EP2292329A9 - Substrat polymère doté d'une structure fluorescente, son procédé de fabrication et son utilisation - Google Patents

Substrat polymère doté d'une structure fluorescente, son procédé de fabrication et son utilisation Download PDF

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Publication number
EP2292329A9
EP2292329A9 EP09011507A EP09011507A EP2292329A9 EP 2292329 A9 EP2292329 A9 EP 2292329A9 EP 09011507 A EP09011507 A EP 09011507A EP 09011507 A EP09011507 A EP 09011507A EP 2292329 A9 EP2292329 A9 EP 2292329A9
Authority
EP
European Patent Office
Prior art keywords
polymer substrate
fluorescent
sample chamber
fluorescence
cover plate
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.)
Granted
Application number
EP09011507A
Other languages
German (de)
English (en)
Other versions
EP2292329B1 (fr
EP2292329A1 (fr
Inventor
Thomas Fischer
Joachim Stumpe
Valentin Kahl
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.)
Ibidi GmbH
Fraunhofer Gesellschaft zur Foerderung der Angewandten Forschung eV
Original Assignee
Ibidi GmbH
Fraunhofer Gesellschaft zur Foerderung der Angewandten Forschung eV
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 Ibidi GmbH, Fraunhofer Gesellschaft zur Foerderung der Angewandten Forschung eV filed Critical Ibidi GmbH
Priority to EP09011507.2A priority Critical patent/EP2292329B1/fr
Priority to US12/876,881 priority patent/US9597688B2/en
Publication of EP2292329A1 publication Critical patent/EP2292329A1/fr
Publication of EP2292329A9 publication Critical patent/EP2292329A9/fr
Application granted granted Critical
Publication of EP2292329B1 publication Critical patent/EP2292329B1/fr
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Classifications

    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/54Labware with identification means
    • B01L3/545Labware with identification means for laboratory containers

Definitions

  • the invention relates to polymer substrates provided with fluorescence features in which fluorescence characteristics, ie fluorescent structures, are produced photochemically by UV irradiation.
  • fluorescence characteristics ie fluorescent structures
  • suitable fluorophors can be produced by suitable UV irradiation, which exhibit marked and detectable emission upon excitation with light of suitable wavelength. If such irradiation is carried out in a structured manner, emission patterns can be generated in this way in polymer substrates, which can be used, for example, as a recovery grid in fluorescence microscopy.
  • Another field of application relates to product authentication, which is made possible by the polymer substrates provided with fluorescence features according to the invention.
  • sample chambers In the field of culturing cells, a wide variety of sample chambers are used. These are mostly polymer-based and range from the cell culture bottle to slides and ⁇ -slides from ibidi to multiwell plates.
  • sample chambers can have contiguous areas on which cells grow from 1 mm 2 to 100 cm 2 . Since cells typically have a diameter of 1 .mu.m to 30 .mu.m, finding individual cells in large-area chambers without recovery structures is almost impossible.
  • the EP 2 008 715 A1 also describes a recovery grid, which is formed as part of the plastic body or is introduced into the plastic body.
  • the systems described here are based on non-fluorescent recovery gratings.
  • Another area of the prior art underlying the present invention relates to the authentication of products, especially consumer products.
  • fluorescence films can be structured by bleaching existing autofluorescence.
  • a polymer substrate having a fluorescent structure is provided as an integral constituent, wherein a fluorescent structure is generated photochemically in regions in the polymer substrate.
  • the preferred polymer substrates are low intrinsic fluorescence polymers, i. with an intrinsic fluorescence on the order of a cover glass in the excitation and emission range from 200 nm to 1000 nm. These include in particular COC, COP, PMMA, aliphatic polyesters and polyurethanes, and also polyethers. Particularly good polymers are suitable without UV stabilizers. For optical microscopy, in particular polymers with a refractive index between 1.4 and 1.6, in particular with 1.51 and / or a Abbe number over 50 and / or with a low birefringence are suitable.
  • a fluorescent structure is understood to mean structures up to the optical resolution limit. Typically, however, structures with a lateral resolution of several ⁇ m are used.
  • the fluorescent structure should have at least twice as high intensity as the non-fluorescent structure.
  • the structure should have such a high intensity that it can be easily recognized even at exposure times of less than one second with commercial fluorescence microscopes. This is already the case with a factor of 10.
  • the fluorophores which form the fluorescent structure are firmly bound in the polymer substrate and form a unit with it.
  • the fluorescent structures are thus an integral part of the carrier.
  • the modified polymer is characterized by the fact that both the fluorescent regions and the non-fluorescent regions consist of the identical original material. Therefore, the fluorophores also can not diffuse or sweat out.
  • a structuring of the polymer substrate with fluorescent regions is produced.
  • a mask or a locally positionable radiation source is preferably used. If the UV irradiation is carried out in a structured manner, only the areas that have been irradiated will fluoresce significantly. In the non-irradiated areas, however, no substantial increase in the base emission is achieved. In this way, patterns can be generated which become visible upon excitation with light of suitable wavelength.
  • Such a photochemical structuring produces patterned fluorescent areas that are long-term stable and stable to environmental influences. Furthermore, they are characterized by the fact that they do not lose their edge sharpness by diffusion or similar processes. In addition, the features thus produced can not be removed without trace.
  • the fluorescent structure in the polymer substrate consists of several elements in the form of bars, lines, characters, figures, interference patterns, or combinations thereof.
  • fluorescent structures can be generated with only limited by the substrate size limited overall extent of the pattern.
  • the lateral resolution of the elements of the fluorescent structure is limited only by the method of introduction and the wavelength of the light.
  • the individual structural elements of the fluorescence pattern typically have a lateral extent of less than 100 ⁇ m, preferably less than 10 ⁇ m.
  • a method is also provided for producing the polymer substrate described above, in which a polymer substrate is at least partially subjected to irradiation in the wavelength range below 300 nm to produce fluorescent structures.
  • the preparation of the fluorescent structures by UV irradiation using masks by imaging or contact exposure takes place.
  • plates which have an exemplary variation of regions that are either transparent or non-transparent to the irradiation light used.
  • both radiation sources with coherent (laser direct writing) and focused beam (sample positioning) can be used for structuring.
  • Radiation sources used are preferably radiation sources with emission wavelengths or emission wavelength ranges in the range below 300 nm. These include, in particular, deuterium lamps, excimer lamps (Xe), excimer lasers (F 2 , ArF, KrF) or solid-state lasers (Nd: YVO 4 / YLF).
  • the polymer substrate used is preferably a polymer film.
  • the UV-generated generation of the fluorophores can be monitored in the absorption spectrum of the polymer substrate by an increase in the extinction in the UV range. However, these changes do not significantly affect the optical transmissivity in the visible region of the light to the human eye.
  • the polymer substrates can be provided without damage on the front or back with fluorescent features. Their production requires no exclusion of atmospheric oxygen.
  • the fluorescent features are further characterized by the fact that they are generated directly in materials that are used anyway for products from different areas. There are no additional coatings, imprints or other application fluorophores, fluorescent labels or their precursors. The fact that the features are produced directly in the material used, makes it much easier and cheaper to manufacture this compared to other methods. In addition, it is a fully optical process that does not require any wet-chemical development processes.
  • Such emission patterns can be used to provide substrates for fluorescence microscopy with a recovery grid.
  • the recovery grids are designed as a grid, which is also provided with numbers or letters. These recovery grids facilitate the retrieval of certain sample areas by being precisely charac- terised by the grid grid.
  • the recovery grids do not significantly affect biological processes and are sufficiently stable to perform longer observations on biological and other samples with greater reliability and reproducibility.
  • the described, photochemically generated characteristics are very well suited for typical excitation conditions in fluorescence microscopy; they are visible in all fluorescence channels (blue, green, yellow and red).
  • the lateral resolution in such produced features is sufficiently high, e.g. to allow a mapped observation of cells of different types.
  • Chambers in which such gratings can be introduced are in the DE 100 04 135 , the DE 101 05 711 , the EP 02 777 215 , the EP 05 041 563 , the EP 06 015 167 , the EP 07 012 400 and the EP 09 006 487 described.
  • the upper part has at least one recess.
  • By connecting to the lower part of a reservoir is formed.
  • the recess of the upper part of the reservoir is then designed as a closed channel / tube or as an open-topped container.
  • the bottom formed by the base may be a foil or a coated glass carrier.
  • the bottom has a preferred thickness of 50 .mu.m to 250 .mu.m and / or an Abbe number greater than 50 and / or a refractive index between 1.2 and 1.8, in particular between 1.45 and 1.55. These properties are particularly well suited for (high-resolution) microscopy.
  • the bottom or film can be irradiated both from the side on which the cells are to be cultivated later and from the corresponding opposite side in order to produce fluorescent structures.
  • microfluidic analysis chambers can be provided with corresponding features.
  • coated glass supports can also be used.
  • Glass cover glasses are coated with a COP or COC layer to then introduce into this layer via the method described fluorescent grating.
  • the layer may e.g. be applied by spincoating etc.
  • labels, labels and products made of polymers can be provided with a feature by the structured UV irradiation, which becomes clearly visible only by excitation with light of suitable wavelength and intensity.
  • the counterfeiting security of the product increases and a non-erasable product individualization can be carried out, for example via barcodes.
  • This can be realized by a laser marking system, wherein the feature can also be read electronically.
  • Fig. 1 shows an emission spectrum of a polymer substrate according to the invention before and after the generation of fluorescence regions.
  • the irradiation for the generation of the fluorescence regions took place here with an ArF excimer laser.
  • the irradiation time was 10 seconds.
  • a polymer substrate according to the invention which has number patterns which were produced with the aid of a mask.
  • This is a fluorescence micrograph under excitation at a wavelength of 365 nm and 200x magnification.
  • a mask pattern chrome on silica glass
  • the mask rests with the chrome side. It is irradiated with a 30W deuterium lamp for 3 hours. Then the mask is removed and the emission pattern can be visualized by excitation at 365 nm.
  • a mask pattern chrome on silica glass
  • the mask rests with the chrome side. It is irradiated with an ArF excimer laser (193 nm) for 10 seconds. The mask is then removed and the emission pattern can be visualized by excitation at 365 nm, 436 nm or 515 nm.

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  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Clinical Laboratory Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
EP09011507.2A 2009-09-08 2009-09-08 Substrat polymère doté d'une structure fluorescente, son procédé de fabrication et son utilisation Active EP2292329B1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
EP09011507.2A EP2292329B1 (fr) 2009-09-08 2009-09-08 Substrat polymère doté d'une structure fluorescente, son procédé de fabrication et son utilisation
US12/876,881 US9597688B2 (en) 2009-09-08 2010-09-07 Polymer substrate with fluorescent structure, method for the production thereof and the use thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
EP09011507.2A EP2292329B1 (fr) 2009-09-08 2009-09-08 Substrat polymère doté d'une structure fluorescente, son procédé de fabrication et son utilisation

Publications (3)

Publication Number Publication Date
EP2292329A1 EP2292329A1 (fr) 2011-03-09
EP2292329A9 true EP2292329A9 (fr) 2011-05-04
EP2292329B1 EP2292329B1 (fr) 2014-11-12

Family

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Application Number Title Priority Date Filing Date
EP09011507.2A Active EP2292329B1 (fr) 2009-09-08 2009-09-08 Substrat polymère doté d'une structure fluorescente, son procédé de fabrication et son utilisation

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US (1) US9597688B2 (fr)
EP (1) EP2292329B1 (fr)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11977028B2 (en) * 2020-04-15 2024-05-07 Northwestern University Mechanical-bond-induced exciplex fluorescence
LU101796B1 (de) * 2020-05-14 2021-11-15 Phoenix Contact Gmbh & Co Laserbeschriftbares Kennzeichnungsschild

Family Cites Families (19)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE4241663C2 (de) * 1992-12-04 1997-05-28 Borus Spezialverfahren Verfahren zur Kennzeichnung und Erkennung eines Gegenstands
JP3068483B2 (ja) * 1996-02-08 2000-07-24 株式会社東芝 パターン読み出し方法、および光学的信号読み取り装置
FR2755902B1 (fr) * 1996-11-20 1999-02-12 Sarl Ind Laser Partner Procede de realisation, sur un produit, d'un marquage invisible, pouvant etre revele
DE10004135B4 (de) 2000-01-31 2005-04-28 Ibidi Gmbh Kammer für Zellkulturen
NL1015686C2 (nl) * 2000-07-12 2002-01-15 Dsm Nv Werkwijze voor het met laserbestraling irreversibel aanbrengen van een voor het naakte oog onzichtbare markering op een polymeer vormdeel.
EP1343973B2 (fr) * 2000-11-16 2020-09-16 California Institute Of Technology Appareil et procedes pour effectuer des dosages et des criblages a haut rendement
DE10105711B4 (de) 2001-02-08 2005-03-10 Ibidi Gmbh Probenträger für chemische und biologische Proben
JP2002239484A (ja) * 2001-02-16 2002-08-27 Ushio Inc 誘電体バリア放電ランプを使った基板処理装置
JP2005504317A (ja) 2001-09-28 2005-02-10 イビディ ゲムベーハー フロー・チャンバー
US8206666B2 (en) * 2002-05-21 2012-06-26 Battelle Memorial Institute Reactors having varying cross-section, methods of making same, and methods of conducting reactions with varying local contact time
GB0212638D0 (en) 2002-05-31 2002-07-10 Ucb Sa Authentication means
GB0307615D0 (en) 2003-04-02 2003-05-07 Ucb Sa Authentication means
US7169615B2 (en) 2003-11-26 2007-01-30 General Electric Company Method of authenticating polymers, authenticatable polymers, methods of making authenticatable polymers and authenticatable articles, and articles made there from
JP4945893B2 (ja) * 2004-11-11 2012-06-06 大日本印刷株式会社 パターン形成用基板
ATE500895T1 (de) 2005-07-05 2011-03-15 Ibidi Gmbh Mikrofluid-vorrichtung und verfahren zur erzeugung diffusiv aufgebauter gradienten
DK1880764T3 (da) 2006-07-20 2012-12-17 Ibidi Gmbh Prøveholder til undersøgelse af cellevækst
FR2909922B1 (fr) * 2006-12-14 2010-08-13 Att Advanced Track & Trace Procede et dispositif de marquage d'objets et materiaux.
ATE523254T1 (de) * 2007-06-25 2011-09-15 Ibidi Gmbh Probenkammer
US20100186524A1 (en) * 2008-02-05 2010-07-29 Enertechnix, Inc Aerosol Collection and Microdroplet Delivery for Analysis

Also Published As

Publication number Publication date
US20110086420A1 (en) 2011-04-14
EP2292329B1 (fr) 2014-11-12
EP2292329A1 (fr) 2011-03-09
US9597688B2 (en) 2017-03-21

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