Method and device for conducting the spectral differentiating, imaging measurement of fluorescent light
First Claim
1. A method for an image producing measurement of fluorescent light, comprising:
- irradiating a sample, containing fluorophores of at least one species, with exciting light of at least one excitation channel defined by its spectral properties, andreceiving the fluorescent light emitted by the sample by at least one detection channel defined by its spectral detection characteristic and converting it into a digital signal,storing the digital signal in a memory unit of a digital data processing unit, andproducing an image or image file of the sample based on the digital signal,wherein, prior to carrying out a measurement, properties of the at least one excitation channel and the at least one detection channel are predetermined according to a result of a mathematical optimization process carried out by a computing unit of the digital data processing unit, which takes into consideration fluorescence characteristics of at least one of the fluorophores presumed by a user to be in the sample,whereinthe optimization process includesa) setting up a linear system of equations,coefficients of which represent spectral characteristics of measurement channels, which are respectively defined as a combination of one excitation channel and one detection channel, andwhich describes a relationship between a user-presumed chemical composition of the sample and a signal calculated based on the measurement channel characteristics to be optimized,b) varying the coefficients, until a unique solution to the equation system becomes possible,c) the coefficients calculated as a result of the optimization process forming a basis of the predetermination of the properties of the excitation and detection channels.
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Abstract
A method and a device for image producing measurement of fluorescent light, according to which a sample containing fluorophores of different species is irradiated with excitation light of at least one excitation channel defined by its spectral properties. The fluorescent light emitted by the sample is received by at least one detection channel defined by its spectral detection characteristic, and is converted into a digital signal, which is stored for further processing The properties of a number of measuring channels, respectively defined as specific combinations consisting of an excitation channel and a detection channel, are automatically set before conducting the measurement according to the result of a mathematical optimization process, which takes into account the fluorescence characteristics of at least some of the fluorophores presumed by the user to be in the sampler.
14 Citations
19 Claims
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1. A method for an image producing measurement of fluorescent light, comprising:
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irradiating a sample, containing fluorophores of at least one species, with exciting light of at least one excitation channel defined by its spectral properties, and receiving the fluorescent light emitted by the sample by at least one detection channel defined by its spectral detection characteristic and converting it into a digital signal, storing the digital signal in a memory unit of a digital data processing unit, and producing an image or image file of the sample based on the digital signal, wherein, prior to carrying out a measurement, properties of the at least one excitation channel and the at least one detection channel are predetermined according to a result of a mathematical optimization process carried out by a computing unit of the digital data processing unit, which takes into consideration fluorescence characteristics of at least one of the fluorophores presumed by a user to be in the sample, wherein the optimization process includes a) setting up a linear system of equations, coefficients of which represent spectral characteristics of measurement channels, which are respectively defined as a combination of one excitation channel and one detection channel, and which describes a relationship between a user-presumed chemical composition of the sample and a signal calculated based on the measurement channel characteristics to be optimized, b) varying the coefficients, until a unique solution to the equation system becomes possible, c) the coefficients calculated as a result of the optimization process forming a basis of the predetermination of the properties of the excitation and detection channels. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19)
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Specification