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DE20104446U1 - Fluorescence standard - Google Patents

Fluorescence standard

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Publication number
DE20104446U1
DE20104446U1 DE20104446U DE20104446U DE20104446U1 DE 20104446 U1 DE20104446 U1 DE 20104446U1 DE 20104446 U DE20104446 U DE 20104446U DE 20104446 U DE20104446 U DE 20104446U DE 20104446 U1 DE20104446 U1 DE 20104446U1
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DE
Germany
Prior art keywords
carrier
fluorescence standard
fluorescence
fluorescent substance
weight
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.)
Expired - Lifetime
Application number
DE20104446U
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German (de)
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Pes Gesellschaft fur Medizinische Diagnose-Sy De
Original Assignee
Pe Diagnostik GmbH
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 Pe Diagnostik GmbH filed Critical Pe Diagnostik GmbH
Priority to DE20104446U priority Critical patent/DE20104446U1/en
Publication of DE20104446U1 publication Critical patent/DE20104446U1/en
Anticipated expiration legal-status Critical
Expired - Lifetime legal-status Critical Current

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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
    • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
    • G01N21/27—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands using photo-electric detection ; circuits for computing concentration
    • G01N21/274—Calibration, base line adjustment, drift correction
    • G01N21/278—Constitution of standards
    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09D—COATING COMPOSITIONS, e.g. PAINTS, VARNISHES OR LACQUERS; FILLING PASTES; CHEMICAL PAINT OR INK REMOVERS; INKS; CORRECTING FLUIDS; WOODSTAINS; PASTES OR SOLIDS FOR COLOURING OR PRINTING; USE OF MATERIALS THEREFOR
    • C09D5/00—Coating compositions, e.g. paints, varnishes or lacquers, characterised by their physical nature or the effects produced; Filling pastes
    • C09D5/22—Luminous paints
    • 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
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/52—Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements
    • 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
    • G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
    • G01N2021/6432—Quenching
    • 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
    • G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
    • G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
    • G01N2021/7769—Measurement method of reaction-produced change in sensor
    • G01N2021/7786—Fluorescence

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • Hematology (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Urology & Nephrology (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • Materials Engineering (AREA)
  • Biotechnology (AREA)
  • Cell Biology (AREA)
  • Wood Science & Technology (AREA)
  • Organic Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Mathematical Physics (AREA)
  • Theoretical Computer Science (AREA)
  • Spectroscopy & Molecular Physics (AREA)
  • Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)

Description

FluoreszenzstandardFluorescence standard

Die Erfindung betrifft einen Fluoreszenzstandard, bestehend aus einer Referenzfläche genau bekannter Fluoreszenz einer fluoreszierenden Substanz auf einem Träger.The invention relates to a fluorescence standard consisting of a reference surface of precisely known fluorescence of a fluorescent substance on a carrier.

Ein Fluoreszenzstandard ist eine Referenzfläche, die eine sehr genau bekannte Fluoreszenz liefert. Diese Fläche wird durch Aufbringen einer fluoreszierenden Substanz auf einen Träger hergestellt. Der Träger kann auf der Messfläche eines Sensors als auch auf einer davon räumlich getrennten Stelle positioniert sein. Als Sensor sind auch solche Konstruktionen zu bezeichnen, die keine eigentliche Sensorfunktionen besitzen, sondern nur für Kalibrierzwecke eingesetzt werden. Dazu ist der Fluoreszenzstandard in den Messraum des zu kalibrierenden bzw. abzugleichenden Messgerätes eingebracht. Die Kalibration von Fluoreszenzmessgeräten ist an langzeitstabile Normale gekoppelt, die kaum verfügbar und technisch für spezielle geometrische Formen eines Trägers nur schwer herstellbar sind.A fluorescence standard is a reference surface that provides a very precisely known fluorescence. This surface is produced by applying a fluorescent substance to a carrier. The carrier can be positioned on the measuring surface of a sensor or at a spatially separate location. Constructions that do not have any actual sensor functions but are only used for calibration purposes can also be referred to as sensors. For this purpose, the fluorescence standard is introduced into the measuring space of the measuring device to be calibrated or adjusted. The calibration of fluorescence measuring devices is linked to long-term stable standards that are hardly available and are technically difficult to produce for special geometric shapes of a carrier.

Aufgabe der Erfindung ist es, einen Fluoreszenzstandard, bestehend aus einer Referenzfläche genau bekannter Fluoreszenz einer fluoreszierenden Substanz auf einem Träger zu entwickeln, wobei der Fluoreszenzstandard leicht herstellbar und für alle geometrische Formen der Träger verwendbar ist.The object of the invention is to develop a fluorescence standard consisting of a reference surface of precisely known fluorescence of a fluorescent substance on a carrier, whereby the fluorescence standard is easy to produce and can be used for all geometric shapes of the carrier.

a a · ··· ■·· e*a· · aa a · ··· ■·· e*a· · a

Φ &bgr; &bgr; &bgr;Φ β β β

Erfindungsgemäß wird die Aufgabe dadurch gelöst, dass die fluoreszierende Substanz in flüssiger und lackähnlicher Form auf einen Träger jeder geometrischen Form aufbringbar und darauf aushärtbar ist, und die fluoreszierende Substanz ausAccording to the invention, the object is achieved in that the fluorescent substance can be applied in liquid and lacquer-like form to a carrier of any geometric shape and can be cured thereon, and the fluorescent substance consists of

- einer wässrigen Kunststoffdispersion aus 0,5 bis 50 Gew.-% Styrol-Acrylsäureester-Copolymerisat und 50 bis 99,5 Gew.-% Wasser und- an aqueous plastic dispersion of 0.5 to 50 wt.% styrene-acrylic acid ester copolymer and 50 to 99.5 wt.% water and

- einer Fluorophorlösung mit 0,00001 bis 0,001 Gew.-% Farbstoff (C46H53N4NaO14S2 oder C40H40N6O6S), der in 99,999 bis 99,99999 Gew.-% Wasser gelöst ist, besteht, die im Verhältnis von 90 bis 99,9 Vol.-% wässrige Kunststoffdispersion und 0,1 bis 10 Vol.-% Fluorophorlösung gemischt sind.- a fluorophore solution containing 0.00001 to 0.001% by weight of dye (C 46 H 53 N 4 NaO 14 S 2 or C 40 H 40 N 6 O 6 S) dissolved in 99.999 to 99.99999% by weight of water, mixed in a ratio of 90 to 99.9% by volume of aqueous plastic dispersion and 0.1 to 10% by volume of fluorophore solution.

Zweckmäßig ist es, dass der Träger ein Biosensor ist, bei dem der Fluoreszenzstandard als Streifen oder Punktreihe ausgebildet ist. In einer weiteren Ausführungsform der Erfindung ist der Träger ein Prüfkörper zu Kalibrationszwecken, bei dem der Fluoreszenzstandard auf die gesamte Messfläche des Prüfkörpers oder Teilen davon aufgetragen ist.It is expedient for the carrier to be a biosensor in which the fluorescence standard is designed as a strip or row of dots. In a further embodiment of the invention, the carrier is a test body for calibration purposes in which the fluorescence standard is applied to the entire measuring surface of the test body or parts thereof.

Nachfolgend wird die Erfindung anhand eines Ausführungsbeispieles näher erläutert. Es wird eine wässrige Kunststoffdispersion aus 10 Gew.-% Styrol-Acrylsäureester-Copolymerisat mit der Typenbezeichnung „VINNAPAS SAF 54" der Fa. Wacker Polymer Systems GmbH & Co. KG und 90 Gew.-% Wasser hergestellt. Nachfolgend wird eine Fluorophorlösung hergestellt, indem 0,001 Gew.-% Farbstoff (C46H53N4NaOi4S2 oder C40H40N6O6S) in 99,999 Gew.-% Wasser gelöst ist. Die wässrige Kunststoffdispersion und die Fluorophorlösung werden in einem Verhältnis von 90 Vol.-% wässrige Kunststoffdispersion und Vol.-% Fluorophorlösung zu einer fluoreszierenden Substanz gemischt. Die fluoreszierende Substanz wird in flüssiger und lackähnlicher Form als Referenzfläche genau bekannter Fluoreszenz auf einem Träger zum Aushärten aufgebracht. Die Beschichtung dieses Trägers als Fluoreszenzstandard kannThe invention is explained in more detail below using an example. An aqueous plastic dispersion is prepared from 10% by weight of styrene-acrylic acid ester copolymer with the type designation "VINNAPAS SAF 54" from Wacker Polymer Systems GmbH & Co. KG and 90% by weight of water. A fluorophore solution is then prepared by dissolving 0.001% by weight of dye (C 46 H 53 N 4 NaOi 4 S 2 or C 40 H 40 N 6 O 6 S) in 99.999% by weight of water. The aqueous plastic dispersion and the fluorophore solution are mixed in a ratio of 90% by volume of aqueous plastic dispersion and 90% by volume of fluorophore solution to form a fluorescent substance. The fluorescent substance is applied in liquid and paint-like form to a carrier for curing as a reference surface of precisely known fluorescence. The coating of this carrier as a fluorescence standard can

■ &agr; &lgr; &lgr;, m „. _■ &agr;&lgr; λ, m ". _

entweder lokal, beispielsweise auf einem Biosensor als Streifen, oder auch homogen, beispielsweise über die gesamte Messfläche eines ortsauflösenden Fluorimeters, erfolgen. Die aufgebrachte fluoreszierende Substanz trocknet an Luft zu einem wasserfesten Film aus und ist somit auf jede beliebige geometrische Form aufbringbar. Die spezifische Fluoreszenzmarkierung wird durch gezieltes Einbringen von Fluorophor (Farbstoff) erreicht. Bei homogen aufgetragener flächiger Verteilung lassen sich ortsabhängige Empfindlichkeitskurven von ortsauflösend messenden Fluorimetern gut bestimmen und damit kalibrieren. Die ortsbezogene Homogenität der ausgehärteten aufgebrachten Substanz auf der Messfläche kann unabhängig von einem Messsystem mit bekannter homogener Ortskennlinie bestimmt werden. Dazu fertigt man eine Serie von beispielsweise 10 Referenzflächen an und vermisst diese nacheinander mit demselben Messgerät. Anschließend wird für jede Ortsposition die Varianz von Referenzfläche zu Referenzfläche berechnet. Diese Varianz beinhaltet neben möglichen Inhomogenitäten des Fluoreszenzstandards auch den Gerätefehler des verwendeten Messsystems. Dieser wird dadurch bestimmt, dass man zu der Serie von Referenzflächen eine einzige auswählt und z. B. 10 mal in demselben Messgerät vermisst. Auch hier berechnet man wieder die Varianz für jede Ortsposition von Messung zu Messung und erhält damit den reinen Messgerätefehler. Durch den Vergleich der beiden Varianzkennlinien kann die Homogenität des Fluoreszenzstandards beurteilt werden, ohne die Empfindlichkeitskennlinie des Messgerätes zu kennen. Ist die Homogenität des Fluoreszenzstandards deutlich besser als der reine Gerätefehler des Messsystems, so entspricht die Messkurve der hergestellten Referenzflächen der ortsabhängigen Empfindlichkeitskennlinie des Messgerätes.either locally, for example on a biosensor as a strip, or homogeneously, for example over the entire measuring surface of a spatially resolving fluorimeter. The applied fluorescent substance dries in air to form a waterproof film and can therefore be applied to any geometric shape. The specific fluorescence marking is achieved by the targeted introduction of fluorophore (dye). With a homogeneous application over a large area, location-dependent sensitivity curves of spatially resolving fluorimeters can be easily determined and thus calibrated. The location-related homogeneity of the hardened substance applied to the measuring surface can be determined independently of a measuring system with a known homogeneous location characteristic curve. To do this, a series of, for example, 10 reference surfaces is prepared and these are measured one after the other with the same measuring device. The variance from reference surface to reference surface is then calculated for each location. In addition to possible inhomogeneities in the fluorescence standard, this variance also includes the device error of the measuring system used. This is determined by selecting a single reference surface from the series and measuring it 10 times in the same measuring device, for example. Here, too, the variance for each position is calculated from measurement to measurement, thus obtaining the pure measuring device error. By comparing the two variance curves, the homogeneity of the fluorescence standard can be assessed without knowing the sensitivity curve of the measuring device. If the homogeneity of the fluorescence standard is significantly better than the pure device error of the measuring system, the measurement curve of the reference surfaces produced corresponds to the location-dependent sensitivity curve of the measuring device.

Claims (3)

1. Fluoreszenzstandard, bestehend aus einer Referenzfläche genau bekannter Fluoreszenz einer fluoreszierenden Substanz auf einem Träger, dadurch gekennzeichnet, dass die fluoreszierende Substanz in flüssiger und lackähnlicher Form auf einen Träger jeder geometrischen Form aufbringbar und darauf aushärtbar ist, und die fluoreszierende Substanz aus - einer wässrigen Kunststoffdispersion aus 0,5 bis 50 Gew.-% Styrol- Acrylsäureester-Copolymerisat und 50 bis 99,5 Gew.-% Wasser und - einer Fluorophorlösung mit 0,00001 bis 0,001 Gew.-% Farbstoff (C46H53N4NaO14S2 oder C40H40N6O6S), der in 99,999 bis 99,99999 Gew.-% Wasser gelöst ist, besteht, die im Verhältnis von 90 bis 99,9 Vol.-% wässrige Kunststoffdispersion und 0,1 bis 10 Vol.-% Fluorophorlösung gemischt sind. 1. Fluorescence standard consisting of a reference surface of precisely known fluorescence of a fluorescent substance on a carrier, characterized in that the fluorescent substance can be applied in liquid and lacquer-like form to a carrier of any geometric shape and can be cured thereon, and the fluorescent substance consists of - an aqueous plastic dispersion of 0.5 to 50 wt.% styrene-acrylic acid ester copolymer and 50 to 99.5 wt.% water and - a fluorophore solution containing 0.00001 to 0.001% by weight of dye (C 46 H 53 N 4 NaO 14 S 2 or C 40 H 40 N 6 O 6 S) dissolved in 99.999 to 99.99999% by weight of water, mixed in a ratio of 90 to 99.9% by volume of aqueous plastic dispersion and 0.1 to 10% by volume of fluorophore solution. 2. Fluoreszenzstandard nach Anspruch 1, dadurch gekennzeichnet, dass der Träger ein Biosensor ist, bei dem der Fluoreszenzstandard als Streifen oder Punktreihe ausgebildet ist. 2. Fluorescence standard according to claim 1, characterized in that the carrier is a biosensor in which the fluorescence standard is designed as a strip or row of dots. 3. Fluoreszenzstandard nach Anspruch 1, dadurch gekennzeichnet, dass der Träger ein Prüfkörper zu Kalibrationszwecken ist, bei dem der Fluoreszenzstandard auf die gesamte Messfläche des Prüfkörpers oder Teilen davon aufgetragen ist. 3. Fluorescence standard according to claim 1, characterized in that the carrier is a test body for calibration purposes, in which the fluorescence standard is applied to the entire measuring surface of the test body or parts thereof.
DE20104446U 2001-03-13 2001-03-13 Fluorescence standard Expired - Lifetime DE20104446U1 (en)

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
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
DE102004047593A1 (en) * 2004-09-30 2006-04-13 Carl Zeiss Jena Gmbh Reference body for fluorescence measurements and method of making the same
DE102007038754A1 (en) * 2007-08-16 2009-02-19 Giesecke & Devrient Gmbh Device for the calibration of sensors
EP3926022A1 (en) * 2020-06-18 2021-12-22 IMT Masken und Teilungen AG Calibration target

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE3148912A1 (en) 1981-12-10 1983-06-23 Wolfgang Dr. 6301 Pohlheim Oberheim Mercury reference standard
DE4300723C2 (en) 1993-01-14 1995-10-05 Boehringer Mannheim Gmbh Device for measuring the fluorescence of a sample, in particular for the medical-analytical determination of the concentration of a component of a body fluid
DE19808591A1 (en) 1998-02-28 1999-09-16 Univ Leipzig Standardized flow-through cytometric whole blood assay
DE19847370A1 (en) 1998-10-14 2000-04-20 Heinz Langhals Use of perylene tetracarbobisimide dyes, tetraesters and tetracarboxylates are used as standard substances in fluorimetry and of bisanhydride as starting material
DE19848555A1 (en) 1998-10-21 2000-04-27 Heinz Langhals New perylene-3,4,9,10-tetracarboxylic acid bis-imide derivatives and analogs, used as dyes, especially fluorescent dyes, e.g. for bulk dyeing plastics, vat dyeing natural fibers or staining DNA
EP1031828A1 (en) 1999-02-25 2000-08-30 C.S.E.M. Centre Suisse D'electronique Et De Microtechnique Sa Integrated-optical sensor and method for integrated-optically sensing a substance

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE3148912A1 (en) 1981-12-10 1983-06-23 Wolfgang Dr. 6301 Pohlheim Oberheim Mercury reference standard
DE4300723C2 (en) 1993-01-14 1995-10-05 Boehringer Mannheim Gmbh Device for measuring the fluorescence of a sample, in particular for the medical-analytical determination of the concentration of a component of a body fluid
DE19808591A1 (en) 1998-02-28 1999-09-16 Univ Leipzig Standardized flow-through cytometric whole blood assay
DE19847370A1 (en) 1998-10-14 2000-04-20 Heinz Langhals Use of perylene tetracarbobisimide dyes, tetraesters and tetracarboxylates are used as standard substances in fluorimetry and of bisanhydride as starting material
DE19848555A1 (en) 1998-10-21 2000-04-27 Heinz Langhals New perylene-3,4,9,10-tetracarboxylic acid bis-imide derivatives and analogs, used as dyes, especially fluorescent dyes, e.g. for bulk dyeing plastics, vat dyeing natural fibers or staining DNA
EP1031828A1 (en) 1999-02-25 2000-08-30 C.S.E.M. Centre Suisse D'electronique Et De Microtechnique Sa Integrated-optical sensor and method for integrated-optically sensing a substance

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
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
DE102004047593A1 (en) * 2004-09-30 2006-04-13 Carl Zeiss Jena Gmbh Reference body for fluorescence measurements and method of making the same
DE102007038754A1 (en) * 2007-08-16 2009-02-19 Giesecke & Devrient Gmbh Device for the calibration of sensors
EP3926022A1 (en) * 2020-06-18 2021-12-22 IMT Masken und Teilungen AG Calibration target
US11543350B2 (en) 2020-06-18 2023-01-03 IMT Masken und Teilungen AG Calibration target

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