinate Y coordinate Absorption wavelength 75 nm Yellow 0.38 0.50 448 nm Turbid orange 0.40 0.41 480 nm 150 Light purple 0.26 0.22 518 Purple 0.24 0.21 533 340 Yellow 0.40 0.46 436 Orange 0.47 0.43 475 490 Yellow 0.40 0.50 429 Orange 0.47 0.44 466 560 Yellow 0.44 0.45 456 Pink 0.47 0.42 475
【００３２】表３のように、実験したシリコン膜厚の範
囲内では、その厚みが大きいほど明瞭な色調変化を示し
た。これは、膜厚が小さいときには可視光の吸収が散漫
になり、下地にあるクロム薄膜の色調の影響を受けるた
めである。このことから、光干渉層としてのシリコンの
膜厚は３４０nm以上であることが好ましい。しかし、厚
すぎる場合には前記したシリコンウェハーのときと同様
の問題を生じるので、３４０〜５６０ｎｍ以下であるこ
とが望ましい。As shown in Table 3, within the range of the experimental silicon film thickness, the larger the thickness, the more obvious the change in color tone. This is because when the film thickness is small, the absorption of visible light is diffused and is affected by the color tone of the underlying chromium thin film. From this, it is preferable that the film thickness of silicon as the light interference layer is 340 nm or more. However, if it is too thick, the same problems as in the case of the above-mentioned silicon wafer occur, so that the thickness is preferably 340 to 560 nm or less.
【００３３】次に、光干渉層の中間領域を構成する酸化
シリコンの厚みについて、前記同様に検討した。このと
き、シリコン膜厚は３９０nm、また二酸化シリコンは２
０nmとした。得られた結果は表４の通りであり、酸化シ
リコン薄膜が厚いほど色度図の軌跡上ではより進んだ位
置を占めることになる。また、膜厚が２５nmと１３０nm
とを比べると、ＡＮ薄膜堆積前は同じ黄色を呈している
が、ＡＮ薄膜堆積による色調変化は２５nmの方が鮮明で
あった。これは、前記した軌跡が１周目と２周目との違
いによるものであり、鮮明な色調変化を得るには酸化シ
リコン薄膜の厚みが２５nm〜１３０nmの範囲より好まし
くは２５nm〜５０nmの範囲にあることが望まれる。Next, the thickness of silicon oxide forming the intermediate region of the optical interference layer was examined in the same manner as above. At this time, the silicon film thickness is 390 nm and the silicon dioxide is 2
It was set to 0 nm. The obtained results are shown in Table 4, and the thicker the silicon oxide thin film, the more advanced the position on the locus of the chromaticity diagram. Also, the film thickness is 25 nm and 130 nm
Comparing with the above, the same yellow color was exhibited before the AN thin film was deposited, but the change in color tone due to the AN thin film deposition was clearer at 25 nm. This is because the above-mentioned locus is due to the difference between the first lap and the second lap, and in order to obtain a clear color tone change, the thickness of the silicon oxide thin film is preferably in the range of 25 nm to 130 nm, more preferably in the range of 25 nm to 50 nm. Is desired.
【００３４】[0034]
【表４】 ＡＮ薄膜着膜前 ＡＮ薄膜着膜後 酸化シリコン膜厚 色調 Ｘ座標 Ｙ座標 吸収波長 色調 Ｘ座標 Ｙ座標 吸収波長 25 nm 黄 0.40 0.39 390nm 赤 0.45 0.31 501nm 50 赤紫 0.39 0.20 525 青 0.22 0.21 - 72 黄 0.41 0.46 - 黄橙 0.46 0.45 - 130 黄 0.41 0.46 431 黄橙 0.46 0.45 458 160 橙桃 0.43 0.36 482 紫桃 0.38 0.25 511[Table 4] Before AN thin film deposition After AN thin film deposition Silicon oxide film thickness Color tone X coordinate Y coordinate Absorption wavelength Color tone X coordinate Y coordinate Absorption wavelength 25 nm yellow 0.40 0.39 390 nm red 0.45 0.31 501 nm 50 Magenta 0.39 0.20 525 Blue 0.22 0.21- 72 Yellow 0.41 0.46-Yellow Orange 0.46 0.45- 130 Yellow 0.41 0.46 431 Yellow Orange 0.46 0.45 458 160 Orange peach 0.43 0.36 482 Purple peach 0.38 0.25 511
【００３５】光干渉層の最上部を構成する二酸化シリコ
ンの厚みについても、上記と同様に調べた。このとき、
シリコン膜厚は４４０nm、また酸化シリコン膜厚は２５
nmとした。得られた結果を表５に示す。表のように、二
酸化シリコン薄膜が厚いほど色調が白色の領域に入り、
変化を検出するには充分でなかった。すなわち、二酸化
シリコンの厚みは軌跡の原点からの距離を変化させ、厚
みが大きいほどその距離は小さくなるため、その厚みは
１５nm〜６０nm、より好ましくは１５nm〜４０nmに制御
する必要がある。The thickness of silicon dioxide forming the uppermost part of the light interference layer was also examined in the same manner as above. At this time,
Silicon film thickness is 440 nm and silicon oxide film thickness is 25
nm. The results obtained are shown in Table 5. As shown in the table, the thicker the silicon dioxide thin film, the more the color tone enters the white area,
It was not enough to detect the change. That is, the thickness of silicon dioxide changes the distance from the origin of the locus, and the larger the thickness, the smaller the distance. Therefore, it is necessary to control the thickness to 15 nm to 60 nm, more preferably 15 nm to 40 nm.
【００３６】[0036]
【表５】 ＡＮ薄膜着膜前 ＡＮ薄膜着膜後 二酸化シリコン膜厚 色調 Ｘ座標 Ｙ座標 吸収波長 色調 Ｘ座標 Ｙ座標 吸収波長 15 nm 淡黄 0.35 0.37 365nm 赤 0.48 0.41 450nm 20 淡茶 0.37 0.38 384 赤紫 0.46 0.33 496 40 茶 0.38 0.39 399 青紫 0.34 0.23 496 60 赤茶 0.42 0.39 419 青 0.26 0.21 - 80 青茶 0.41 0.35 476 青 0.24 0.23 -[Table 5] Before AN thin film deposition After AN thin film deposition Silicon dioxide film thickness Color tone X coordinate Y coordinate Absorption wavelength Color tone X coordinate Y coordinate Absorption wavelength 15 nm Light yellow 0.35 0.37 365nm Red 0.48 0.41 450nm 20 Light brown 0.37 0.38 384 Red purple 0.46 0.33 496 40 Brown 0.38 0.39 399 Blue Purple 0.34 0.23 496 60 Red Brown 0.42 0.39 419 Blue 0.26 0.21- 80 Blue Brown 0.41 0.35 476 Blue 0.24 0.23-
【００３７】以上の結果から、光干渉層はその表面に近
づくほど屈折率が小さくなるように構成し、その下地に
なる光反射性固体表面は光干渉層の表面よりも大きな屈
折率を有する材料とし、かつ有機物質を堆積させる前の
光学特性が、色度図の座標上ではＸ軸が０．３５から
０．４５でＹ軸が０．３７から０．５０の領域にあり、
反射分光特性の吸収波長が３６５nmから５２５nmの範囲
に制御したとき、光学的免疫測定に供するデバイスとし
ての感度が最も大きくなる。From the above results, the light interference layer is constructed so that its refractive index becomes smaller as it gets closer to the surface thereof, and the underlying light-reflective solid surface is a material having a larger refractive index than the surface of the light interference layer. And the optical characteristics before depositing the organic substance are in the region of X-axis 0.35 to 0.45 and Y-axis 0.37 to 0.50 on the coordinates of the chromaticity diagram,
When the absorption wavelength of the reflection spectral characteristic is controlled in the range of 365 nm to 525 nm, the sensitivity as a device used for the optical immunoassay becomes maximum.
【００３８】これらの実験結果に基づき、図４に示すよ
うに、スライドガラス１１の表面にクロム薄膜１２を設
けた光反射性固体表面に、厚みが４００nmのシリコン薄
膜１３、２５nmの一酸化シリコン薄膜１４および２０nm
の二酸化シリコン薄膜１５を順次堆積させ、次にプラズ
マ中への導入ガスをアリルアミン（以下、ＡＡとする）
モノマーに切り替え、約４nmのＡＡ薄膜１６を形成し
た。ここで、光干渉層の最表層にＡＡ薄膜を設けたの
は、光干渉層の表面に有機物質の吸着能を持たせるため
であり、二酸化シリコンとＡＡ薄膜との屈折率がほぼ等
しいことを考慮すると、ＡＡ薄膜の下層にある二酸化シ
リコン薄膜の全てをＡＡ薄膜に置き換えることも可能で
ある。Based on these experimental results, as shown in FIG. 4, a 400 nm thick silicon thin film 13 and a 25 nm silicon monoxide thin film were formed on a light-reflecting solid surface having a chromium thin film 12 provided on the surface of a slide glass 11. 14 and 20 nm
A silicon dioxide thin film 15 is sequentially deposited, and then allylamine (hereinafter referred to as AA) is used as a gas to be introduced into the plasma.
By switching to a monomer, an AA thin film 16 of about 4 nm was formed. Here, the reason why the AA thin film is provided on the outermost surface of the light interference layer is to give the surface of the light interference layer the ability to adsorb an organic substance, and that the refractive index of silicon dioxide and that of the AA thin film are almost the same. Considering that, it is possible to replace all of the silicon dioxide thin film under the AA thin film with the AA thin film.
【００３９】この多層構造基板の表面に、濃度が１ng／
mlから５mg／mlの領域で種々の濃度に調整したグルコー
スオキシターゼ（以下、ＧＯＤとする）を抗原として滴
下して４℃で一晩放置した後、蒸留水で洗浄したものを
光学的免疫測定デバイスとした。このときのデバイスの
色調はＧＯＤを滴下する前と同じ赤茶色を呈し、またＧ
ＯＤの濃度による違いも認められず、他の光学特性も殆
ど変化しなかった。ここで、滴下された高濃度のＧＯＤ
がＡＡ薄膜の表面に単分子層を形成するなら、それは前
記の実験でＡＮ薄膜を堆積したのと等価であり、色調は
赤茶色から紫色に変化するはずである。しかし、本実験
でＧＯＤ滴下による色調変化が認められなかったのは、
ＧＯＤが単分子層を形成するのではなく、きわめて低密
度に吸着されていることを意味している。On the surface of this multilayer structure substrate, a concentration of 1 ng /
Glucose oxidase (hereinafter referred to as GOD) adjusted to various concentrations in the range of 5 to 5 mg / ml was added dropwise as an antigen, left at 4 ° C overnight, and washed with distilled water. And The color tone of the device at this time is the same reddish brown as before GOD was dropped, and G
No difference was observed depending on the OD concentration, and other optical characteristics were hardly changed. Here, the high concentration of GOD dropped
If forms a monolayer on the surface of the AA thin film, it is equivalent to depositing the AN thin film in the above experiment and the color should change from reddish brown to purple. However, in this experiment, the change in color tone due to GOD dropping was not observed.
It means that the GOD is adsorbed at an extremely low density rather than forming a monolayer.
【００４０】この抗原としてのＧＯＤが低密度に吸着さ
れた光学的免疫測定デバイスに、ウサギ由来抗ＧＯＤポ
リクロナール抗体を滴下し、４℃で一晩放置することに
より抗原抗体反応を生じせしめた。その後、蒸留水で洗
浄して乾燥させ、白色光下で目視観察した結果、ＧＯＤ
を滴下しなかったものでは色調に何等の変化もなかった
が、ＧＯＤが低密度に吸着されたデバイスでは色調変化
が認められ、その濃淡は滴下したＧＯＤ濃度に応じて異
なった。表６は、この一連の実験結果を示したものであ
る。A rabbit-derived anti-GOD polyclonal antibody was dropped onto the optical immunoassay device in which GOD as an antigen was adsorbed at a low density, and the mixture was allowed to stand overnight at 4 ° C. to cause an antigen-antibody reaction. Then, it was washed with distilled water, dried, and visually observed under white light.
The color tone did not change at all in the case of not dropping, but the color tone change was observed in the device in which GOD was adsorbed at a low density, and the shade was different depending on the dropped GOD concentration. Table 6 shows the results of this series of experiments.
【００４１】[0041]
【表６】 抗原抗体反応前 抗原抗体反応後 ＧＯＤ濃度 色調 Ｘ座標 Ｙ座標 吸収波長 色調 Ｘ座標 Ｙ座標 吸収波長 5 mg/ml 赤茶 0.41 0.41 396nm 赤桃 0.40 0.30 516nm 1 赤茶 同上 同上 深赤紫 0.41 0.33 487 0.1 赤茶 同上 同上 濃赤紫 0.42 0.34 475 0.01 赤茶 同上 同上 赤紫 0.43 0.37 465 0.001 赤茶 同上 同上 薄赤紫 0.43 0.36 466[Table 6] Before antigen-antibody reaction After antigen-antibody reaction GOD density Color tone X coordinate Y coordinate Absorption wavelength Color tone X coordinate Y coordinate Absorption wavelength 5 mg / ml red brown 0.41 0.41 396nm red peach 0.40 0.30 516nm 1 Red brown Same as above Same as deep red purple 0.41 0.33 487 0.1 Red brown Same as above Same as above Deep red-purple 0.42 0.34 475 0.01 Red brown Same as above Same as red purple 0.43 0.37 465 0.001 Red brown Same as above Same as light red purple 0.43 0.36 466
【００４２】表６に示したように、本発明による光学的
免疫測定デバイスは、抗原の濃度が１ng／mlでも目視に
よる色調変化として捉え得ることが分かった。また、色
調や色度の変化から、ＧＯＤが５mg／mlと高濃度の場合
でも抗原抗体反応による有機物質の厚みの増加が、その
単分子としての厚みには達していないことが分かった。
すなわち、この事実はデバイス表面に吸着された有機物
質の密度の増加を色調変化として捉えているものであ
り、さらに有機物質の密度を増加させることにより検出
感度の増大が期待された。As shown in Table 6, it was found that the optical immunoassay device of the present invention can be visually recognized as a change in color tone even when the antigen concentration is 1 ng / ml. Further, from the change in color tone and chromaticity, it was found that the increase in the thickness of the organic substance due to the antigen-antibody reaction did not reach the thickness as a single molecule even when GOD was as high as 5 mg / ml.
That is, this fact captures an increase in the density of the organic substance adsorbed on the device surface as a change in color tone, and further increase in the density of the organic substance was expected to increase the detection sensitivity.
【００４３】そこで、濃度が０．１ng／mlから１mg／ml
の範囲で、種々の濃度のＧＯＤを滴下、洗浄および乾燥
させたデバイスに、前記と同様にウサギ由来抗ＧＯＤポ
リクロナール抗体を滴下して洗浄、乾燥させた後、さら
にヒツジ由来抗（ウサギＩgＧ）抗体を滴下して４℃で
一晩放置することによりウサギ由来抗ＧＯＤポリクロナ
ール抗体と反応させ、蒸留水で洗浄して乾燥し、前の実
験と同様に白色光下で目視観察した。その結果は表７に
示すように、滴下したＧＯＤ濃度が１mg／mlのデバイス
の色調は深い赤紫へと大きく変化し、ヒツジ由来抗（ウ
サギＩgＧ）抗体を滴下する前には色調変化が認められ
なかったＧＯＤ濃度（０．１ng／ml）においても変化が
検出された。Therefore, the concentration is from 0.1 ng / ml to 1 mg / ml.
In the same manner as described above, a rabbit-derived anti-GOD polyclonal antibody was added dropwise to a device in which various concentrations of GOD were dropped, washed, and dried, and then washed and dried, and then sheep-derived anti- (rabbit IgG) antibody was added. Was reacted with the rabbit-derived anti-GOD polyclonal antibody by allowing it to stand overnight at 4 ° C., washed with distilled water, dried, and visually observed under white light as in the previous experiment. As a result, as shown in Table 7, the color tone of the device with the dropped GOD concentration of 1 mg / ml changed greatly to deep red purple, and the color tone change was observed before the sheep-derived anti- (rabbit IgG) antibody was dropped. Changes were also detected in GOD concentrations that were not (0.1 ng / ml).
【００４４】[0044]
【表７】 抗原抗体反応前 抗原抗体反応後 ＧＯＤ濃度 色調 Ｘ座標 Ｙ座標 吸収波長 色調 Ｘ座標 Ｙ座標 吸収波長 1 mg/ml 赤茶 0.39 0.40 - nm 深赤紫 0.41 0.40 463nm 0.1 赤茶 同上 同上 濃赤紫 0.41 0.41 449 0.01 赤茶 同上 同上 赤紫 0.40 0.41 428 0.001 赤茶 同上 同上 薄赤紫 0.41 0.41 420 0.0001 赤茶 同上 同上 淡赤紫 0.40 0.41 -[Table 7] Before antigen-antibody reaction After antigen-antibody reaction GOD density Color tone X coordinate Y coordinate Absorption wavelength Color tone X coordinate Y coordinate Absorption wavelength 1 mg / ml red brown 0.39 0.40-nm deep red purple 0.41 0.40 463nm 0.1 Red brown Same as above Same as deep red purple 0.41 0.41 449 0.01 Red Brown Same as above Same as Red Purple 0.40 0.41 428 0.001 Red brown Same as above Same as light reddish purple 0.41 0.41 420 0.0001 Red brown Same as above Same as light red purple 0.40 0.41-
【００４５】以上、一連の実験結果は、光干渉層となる
多層構造表面に抗原を低密度に吸着させ、そこで抗原抗
体反応を生じせしめることにより、その密度のわずかな
増加を光学的膜厚の増加として色調変化により定量的に
捉え得ることを示した。このことから、抗原ではなく抗
体を低密度に吸着させても同様の効果が得られると考え
られる。The results of a series of experiments described above show that the antigen is adsorbed at a low density on the surface of the multi-layer structure serving as the light interference layer, and an antigen-antibody reaction is caused there to cause a slight increase in the density to reduce the optical film thickness. It was shown that the increase can be quantitatively grasped by the change in color tone. From this, it is considered that the same effect can be obtained by adsorbing the antibody instead of the antigen at a low density.
【００４６】一方、本発明では多層構造表面に抗原を吸
着させたが、例えば多層構造の表層を製膜するとき、Ａ
Ａモノマーに変えてアクリル酸モノマーやアセトアルデ
ヒドモノマーを製膜チャンバーに導入し、その表層にカ
ルボキシル基やアルデヒド基を導入することにより、抗
原や抗体を低密度に化学結合させることも可能となる。On the other hand, in the present invention, the antigen was adsorbed on the surface of the multilayer structure.
By introducing an acrylic acid monomer or an acetaldehyde monomer into the film forming chamber instead of the A monomer and introducing a carboxyl group or an aldehyde group into the surface layer, it is possible to chemically bond the antigen or antibody at a low density.
【００４７】[0047]
【効果】光反射性固体表面および光干渉層になる多層構
造と光学特性について詳細に検討した結果、抗原抗体反
応により表面で生じる有機物質のわずかな密度増加を目
視による色調変化として定量的に捉え得ることが可能に
なり、きわめて簡便な光学的免疫測定法が提供できた。[Effect] As a result of detailed examination of the multilayer structure and the optical properties of the light-reflecting solid surface and the light interference layer, a slight increase in the density of organic substances generated on the surface due to the antigen-antibody reaction was quantitatively captured as a visual color change. It has become possible to provide an extremely simple optical immunoassay method.
【００４８】上記免疫測定に供するデバイスについて、
従来はＬＢ法のような実験室的な手法に頼っていた光干
渉層の多層構造を、量産技術として実績のあるプラズマ
反応法で連続的に形成可能としたことから、そのデバイ
スが安価でかつ大量に供給し得るようになった。Regarding the device used for the above-mentioned immunoassay,
Since the multilayer structure of the optical interference layer, which has conventionally relied on a laboratory method such as the LB method, can be continuously formed by the plasma reaction method, which has a proven track record as a mass production technology, the device is inexpensive and It became possible to supply in large quantities.
【００４９】[0049]
【図１】 光干渉層になる多層構造を形成する製膜装置
の概念図である。FIG. 1 is a conceptual diagram of a film forming apparatus that forms a multilayer structure that serves as an optical interference layer.
【図２】 図中（ａ）は本発明にかかるデバイス構造の
断面図、（ｂ）は比較例１の断面図、（ｃ）は比較例２
の断面図である。2A is a sectional view of a device structure according to the present invention, FIG. 2B is a sectional view of Comparative Example 1, and FIG. 2C is Comparative Example 2.
FIG.
【図３】 一般的な干渉色と薄膜の膜厚との関係を示す
ＣＩＥの色度図である。FIG. 3 is a CIE chromaticity diagram showing a relationship between a general interference color and a film thickness of a thin film.
【図４】 本発明にかかる光学的免疫測定用デバイスの
断面図である。FIG. 4 is a cross-sectional view of a device for optical immunoassay according to the present invention.
１１…スライドグラス、 １２…クロム薄膜、 １３…シリコン薄膜、 １４…酸化シリコン薄膜、 １５…二酸化シリコン薄膜、 １６…ＡＡ薄膜。 11 ... slide glass, 12 ... Chrome thin film, 13 ... Silicon thin film, 14 ... Silicon oxide thin film, 15 ... Silicon dioxide thin film, 16 ... AA thin film.
───────────────────────────────────────────────────── フロントページの続き (56)参考文献 特開 平５−129277（ＪＰ，Ａ) 特公 平４−8742（ＪＰ，Ｂ２) 特表 平５−506936（ＪＰ，Ａ) (58)調査した分野(Int.Cl.7，ＤＢ名) G01N 33/543 595 G01N 21/45 ─────────────────────────────────────────────────── ─── Continuation of the front page (56) References JP-A-5-129277 (JP, A) JP-B-4-8742 (JP, B2) JP-A-5-506936 (JP, A) (58) Field (Int.Cl. 7 , DB name) G01N 33/543 595 G01N 21/45
Claims (3)
くほど小さくなる多層構造を有すると共に、可視光を吸
収する光干渉層を形成して、その表面に抗原又は抗体の
いずれか一方である第１の有機物質を予め単分子層を形
成する分子数以下の量を吸着させておき、吸着させた前
記第１の有機物質と抗原抗体反応を生ずる抗体又は抗原
のいずれかである第２の有機物質を与えて抗原抗体反応
を生じさせ、その抗原抗体反応の結果生ずる前記光干渉
層表面での有機物質密度の増加を可視光領域での光干渉
色の変化として定量的に検出することを特徴とする光学
的免疫測定法。1. A light-reflecting layer having a refractive index close to that of its surface.
It has a multi-layer structure that becomes smaller and smaller, and forms a light interference layer that absorbs visible light, and forms a monomolecular layer on the surface of the first organic substance that is either an antigen or an antibody in advance.
An amount of molecules or less to be adsorbed, and a second organic substance that is either an antibody or an antigen that causes an antigen-antibody reaction with the adsorbed first organic substance is given to give an antigen-antibody cause reaction, optical immunoassay, characterized in that quantitatively detect an increase in the organic matter density at the optical interference layer surface caused a result of the antigen-antibody reaction as a change in optical interference color in the visible light region Law.
に近づくほど小さくなる多層構造を有すると共に、可視
光が吸収されかつ有機物質が吸着される表面を有する光
干渉層を設け、前記光反射性固体表面は前記光干渉層表
面よりも大きな屈折率を有する材料で構成し、前記光干
渉層の表面に単分子層を形成する分子数以下の量の抗原
もしくは抗体を吸着させたことを特徴とする光学的免疫
測定デバイス。2. A light-reflecting solid surface having a refractive index
A light interference layer having a surface that absorbs visible light and adsorbs an organic substance is provided with a multi-layer structure that becomes smaller as it gets closer to, and the light-reflecting solid surface has a larger refractive index than the light interference layer surface. An optical immunoassay device, which is made of a material having the above and has adsorbed an amount of antigen or antibody equal to or less than the number of molecules forming a monomolecular layer on the surface of the light interference layer.
される請求項２に記載の光学的免疫測定デバイス。3. The optical immunoassay device according to claim 2, wherein the light interference layer is formed by a plasma reaction method.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP32852893A JP3455262B2 (en) | 1993-12-24 | 1993-12-24 | Optical immunoassay and device |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP32852893A JP3455262B2 (en) | 1993-12-24 | 1993-12-24 | Optical immunoassay and device |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH07181185A JPH07181185A (en) | 1995-07-21 |
| JP3455262B2 true JP3455262B2 (en) | 2003-10-14 |
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ID=18211301
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP32852893A Expired - Fee Related JP3455262B2 (en) | 1993-12-24 | 1993-12-24 | Optical immunoassay and device |
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| Country | Link |
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| JP (1) | JP3455262B2 (en) |
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| RU2490616C2 (en) * | 2009-05-22 | 2013-08-20 | 3М Инновейтив Пропертиз Компани | Multilayer colour sensors |
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1993
- 1993-12-24 JP JP32852893A patent/JP3455262B2/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| JPH07181185A (en) | 1995-07-21 |
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