Device for referencing fluorescence signals
First Claim
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1. A device for referencing fluorescence signals, comprising:
- an essentially non-fluorescent carrier;
a plurality of polymer layers applied to the carrier, at least one polymer layer applied to defined regions of the carrier; and
a plurality of biological probe molecules immobilized with respect to the carrier and configured to interact with a biological target molecule;
wherein;
the at least one polymer layer fluoresces in response to corresponding irradiation, and wherein at least two of the polymer layers differ with regard to their thickness and/or composition.
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Abstract
The invention relates to a device for referencing fluorescence signals and/or calibration of fluorescence detection systems, whereby the device comprises an essentially non-fluorescing support, on which are applied polymer layers in several defined regions and with partly varying thicknesses and/or compositions. Said polymer layers are applied to the support such as to fluoresce after corresponding irradiation and the device may thus be used as a fluorescence standard. The invention further relates to a method for the production of said fluorescence standards.
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Citations
53 Claims
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1. A device for referencing fluorescence signals, comprising:
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an essentially non-fluorescent carrier; a plurality of polymer layers applied to the carrier, at least one polymer layer applied to defined regions of the carrier; and a plurality of biological probe molecules immobilized with respect to the carrier and configured to interact with a biological target molecule; wherein; the at least one polymer layer fluoresces in response to corresponding irradiation, and wherein at least two of the polymer layers differ with regard to their thickness and/or composition. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 42, 43, 44, 45, 46)
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27. A method of manufacturing a device for referencing fluorescence signals, comprising:
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applying at least one polymer layer to at least one defined region of an essentially non-fluorescent carrier using a microtechnical process, wherein the at least one polymer layer fluoresces in response to corresponding irradiation; and immobilizing a plurality of biological probe molecules with respect to the carrier. - View Dependent Claims (28, 29, 30, 31, 32, 33)
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34. A method of online calibration of fluorescence detection systems with regard to at least one of their spatial or temporal resolution, their geometric and/or dynamic characteristics, and their characteristics caused by aging, comprising:
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irradiating a device including; an essentially non-fluorescent carrier; a plurality of polymer layers applied to the carrier, at least one polymer layer applied to defined regions of the carrier; and a plurality of biological probe molecules immobilized with respect to the carrier and configured to interact with a biological target molecule; wherein the at least one polymer layer fluoresces in response to corresponding irradiation, and wherein at least two of the polymer layers differ with regard to their thickness and/or composition; and measuring fluorescence from the at least one polymer layer. - View Dependent Claims (35, 36, 37, 38, 39)
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40. A method of cross-system and cross-device and/or cross-test comparison of cellular localization experiments and tissue slices which are evaluable by fluorescence microscopy, comprising:
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irradiating a device including; an essentially non-fluorescent carrier; a plurality of polymer layers applied to the carrier, at least one polymer layer applied to defined regions of the carrier, and a biological sample including a cell or a tissue slice immobilized with respect to the carrier; wherein the at least one polymer layer fluoresces in response to corresponding irradiation, and wherein at least two of the polymer layers differ with regard to their thickness and/or composition; and measuring fluorescence from the at least one polymer layer.
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41. A fluorescence calibration device, comprising:
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an essentially non-fluorescent carrier having one or more defined regions; at least one polymer layer applied to the one or more defined regions; and a plurality of biological probe molecules immobilized with respect to the carrier and configured to interact with a biological target molecule; wherein the polymer layer is uniform with regard to layer thickness and composition, such that the one or more defined regions fluoresce after corresponding irradiation.
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47. A device, comprising:
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a support; a plurality of spaced-apart polymer masses each being immobilized with respect to the support; and a plurality of biological probe molecules immobilized with respect to the support; wherein each of the polymer masses is configured to emit, at a common predetermined wavelength, a respective, different fluorescence intensity when irradiated with light; and
the biological probe molecules are configured to interact with a biological target molecule. - View Dependent Claims (48, 49, 50, 51)
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52. A device for referencing fluorescence signals, comprising:
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an essentially non-fluorescent carrier; and a plurality of polymer layers applied to the carrier, at least one polymer layer applied to defined regions of the carrier; and wherein; the at least one polymer layer fluoresces in response to corresponding irradiation, and wherein at least two of the polymer layers differ with regard to their thickness and/or composition, wherein the intensity of the fluorescence is adjustable by selecting one or more of the layer thickness and the composition of the polymer layers.
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53. A method of manufacturing a device for referencing fluorescence signals, comprising:
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applying at least one polymer layer to at least one defined region of an essentially non-fluorescent carrier using a microtechnical process, wherein the at least one polymer layer fluoresces in response to corresponding irradiation, wherein the at least one polymer layer is applied to the carrier by positive or negative photolithographic methods.
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Specification