JPS62501933A - Fluorescence analysis arrangement - Google Patents
Fluorescence analysis arrangementInfo
- Publication number
- JPS62501933A JPS62501933A JP61501506A JP50150686A JPS62501933A JP S62501933 A JPS62501933 A JP S62501933A JP 61501506 A JP61501506 A JP 61501506A JP 50150686 A JP50150686 A JP 50150686A JP S62501933 A JPS62501933 A JP S62501933A
- Authority
- JP
- Japan
- Prior art keywords
- sample
- fluorescent
- analysis arrangement
- light
- incident light
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
Landscapes
- Health & Medical Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Physics & Mathematics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
- Investigating Or Analysing Materials By Optical Means (AREA)
Abstract
(57)【要約】本公報は電子出願前の出願データであるため要約のデータは記録されません。 (57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.
Description
【発明の詳細な説明】 螢光分析配置 本発明は螢光分析配置、特に螢光免疫測定に適用される螢光分析配置に関するも のである。[Detailed description of the invention] Fluorescence analysis arrangement The present invention also relates to a fluorescence analysis arrangement, in particular a fluorescence analysis arrangement applied to fluorescence immunoassays. It is.
螢光標識抗体をトレーサとして用いる技術は免疫アッセイ(分析)システムにお いては確立されているが、残留螢光標識抗体即ち蛋白質の確認は螢光顕微鏡検査 により行い、従って使用者のカラーおよび空間肉眼識別能力に臨界的に依存する 。無螢光標識蛋白質から発生する背景螢光を識別する時間分解螢光免疫測定法が 最近導入され、これにより定蛍処理を行うが、この目的のため市販されている螢 光光度計は液状媒体のみを処理するには好適であるが、紙又はプラスチック基板 上の多重サンプルを試験するには好適でない。高速螢光免疫測定法に対する良好 なアナログアプローチは、かかる基板、代表的には膜構体とし得る基板に残留螢 光標識物質を結合することである。The technology of using fluorescently labeled antibodies as tracers can be used in immunoassay (analysis) systems. Although it is well established that residual fluorescently labeled antibodies or proteins can be detected by fluorescence microscopy, and is therefore critically dependent on the user's color and spatial visual discrimination abilities. . A time-resolved fluorescence immunoassay method that identifies background fluorescence generated from nonfluorescently labeled proteins is now available. Recently introduced, this provides a constant fluorescing treatment, but the fluorophores that are commercially available for this purpose are Photometers are suitable for processing liquid media only, but with paper or plastic substrates. Not suitable for testing multiple samples above. Good for fast fluorescence immunoassay A typical analog approach is to remove residual fluorescent light from such a substrate, which typically can be a membrane structure. It involves binding a photolabeling substance.
基板、特に紙製の基板上のサンプルを定蛍測定する際の困難性は螢光から識別す る必要のある反射入射光の割合が高いことである。紙は螢光から期待される角度 分布と同一の角度分布で光を拡散反射する傾向がある。The difficulty in measuring constant fluorescence of samples on substrates, especially paper substrates, is that they cannot be distinguished from the fluorescence. The problem is that the proportion of reflected incident light that needs to be filtered is high. The paper is at the angle expected from the fluorescence. There is a tendency to diffusely reflect light with the same angular distribution.
本発明の目的は、斯かる困難性の欠点を少なくとも部分的に除去し得るように構 成した螢光分析配置を提供せんとするにある。The object of the present invention is to provide a structure in such a way that the drawback of such difficulty can be at least partially obviated. The aim is to provide a complete fluorescence analysis arrangement.
本発明螢光分析配置は、サンプル支持体と、入射光の光源と、サンプルから放出 された螢光の検出器と、サンプルから反射された入射光から螢光を分離するフィ ルタ手段を位置決めする手段とを備えることを特徴とする。The fluorescence analysis arrangement of the present invention comprises a sample support, a source of incident light, and a sample support for emitted light from the sample. a detector for the reflected fluorescence and a filter that separates the fluorescence from the incident light reflected from the sample. and means for positioning the router means.
代表的な場合には螢光を、入射光よりも高い波長とすると共にフィルタ手段はグ イクロイックフィルタを具え得るようにする。又、フィルタ手段は、入射光およ びその反射光を透過し、且つ螢光を入射光に対しを測角で反射するように選択さ れた帯域通過干渉フィルタとすることができる。Typically, the fluorescent light will be of a higher wavelength than the incident light and the filter means will be of a higher wavelength. Equipped with an ichroic filter. Further, the filter means filters the incident light and The fluorescent light is selected so that it transmits the reflected light and reflects the fluorescent light at an angle relative to the incident light. It can be a bandpass interference filter.
帯域通過干渉フィルタは、これを反射された入射光に対し適宜配列して入射光の フィルタへの最小入射角が螢光の好適且つ実際の角度分離に一致し得るようにす るのが好適である。かかる入射角は10度〜20度の範囲とする。Band-pass interference filters are arranged appropriately for the reflected incident light to filter out the incident light. Ensure that the minimum angle of incidence on the filter corresponds to the preferred and actual angular separation of the fluorescent light. It is preferable to Such an angle of incidence is in the range of 10 degrees to 20 degrees.
又、フィルタ手段は螢光分析配列の一部分を具えるようにする。The filter means may also comprise part of the fluorometric array.
更に本発明螢光分析配置の他の例は基準サンプルおよび試験サンプルを支持する 手段と、入射光の光源と、サンプルから放出された螢光の検出器と、入射光を基 準サンプル又は試験サンプルに選択的に向けると共に個別の蛍光を検出器に向け るように調整し得る光学手段とを具えることを特徴とする。Further examples of the present invention fluorometric analysis arrangement support reference samples and test samples. means, a source of incident light, a detector of fluorescence emitted from the sample, and a source of incident light; Selectively direct the quasi-sample or test sample and direct individual fluorescence to the detector and an optical means that can be adjusted to
上述した調整可能な光学手段は入射光を基準サンプルおよび試験サンプルに交互 且つ周期的に向:すると共に個別の螢光を受けて検出器に向けるために振動する ように配設された枢動自在のミラーを具えるようにする。かかるミラーは、2つ のサンプルと、各サンプルにより反射された入射光から螢光を分離するフィルタ 手段との間の光学路に配列するのが好適である。The adjustable optical means described above alternately directs the incident light to the reference sample and the test sample. and periodically vibrate to receive individual fluorescent light and direct it to the detector. A pivotable mirror is provided. There are two mirrors samples and a filter that separates the fluorescent light from the incident light reflected by each sample. Preferably, it is arranged in an optical path between the means.
本発明では個別の光路を画成する好適な光学装置を設ける。かかる装置は、代表 的には1個以上のレンズと、螢光検出器に関連するフィルタとを設ける。又、位 相感応装置を設けて検出された信号を増強し得るようにする。この信号増強は、 例えば1例では入射光をパルス化すること即ぢ機械的に断続することにより、他 の例ではサンプルと検出器との間に光路が存在しない条件でミラーを振動させる ことにより行うことができる。The present invention provides a suitable optical arrangement for defining separate optical paths. Such equipment is representative Typically, one or more lenses and a filter associated with the fluorescence detector are provided. Also, rank A phase sensitive device may be provided to enhance the detected signal. This signal enhancement is For example, in one case, by pulsing the incident light, i.e. by mechanically intermittent In this example, the mirror is vibrated with no optical path between the sample and the detector. This can be done by
以下図面により本発明を説明する。The present invention will be explained below with reference to the drawings.
第1図は本発明による螢光分析配置の構成を示す説明図である。FIG. 1 is an explanatory diagram showing the configuration of a fluorescence analysis arrangement according to the present invention.
第1図に示す本発明螢光分析配置は、入射光5のランプル支持体14および15 と、光起電型検出器16と、ランプ光源12からサンプル10.11への入射光 の光路およびサンプル10゜11から検出器16への螢光の光路を交互に決める 1組の光学装置18とを具える。光学装置18は回転自在の振動ミラー20を具 え、これにより入射光5を基準サンプル10又は試験サンプル11に選択的に向 ける作用と、位相感応検出のために入射光および螢光を断続する作用との2つの 作用を呈し得るようにする。更に光学装置18は反射入射光から螢光6を分離す るビームスプリッタとして作用するグイクロイックフィルタ22と、このフィル タ22の近くの種々の光ビームを平行光とするように配列された3個のレンズ2 4.25および26と、検出器16の前面に設けられた検出増強分析フィルタ2 8とを具える。The fluorescent analysis arrangement of the invention shown in FIG. , a photovoltaic detector 16, and light incident on the sample 10.11 from the lamp light source 12. and the optical path of the fluorescent light from the sample 10° 11 to the detector 16 are determined alternately. a set of optical devices 18. The optical device 18 includes a rotatable vibrating mirror 20. This allows the incident light 5 to be selectively directed towards the reference sample 10 or the test sample 11. There are two functions: one to intermittent the incident light and the fluorescent light for phase-sensitive detection. To be able to exhibit an effect. Furthermore, the optical device 18 separates the fluorescent light 6 from the reflected incident light. a guichroic filter 22 that acts as a beam splitter; Three lenses 2 arranged to parallelize various light beams near the lens 22 4.25 and 26 and the detection enhancement analysis filter 2 provided in front of the detector 16 8.
基阜サンプル10を紙yの基板に載置する場合にはフィルタ22の近くの光ビー ムを平行光として入出力ビームのスペクトルがオーバーランプするのを減少し得 るようにするのが好適である。透明基板例えばプラスチック基板に対しては入出 力ビームが成る程度オーバーラツプするのは許容し得るため、平行光とすること は臨界的ではない。3個のレンズの代わりに例えばレンズ25の領域に単一レン ズを設けることもできる。When placing the base sample 10 on the paper y substrate, the light beam near the filter 22 It is possible to reduce the overlamp of the input and output beam spectra by making the beam parallel light. It is preferable that the Input/output for transparent substrates such as plastic substrates It is acceptable for the power beams to overlap to some extent, so parallel beams should be used. is not critical. For example, a single lens in the area of lens 25 can be used instead of three lenses. It is also possible to provide a
グイクロイックフィルタ22は、入射光およびその反射光を透過し、螢光6を検 出器16にほぼ全部反射する帯域通過干渉フィルタを具える。このフィルタは測 定毎に交換し得ると共に取付台23に保持する。このフィルタ22を適宜位置決 めして反射入射光の入射角θ1が螢光の好適な分離および構成部品の実際の物理 的な配置と最小で両立し得る角度となるようにする。この好適な角度は10度と し、20度以上としないようにするのが好適である。The guichroic filter 22 transmits the incident light and its reflected light, and detects the fluorescent light 6. The output device 16 is equipped with a bandpass interference filter that reflects almost all of the reflection. This filter It can be replaced at regular intervals and is held on the mounting base 23. Position this filter 22 appropriately. The angle of incidence θ1 of the reflected incident light is determined by the preferred separation of the fluorescent light and the actual physics of the components. The angle should be the minimum that is compatible with the general layout. This suitable angle is 10 degrees. However, it is preferable that the temperature should not exceed 20 degrees.
位相感応検出に振動ミラーを用いる代わりに簡単な電動機駆動型の機械式チョッ パを用い、これをランプ光源12とフィルタ22との間に位置させるようにする 。An alternative to using an oscillating mirror for phase-sensitive detection is a simple motor-driven mechanical chopper. and place it between the lamp light source 12 and the filter 22. .
レンズ24.25.26は大形のプラスチック型のレンズとすることができる。The lenses 24, 25, 26 can be large plastic type lenses.
その理由は検出器で良好な像を形成すことが臨界的でtいからである。この場合 f/15コーン全体による光の捕捉は極めて満足するものであった。The reason is that it is critical to form a good image on the detector. in this case The light capture by the entire f/15 cone was very satisfactory.
紙製支持体上に形成される像の大きさは代表的には約15度の照射角で10mm x2mmとすることができる。The size of the image formed on the paper support is typically 10 mm at an illumination angle of about 15 degrees. x2mm.
佐 図示の螢光分析配置を代表的な螢光免疫測定装置に適用する場合の例を以下に示 す。免疫測定法に広く用いられる螢光トレーサを、一般にフルオレスセインと称 されるフルオレスセインイソテオシアン酸塩とする。このトレーサは500nm のすぐ下側にピークを有する吸収帯と、530nmのすぐ下側にピークを有する 螢光帯とを有する。検出増強分析フィルタ28としては、約520nm以上の光 を透過するショットグラス型0G530フイルタを用いた。Sa An example of applying the illustrated fluorescence analysis arrangement to a typical fluorescence immunoassay device is shown below. vinegar. A fluorescent tracer widely used in immunoassays is generally called fluorescein. fluorescein isoteocyanate. This tracer is 500nm an absorption band with a peak just below 530 nm and a peak just below 530 nm. It has a fluorescent band. The detection enhancement analysis filter 28 uses light of approximately 520 nm or more. A shot glass type 0G530 filter that transmits
グイクロイックフィルタ22として選定した帯域通過干渉フィルタにはI F V 2型のフィルタを用いた。このフィルタは約430nmおよび470nm間 の狭い帯域に亘る透過度を有し、従って検出された信号内の反射光を減少するた めに著しく有利であった。The bandpass interference filter selected as the guichroic filter 22 has an IF A V2 type filter was used. This filter is between approximately 430nm and 470nm transmittance over a narrow band, thus reducing reflected light in the detected signal. It was extremely advantageous.
この場合の理想的な帯域通過干渉フィルタはその透過度を460nmないし48 0nmの範囲を中心として、500口m以上の長い波長の透過度は無視し得るよ うにする。又従来の波長堆積型フィルタ以外の“方形′°帯域通過フィルタ又は 誘起透過形フィルタを用いて所望の帯域除去を行うこともできる。更に、2個の 通常の同一のフィルタを並列に用いて帯域除去比を改善することもできる。フィ ルタの外面を被覆しない場合には反射性入力光のほぼ8%を検出器に向かって反 射させることができる。An ideal bandpass interference filter in this case would have a transmittance of 460 nm to 48 nm. The transmittance of wavelengths longer than 500 nm, centering on the 0 nm range, can be ignored. I will do it. Also, other than the conventional wavelength stacking type filter, “square”° bandpass filter or Desired band rejection can also be performed using an induced transmission filter. Furthermore, two Conventional identical filters can also be used in parallel to improve the band rejection ratio. Fi If the outer surface of the router is not coated, approximately 8% of the reflective input light will be reflected back toward the detector. It can be made to shoot.
光源は測定色温度2880°にで作動する石英−ハロゲンランプとした。検出器 は、光起電モードで用いられるUDTピン−10D形のシリコンフォトダイオー ドとした。The light source was a quartz-halogen lamp operating at a measured color temperature of 2880°. Detector is a UDT pin-10D type silicon photodiode used in photovoltaic mode. I did it.
本発明は上述した例にのみ限定されるものではなく要旨を変更しない範囲内で種 々の変更を加えることができる。The present invention is not limited only to the above-mentioned examples, and may be modified without changing the gist thereof. You can make various changes.
国際調を報告 ANNEX To THεINTERNATiONAL 5EARC)l RE POでTONReport on international research ANNEX To THεINTERNAL 5EARC) RE TON at PO
Claims (14)
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
AU937185 | 1985-02-20 | ||
AU9371/85 | 1985-02-20 |
Publications (1)
Publication Number | Publication Date |
---|---|
JPS62501933A true JPS62501933A (en) | 1987-07-30 |
Family
ID=3700123
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP61501506A Pending JPS62501933A (en) | 1985-02-20 | 1986-02-20 | Fluorescence analysis arrangement |
Country Status (3)
Country | Link |
---|---|
EP (1) | EP0245275A1 (en) |
JP (1) | JPS62501933A (en) |
WO (1) | WO1986004988A1 (en) |
Families Citing this family (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
AU587534B2 (en) * | 1986-03-14 | 1989-08-17 | Luminis Pty Limited | Improvements in measurement of fluorescence |
US4804850A (en) * | 1986-03-14 | 1989-02-14 | Luminis Pty. Limited | Measurement of fluorescence |
DE4311543A1 (en) * | 1993-04-07 | 1994-10-13 | Bayerische Motoren Werke Ag | Device for determining the concentration of a test fluid |
GB9606981D0 (en) * | 1996-04-02 | 1996-06-05 | Kodak Ltd | Illumination for scanners |
DE10355164A1 (en) * | 2003-11-26 | 2005-06-23 | Carl Zeiss Jena Gmbh | Process to determine and classify the intensity distribution of a fluorescent microscopic image by comparison of three images |
Family Cites Families (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CH573125A5 (en) * | 1973-11-12 | 1976-02-27 | Balzers Patent Beteilig Ag | |
US4056724A (en) * | 1975-02-27 | 1977-11-01 | International Diagnostic Technology | Fluorometric system, method and test article |
US4022529A (en) * | 1975-12-11 | 1977-05-10 | White John U | Feature extraction system for extracting a predetermined feature from a signal |
GB2052736A (en) * | 1979-06-19 | 1981-01-28 | Sphere Invest | Scanning radiation from moving objects |
US4284412A (en) * | 1979-07-13 | 1981-08-18 | Ortho Diagnostics, Inc. | Method and apparatus for automated identification and enumeration of specified blood cell subclasses |
DE2938056C2 (en) * | 1979-09-20 | 1986-12-11 | Gesellschaft für Strahlen- und Umweltforschung mbH, 8000 München | Device for the fluorometric analysis of samples |
US4336029A (en) * | 1980-08-15 | 1982-06-22 | Ortho Diagnostic Systems Inc. | Method and reagents for quantitative determination of reticulocytes and platelets in whole blood |
EP0114866A1 (en) * | 1982-07-26 | 1984-08-08 | American Hospital Supply Corporation | Improved fluorometer assembly and method |
-
1986
- 1986-02-20 WO PCT/AU1986/000045 patent/WO1986004988A1/en unknown
- 1986-02-20 EP EP86901309A patent/EP0245275A1/en active Pending
- 1986-02-20 JP JP61501506A patent/JPS62501933A/en active Pending
Also Published As
Publication number | Publication date |
---|---|
EP0245275A1 (en) | 1987-11-19 |
WO1986004988A1 (en) | 1986-08-28 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US5091652A (en) | Laser excited confocal microscope fluorescence scanner and method | |
Lakowicz et al. | Instrumentation for fluorescence spectroscopy | |
US5780857A (en) | Apparatus for imaging biochemical samples on substrates | |
CN100480677C (en) | Optical detection apparatus and multi-channel sample analyzer employing the same | |
EP0880690B1 (en) | Fluorescence imaging system compatible with macro and micro scanning objectives | |
JP2653722B2 (en) | Optical read head for immunological test equipment | |
US3971951A (en) | Apparatus for measuring two different fluorescences of a sample | |
CA2157665A1 (en) | An optical arrangement for flow cytometers | |
US20060134775A1 (en) | Systems, illumination subsystems, and methods for increasing fluorescence emitted by a fluorophore | |
JP2002543378A (en) | Apparatus for optically evaluating an object array | |
JPS63500334A (en) | Scattered light measurement device by biological cells for flow cytometer | |
JPH03503454A (en) | Optical reader for immunoassays | |
Maestre et al. | A circular dichroism microspectrophotometer | |
JPS62501933A (en) | Fluorescence analysis arrangement | |
JP2003521670A (en) | Apparatus and method for measuring fluorescence polarization | |
CN205485034U (en) | Coaxial camera of laser | |
JP2016501369A (en) | Optical system, apparatus and method for flow cytometry | |
US4555181A (en) | Apparatus for automatically detecting and evaluating the characteristics of prints | |
JPH01188816A (en) | Spectral type scanning microscope | |
CN220356907U (en) | Multicolor fluorescence excitation optical device | |
WO2022024368A1 (en) | Capillary electrophoresis device | |
RU162184U1 (en) | OPTICAL DEVICE FOR ANALYSIS OF BIOLOGICAL MICROCHIPS | |
SU1256527A1 (en) | Double-beam gas analyzer | |
US2435175A (en) | Flickering beam spectrophotometer for the measurement of bronze | |
RU1805347C (en) | Photometer-fluorimeter-nephelometer |