WO1983001111A1 - Apparatus for the measurement of fluorescence - Google Patents

Apparatus for the measurement of fluorescence Download PDF

Info

Publication number
WO1983001111A1
WO1983001111A1 PCT/FI1982/000036 FI8200036W WO8301111A1 WO 1983001111 A1 WO1983001111 A1 WO 1983001111A1 FI 8200036 W FI8200036 W FI 8200036W WO 8301111 A1 WO8301111 A1 WO 8301111A1
Authority
WO
WIPO (PCT)
Prior art keywords
measurement
monochromator
light
detector
sample
Prior art date
Application number
PCT/FI1982/000036
Other languages
French (fr)
Inventor
Oy Eflab
Original Assignee
Virnes, Martti
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Virnes, Martti filed Critical Virnes, Martti
Publication of WO1983001111A1 publication Critical patent/WO1983001111A1/en

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
    • G01N21/6452—Individual samples arranged in a regular 2D-array, e.g. multiwell plates
    • 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
    • G01N2021/6463—Optics
    • G01N2021/6473—In-line geometry
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2201/00—Features of devices classified in G01N21/00
    • G01N2201/04—Batch operation; multisample devices
    • G01N2201/0446—Multicell plate, sequential

Definitions

  • the subject of the present invention is an apparatus for the measurement of fluorescence out of a liquid sample, the said apparatus comprising a source of light, from which the light of measurement is arranged so as to be passed vertically through optics, a monochromator, and through the sample placed in a pit plate to a measurement monochromator and a detector.
  • the samples have been dosed into small cuvettes, which may also be arranged in the form of a line or plate-
  • the measurement takes place in the longitudinal axial direction of the cuvette so that the cuvette is placed in the apparatus in the space between the source of light required and the detector.
  • a monochromatic excitation light and a detector measuring the fluorescence radi ⁇ ation emitted from the sample are required.
  • a monochromatic excitation light and a detector measuring the fluorescence radi ⁇ ation emitted from the sample are required.
  • a monochromator such as an interference filter or a grid. Since fluorescence radiation is emitted uniformly in all directions, in principle it can be measured from any direction whatsoever. A factor disturbing the measurement is the strong excitation light, wherefore attempts have been made to minimize its effect on the measurement result by means of various arrangements.
  • the commonest mode of measurement in use is the so-called 90° measurement geometry.
  • the directions of the excitation light and of the measure- ment are at an angle of 90 in relation to each other. In this way the access of direct excitation light to the detector is eliminated.
  • the procedure in itself is simple, but it imposes considerable limitations on the geometry of the object to be measured.
  • the sample cuvettes can be treated only one by one or in the line form.
  • It is also possible to measure the fluorescence radiation emitted by the surface by means of a particular arrangement. In this case, the direc ⁇ tions of excitation and measurement form a rather small ( ⁇ 10 to 20°) angle.
  • Such a measurement geometry is usually used when the sample absorbs either the exci- tation radiation or the emitted fluorescence radiation strongly.
  • the apparatus for the measurement of fluore ⁇ scence in accordance with the invention is mainly char ⁇ acterized in that, for each process of measurement, the source of light, the optics, the monochromator, the sample to be measured, the measurement monochromator, and the detector are arranged so that they can be brought onto the same vertical straight line in relation to each other, in this sequence.
  • the invention comes out more closely from the following description and from the attached draw ⁇ ings, wherein
  • Figure 1 is a schematical side view of the apparatus for the measurement of fluorescence and Figure 2 shows a section at A-A in Fig. 1.
  • the source 1 of light and the detector 6 are placed at opposite sides of the sample to be measured (Fig. 1) .
  • the monochromator 5 in front of the detector 6 has been selected so that it is penetrable by the desired measurement wave-length ⁇ i only, but not by the excitation radiation ⁇ o producing the fluorescence.
  • Such a measurement geometry is highly advantageous when such samples are. supposed to be measured as are dosed in a so-called pit plate 4.
  • the most important requirement regarding the source 1 of light is a sufficient light intensity at the wave-length ⁇ o. Suitable sources of light are xenon or mercury gas-discharge lamps. Laser may also be used in some applications. In such a case, no monochromator 3 is required. As a detector 6, it is favourable to use a photomultiplier tube. As monochromators 3 and 5, it is possible to use interference filters or grids. The monochromator 3 and/or the measurement monochromator 5 are preferably exchangeable or removable out of the line of measurement.

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  • 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)

Abstract

An apparatus for the measurement of fluorescence out of a liquid sample, the said apparatus comprising a source (1) of light, from which the light of measurement is arranged to be passed vertically through optics (2), a monochromator (3), and through the sample placed in a pit plate (4) to a measurement monochromator (5) and a detector (6). According to the invention, for each process of measurement, the source (1) of light, the optics (2), the monochromator (3), the sample to be measured, the measurement monochromator (5), and the detector (6) are arranged so that they can be brought onto the same vertical straight line in relation to each other, in this sequence.

Description

Apparatus for the measurement of fluorescence
The subject of the present invention is an apparatus for the measurement of fluorescence out of a liquid sample, the said apparatus comprising a source of light, from which the light of measurement is arranged so as to be passed vertically through optics, a monochromator, and through the sample placed in a pit plate to a measurement monochromator and a detector. The samples have been dosed into small cuvettes, which may also be arranged in the form of a line or plate- The measurement takes place in the longitudinal axial direction of the cuvette so that the cuvette is placed in the apparatus in the space between the source of light required and the detector.
In fluorometers, a monochromatic excitation light and a detector measuring the fluorescence radi¬ ation emitted from the sample are required. As a rule, in front of the detector, there is also some suitable monochromator, such as an interference filter or a grid. Since fluorescence radiation is emitted uniformly in all directions, in principle it can be measured from any direction whatsoever. A factor disturbing the measurement is the strong excitation light, wherefore attempts have been made to minimize its effect on the measurement result by means of various arrangements.
-The commonest mode of measurement in use is the so-called 90° measurement geometry. -.Therein the directions of the excitation light and of the measure- ment are at an angle of 90 in relation to each other. In this way the access of direct excitation light to the detector is eliminated. The procedure in itself is simple, but it imposes considerable limitations on the geometry of the object to be measured. For example, the sample cuvettes can be treated only one by one or in the line form. It is also possible to measure the fluorescence radiation emitted by the surface by means of a particular arrangement. In this case, the direc¬ tions of excitation and measurement form a rather small (~ 10 to 20°) angle. Such a measurement geometry is usually used when the sample absorbs either the exci- tation radiation or the emitted fluorescence radiation strongly.
The apparatus for the measurement of fluore¬ scence in accordance with the invention is mainly char¬ acterized in that, for each process of measurement, the source of light, the optics, the monochromator, the sample to be measured, the measurement monochromator, and the detector are arranged so that they can be brought onto the same vertical straight line in relation to each other, in this sequence. The invention comes out more closely from the following description and from the attached draw¬ ings, wherein
Figure 1 is a schematical side view of the apparatus for the measurement of fluorescence and Figure 2 shows a section at A-A in Fig. 1.
In the measurement arrangement in accordance with the present invention, the source 1 of light and the detector 6 are placed at opposite sides of the sample to be measured (Fig. 1) . The monochromator 5 in front of the detector 6 has been selected so that it is penetrable by the desired measurement wave-length λi only, but not by the excitation radiation λo producing the fluorescence. Such a measurement geometry is highly advantageous when such samples are. supposed to be measured as are dosed in a so-called pit plate 4. The pit plate 4 comprises 8 x 12 = 96 pits, whose volume is max. 300 μl. In the case of a plate, it is impossible to employ the 90° geometry. It is a further advantage of the through measurement that, since the source 1 of light, the sample in the pit plate 4, and the detector 6 can be brought very close to each other, in this way a strong measurement signal is obtained. The same apparatus can also be readily used for absorption measurements by omitting the measurement monochromator 5 placed in front of the detector 6. Also, by means of a suitable arrangement, it is possible to measure the absorption of the sample at the excita¬ tion wave-length λo and at the emission wave-length λi and to calculate an absorption-corrected fluorescence signal out of them.
The most important requirement regarding the source 1 of light is a sufficient light intensity at the wave-length λo. Suitable sources of light are xenon or mercury gas-discharge lamps. Laser may also be used in some applications. In such a case, no monochromator 3 is required. As a detector 6, it is favourable to use a photomultiplier tube. As monochromators 3 and 5, it is possible to use interference filters or grids. The monochromator 3 and/or the measurement monochromator 5 are preferably exchangeable or removable out of the line of measurement.

Claims

WHAT IS CLAIMED IS:
1. An apparatus for the measurement of fluorescence out of a liquid sample, the said apparatus comprising a source (1) of light, from which the light of measurement is arranged to be passed vertically through optics (2) , a monochromator (3) , and through the sample placed in a pit plate (4) to a measurement monochromator (5) and a detector (6), c h a r a c - - t e r i z e d in that, for each process of measurement, the source (1) of light, the optics (2) , the monochro¬ mator (3) , the sample to be measured, the measurement monochromator (5) , and the detector (6) are arranged so that they can be brought onto the same vertical straight line in relation to each other, in this sequence.
2. An. apparatus as claimed in claim 1 , c h a r a c t e r i z e d in that the monochromator (3) and/or the measurement monochromator (5) can be exchanged or removed out of the line of measurement.
PCT/FI1982/000036 1981-09-21 1982-09-20 Apparatus for the measurement of fluorescence WO1983001111A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
FI812933A FI812933L (en) 1981-09-21 1981-09-21 ANORDINATION FOR MAINTENANCE OF FLUORESCENS
FI812933810921 1981-09-21

Publications (1)

Publication Number Publication Date
WO1983001111A1 true WO1983001111A1 (en) 1983-03-31

Family

ID=8514714

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/FI1982/000036 WO1983001111A1 (en) 1981-09-21 1982-09-20 Apparatus for the measurement of fluorescence

Country Status (4)

Country Link
EP (1) EP0088781A1 (en)
JP (1) JPS58501735A (en)
FI (1) FI812933L (en)
WO (1) WO1983001111A1 (en)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0108524A1 (en) * 1982-10-12 1984-05-16 Dynatech Laboratories, Incorporated Fluorometer
EP0135303A2 (en) * 1983-08-24 1985-03-27 Labsystems Oy Set of cuvettes
US4810096A (en) * 1986-05-09 1989-03-07 Cambridge Life Sciences, Plc Plate reader

Citations (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
SE364367B (en) * 1967-05-12 1974-02-18 Rec Scient Centre Nat
DE2451769A1 (en) * 1973-11-14 1975-05-15 Osmo Antero Suovaniemi METHOD AND DEVICE FOR AUTOMATIC READING AND DETERMINATION OF REACTION RESULTS
DE2104393B2 (en) * 1971-01-30 1976-01-02 Original Hanau Quarzlampen Gmbh, 6450 Hanau Fluorescence analyser with mercury discharge lamp - designed for investigating fluorescent radiation from organic and inorganic substances
SE383207B (en) * 1972-08-22 1976-03-01 Atomic Energy Commission COMBINED DYNAMIC PHOTOMETER AND FLUORMETER OF MULTIPLE STATION TYPE.
FR2408136A1 (en) * 1977-11-04 1979-06-01 Colin Bruno Determining effects of different analysis reactive agents - utilises focussing of luminous beam for measurement of optical density
GB2014300A (en) * 1977-07-29 1979-08-22 Opto Electronic Displays Ltd Opacity measurement apparatus and method
DE2818000A1 (en) * 1978-04-25 1979-11-08 Gunnar Oosterloo Comparative fluorescence spectroscopy device for chemistry teaching - contains ultraviolet source, sample slide-in unit and fluorescent foil
FR2430610A1 (en) * 1978-07-07 1980-02-01 Pasteur Institut METHOD AND DEVICE FOR DETERMINING THE SENSITIVITY OF BACTERIA AND YEAST TO ANTIBIOTIC PRODUCTS
EP0012698A1 (en) * 1978-12-18 1980-06-25 Merck & Co. Inc. Automated apparatus and method for assay plate reading
WO1982000358A1 (en) * 1980-07-24 1982-02-04 Oy Labsystems Method of measurement and a cuvette
WO1982000359A1 (en) * 1980-07-24 1982-02-04 Oy Labsystems Set of cuvettes
EP0046430A1 (en) * 1980-08-14 1982-02-24 Commissariat à l'Energie Atomique Method for real-time detection and quantification of agglutinates

Patent Citations (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
SE364367B (en) * 1967-05-12 1974-02-18 Rec Scient Centre Nat
DE2104393B2 (en) * 1971-01-30 1976-01-02 Original Hanau Quarzlampen Gmbh, 6450 Hanau Fluorescence analyser with mercury discharge lamp - designed for investigating fluorescent radiation from organic and inorganic substances
SE383207B (en) * 1972-08-22 1976-03-01 Atomic Energy Commission COMBINED DYNAMIC PHOTOMETER AND FLUORMETER OF MULTIPLE STATION TYPE.
DE2451769A1 (en) * 1973-11-14 1975-05-15 Osmo Antero Suovaniemi METHOD AND DEVICE FOR AUTOMATIC READING AND DETERMINATION OF REACTION RESULTS
GB2014300A (en) * 1977-07-29 1979-08-22 Opto Electronic Displays Ltd Opacity measurement apparatus and method
FR2408136A1 (en) * 1977-11-04 1979-06-01 Colin Bruno Determining effects of different analysis reactive agents - utilises focussing of luminous beam for measurement of optical density
DE2818000A1 (en) * 1978-04-25 1979-11-08 Gunnar Oosterloo Comparative fluorescence spectroscopy device for chemistry teaching - contains ultraviolet source, sample slide-in unit and fluorescent foil
FR2430610A1 (en) * 1978-07-07 1980-02-01 Pasteur Institut METHOD AND DEVICE FOR DETERMINING THE SENSITIVITY OF BACTERIA AND YEAST TO ANTIBIOTIC PRODUCTS
EP0012698A1 (en) * 1978-12-18 1980-06-25 Merck & Co. Inc. Automated apparatus and method for assay plate reading
WO1982000358A1 (en) * 1980-07-24 1982-02-04 Oy Labsystems Method of measurement and a cuvette
WO1982000359A1 (en) * 1980-07-24 1982-02-04 Oy Labsystems Set of cuvettes
EP0046430A1 (en) * 1980-08-14 1982-02-24 Commissariat à l'Energie Atomique Method for real-time detection and quantification of agglutinates

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0108524A1 (en) * 1982-10-12 1984-05-16 Dynatech Laboratories, Incorporated Fluorometer
EP0135303A2 (en) * 1983-08-24 1985-03-27 Labsystems Oy Set of cuvettes
EP0135303B1 (en) * 1983-08-24 1988-11-02 Labsystems Oy Set of cuvettes
US4810096A (en) * 1986-05-09 1989-03-07 Cambridge Life Sciences, Plc Plate reader

Also Published As

Publication number Publication date
JPS58501735A (en) 1983-10-13
FI812933L (en) 1983-03-22
EP0088781A1 (en) 1983-09-21

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