US6320196B1 — Multichannel high dynamic range scanner

Claim 1

Independent Claim 162 words

A multiple frequency laser induced fluorescence scanner for scanning a sample labeled with a plurality of luminescence dyes to cause fluorescence emission of light at different wavelengths, comprising: at least one laser for forming a first and second beam of light of at least first and second predetermined excitation wavelengths, each excitation wavelength preferentially exciting one of the plurality of luminescence dyes; an optical system for focusing the first and second beams of light on respective first and second spatially separated spots, and for collecting and transmitting the resulting emission through respective spatially separated apertures; and detector means coupled to the spatially separated apertures for measuring the emission signal level of the collected and transmitted emissions; wherein the optical system further comprises focusing lenses and a pair of optical fibers that transmit the spectral emission from the focusing lenses to the detector, wherein the pair of optical fibers act as spatially separated apertures for the respective emissions from the spatially separated spots.

Claim 2

Dependent Claim 162 words

A multiple frequency laser induced fluorescence scanner for scanning a sample labeled with a plurality of luminescence dyes to cause fluorescence emission of light at different wavelengths, comprising: at least one laser for forming a first and second beam of light of at least first and second predetermined excitation wavelengths, each excitation wavelength preferentially exciting one of the plurality of luminescence dyes; an optical system for focusing the first and second beams of light on respective first and second spatially separated spots, and for collecting and transmitting the resulting emission through respective spatially separated apertures; and detector means coupled to the spatially separated apertures for measuring the emission signal level of the collected and transmitted emissions; wherein the optical system further comprises focusing lenses and a pair of optical fibers that transmit the spectral emission from the focusing lenses to the detector, wherein the pair of optical fibers act as spatially separated apertures for the respective emissions from the spatially separated spots.

Claim 3

Dependent Claim 35 words

The multiple frequency laser induced fluorescence scanner as claimed in claim 1 , the detector means further comprising at least two photomultiplier tubes for separately processing the emission of a sample labeled with two dyes.

Claim 4

Dependent Claim 35 words

The multiple frequency laser induced fluorescence scanner as claimed in claim 1 , wherein the first and second beams of light are simultaneously focused on respective first and second separated spots by the same lens.

Claim 5

Dependent Claim 31 words

The multiple frequency laser induced fluorescence scanner as claimed in claim 1 , the detector means further comprising filters for reducing the overlap of the emission spectra detected to minimize crosstalk.

Claim 6

Dependent Claim 140 words

A multiple frequency laser induced fluorescence scanner for scanning a sample labeled with a plurality of luminescence dyes to cause fluorescence emission of light at different wavelengths, comprising: at least one laser for forming a first and second beam of light of at least first and second predetermined excitation wavelengths, each excitation wavelength preferentially exciting one of the plurality of luminescence dyes; an optical system for focusing the first and second beams of light on respective first and second spatially separated spots, and for collecting and transmitting the resulting emission through respective spatially separated apertures; and detector means coupled to the spatially separated apertures for measuring the emission signal level of the collected and transmitted emissions; wherein the first and second beams focused on the spots have orthogonal polarizations; the scanner additionally comprising polarizers each in a corresponding detection path.

Claim 7

Dependent Claim 140 words

A multiple frequency laser induced fluorescence scanner for scanning a sample labeled with a plurality of luminescence dyes to cause fluorescence emission of light at different wavelengths, comprising: at least one laser for forming a first and second beam of light of at least first and second predetermined excitation wavelengths, each excitation wavelength preferentially exciting one of the plurality of luminescence dyes; an optical system for focusing the first and second beams of light on respective first and second spatially separated spots, and for collecting and transmitting the resulting emission through respective spatially separated apertures; and detector means coupled to the spatially separated apertures for measuring the emission signal level of the collected and transmitted emissions; wherein the first and second beams focused on the spots have orthogonal polarizations; the scanner additionally comprising polarizers each in a corresponding detection path.

Claim 8

Independent Claim 174 words

A method for detecting fluorescence from a sample labeled with a plurality of fluorescent dyes, comprising: generating a plurality of laser beams of a first and second wavelength; directing the light of a first wavelength to a first spot and directing the light of a second wavelength to a second spot that is spatially separated from the first spot, to cause the sample to fluoresce and emit light at a plurality of different wavelengths; and scanning the spots across the sample in one direction and a slow scan direction; applying the resulting emission from the dye-labeled sample to a detector to generate an output signal representative of the fluorescence from the sample, wherein each detector only receives light from a spatially defined area illuminated by the laser that preferentially excites the dye to preferentially be detected by that detector; wherein a separation of the spots in the slow scan direction is provided to ensure that the excited dyes have fully recovered from any triplet saturation before they get scanned by the next laser beam.

Claim 9

Independent Claim 174 words

A method for detecting fluorescence from a sample labeled with a plurality of fluorescent dyes, comprising: generating a plurality of laser beams of a first and second wavelength; directing the light of a first wavelength to a first spot and directing the light of a second wavelength to a second spot that is spatially separated from the first spot, to cause the sample to fluoresce and emit light at a plurality of different wavelengths; and scanning the spots across the sample in one direction and a slow scan direction; applying the resulting emission from the dye-labeled sample to a detector to generate an output signal representative of the fluorescence from the sample, wherein each detector only receives light from a spatially defined area illuminated by the laser that preferentially excites the dye to preferentially be detected by that detector; wherein a separation of the spots in the slow scan direction is provided to ensure that the excited dyes have fully recovered from any triplet saturation before they get scanned by the next laser beam.

Claim 10

Dependent Claim 45 words

The method for detecting fluorescence from a sample labeled with a plurality of fluorescent dyes as claimed in claim 6 , the step of scanning more than one spot further comprising running the different laser beams through the same lens at a slightly different angle.

Claim 11

Independent Claim 162 words

A method for detecting fluorescence from a sample labeled with a plurality of fluorescent dyes, comprising: generating a plurality of laser light beams of a first and second wavelength; directing light of the first wavelength to a first spot and directing light of a second wavelength to a second spot that is spatially separated from the first spot, to cause the sample to fluoresce and emit light at a plurality of different wavelengths; scanning the spots across the sample; applying the resulting emission from the dye-labeled sample to a detector to generate an output signal representative of the fluorescence from the sample, wherein crosstalk is minimized as the emission spectra of the dyes detected overlap as little as possible and each detector only receives light from a spatially defined area illuminated by the laser that preferentially excites the dye to preferentially be detected by that detector; wherein the longest wavelength of the first and second wavelengths is scanned across the sample first.

Claim 12

Independent Claim 162 words

A method for detecting fluorescence from a sample labeled with a plurality of fluorescent dyes, comprising: generating a plurality of laser light beams of a first and second wavelength; directing light of the first wavelength to a first spot and directing light of a second wavelength to a second spot that is spatially separated from the first spot, to cause the sample to fluoresce and emit light at a plurality of different wavelengths; scanning the spots across the sample; applying the resulting emission from the dye-labeled sample to a detector to generate an output signal representative of the fluorescence from the sample, wherein crosstalk is minimized as the emission spectra of the dyes detected overlap as little as possible and each detector only receives light from a spatially defined area illuminated by the laser that preferentially excites the dye to preferentially be detected by that detector; wherein the longest wavelength of the first and second wavelengths is scanned across the sample first.

Claim 13

Dependent Claim 13 words

The method according to claim 8 wherein the dyes comprise R6G and Oxazine-1.

Claim 14

Dependent Claim 13 words

The method according to claim 8 wherein the dyes comprise Cy3 and Cy5.

Claim 15

Independent Claim 179 words

A method for detecting fluorescence from a sample labeled with a plurality of fluorescent dyes, comprising: generating a plurality of laser beams of a first and second wavelength; directing the light of a first wavelength to a first spot and directing the light of a second wavelength to a second spot that is spatially separated from the first spot, to cause the sample to fluoresce and emit light at a plurality of different wavelengths; and applying the resulting emission from the dye-labeled sample to a detector to generate an output signal representative of the fluorescence from the sample, wherein crosstalk is minimized as the emission spectra of the dyes detected overlap as little as possible and each detector only receives light from a spatially defined area illuminated by the laser that preferentially excites the dye to preferentially be detected by that detector; wherein up to two dyes are excited with orthogonal polarizations of light, such that fluorescence that is polarized parallel to the excitation light which preferentially excites a given dye is detected in the absence of depolarization effects.

Claim 16

Independent Claim 179 words

A method for detecting fluorescence from a sample labeled with a plurality of fluorescent dyes, comprising: generating a plurality of laser beams of a first and second wavelength; directing the light of a first wavelength to a first spot and directing the light of a second wavelength to a second spot that is spatially separated from the first spot, to cause the sample to fluoresce and emit light at a plurality of different wavelengths; and applying the resulting emission from the dye-labeled sample to a detector to generate an output signal representative of the fluorescence from the sample, wherein crosstalk is minimized as the emission spectra of the dyes detected overlap as little as possible and each detector only receives light from a spatially defined area illuminated by the laser that preferentially excites the dye to preferentially be detected by that detector; wherein up to two dyes are excited with orthogonal polarizations of light, such that fluorescence that is polarized parallel to the excitation light which preferentially excites a given dye is detected in the absence of depolarization effects.

Claim 17

Dependent Claim 24 words

The method of claim 11 , wherein the step of directing the light of a first wavelength and of a second wavelength, occurs simultaneously.

Claim 18

Dependent Claim 24 words

The method of claim 11 wherein the sample is a chip having different molecular counterparts in different areas of the chip which have hybridized.

Annuity Stage	Window Opens	Payment Deadline	Surcharge Period Until	Estimated Fee	Status
3.5 Years (Year 4)	2004-11-19	2005-05-21	2005-11-20	$2000.00	Window Closed / Lapsed
7.5 Years (Year 8)	2008-11-19	2009-05-21	2009-11-20	$3760.00	Window Closed / Lapsed
11.5 Years (Year 12)	2012-11-19	2013-05-21	2013-11-20	$7700.00	Window Closed / Lapsed

Multichannel high dynamic range scanner

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