Multiple-label fluorescence imaging using excitation-emission matrices
First Claim
1. An apparatus, comprising:
- a light source for producing spectral excitation light having wavelength variation along a first dimension across a spectral range;
a scanning device for scanning the spectral excitation light across a sample substantially parallel to the first dimension such that the wavelength variation across the spectral range of the spectral excitation light interacts with a spatial position along the sample to emit a secondary light; and
an imaging device for capturing the secondary light as a spectral emission light having wavelength variation along a second dimension.
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Abstract
Methods and devices are disclosed which apply an excitation-emission matrix (EEM) to a heterogeneous, two-dimensional sample, allowing a considerably larger number of emitting, e.g. fluorescent, labels to be used simultaneously. This may be accomplished by employing a spectroscopic method of excitation-emission matrices which allows discrimination of species with similar emission spectra, and also allows positive identification of energy transfer between emitting species. The methods and devices may employ a novel excitation-light scanning technique which allows imaging of the emission from the heterogeneous sample both in two spatial dimensions (length and width) and in two spectral dimensions (excitation and emission wavelength). This light scanning technique maximizes the throughput of excitation light, increasing the sensitivity and hence the reading speed of the instrument.
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Citations
23 Claims
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1. An apparatus, comprising:
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a light source for producing spectral excitation light having wavelength variation along a first dimension across a spectral range; a scanning device for scanning the spectral excitation light across a sample substantially parallel to the first dimension such that the wavelength variation across the spectral range of the spectral excitation light interacts with a spatial position along the sample to emit a secondary light; and an imaging device for capturing the secondary light as a spectral emission light having wavelength variation along a second dimension. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11)
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12. A method, comprising:
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producing spectral excitation light having wavelength variation along a first dimension across a spectral range; scanning the spectral excitation light across a sample substantially parallel to the first dimension such that the wavelength variation across the spectral range of the spectral excitation light interacts with a spatial position along the sample to emit a secondary light; and capturing the secondary light as a spectral emission light having wavelength variation along a second dimension. - View Dependent Claims (13, 14, 15, 16, 17, 18, 19, 20, 21, 22)
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23. An apparatus, comprising:
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means for producing spectral excitation light having wavelength variation along a first dimension across a spectral range; means for scanning the spectral excitation light across a sample substantially parallel to the first dimension such that the wavelength variation across the spectral range of the spectral excitation light interacts with a spatial position along the sample to emit a secondary light; and means for capturing the secondary light as a spectral emission light having wavelength variation along a second dimension.
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Specification