High-speed multiple wavelength illumination source, apparatus containing the same, and applications thereof to methods of irradiating luminescent samples and of quantitative luminescence ratio microscopy
First Claim
1. A vibration-free method of irradiating a sample, comprising the steps of:
- (A) irradiating said sample with light having a first wavelength, a first intensity or both a first wavelength and a first intensity, by passing an electric current having a first value through a first arc and passing an electric current having a second value through a second arc, said first value being greater than said second value; and
within a period of time of from 1 nanosecond (1 ns) to 1 second (1 s) of said irradiating step (A),(B) irradiating said sample with light having a second wavelength, a second intensity or both a second wavelength and a second intensity by reducing the value of said electric current passed through said first light source to a third value and increasing said electric current through said second light source to a fourth value, said fourth value being greater than said third value,wherein each of said arcs has a wattage of from 10 to 2000 W.
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Abstract
A high-speed, non-mechanical multiple wavelength illumination source is provided, which contains a first light source, a second light source providing a different wavelength of light, and a controlling means for controlling electric current to the first and second light sources. The illumination source is particularly useful in an improved apparatus for quantitative luminescence ratio photometry and/or luminescence ratio imaging. Methods of luminescence emission ratio photometry and luminescence emission ratio imaging using the illumination source and apparatus are also provided. The present invention offers particular advantages in photomicroscopy and in microscopic imaging.
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Citations
21 Claims
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1. A vibration-free method of irradiating a sample, comprising the steps of:
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(A) irradiating said sample with light having a first wavelength, a first intensity or both a first wavelength and a first intensity, by passing an electric current having a first value through a first arc and passing an electric current having a second value through a second arc, said first value being greater than said second value; and within a period of time of from 1 nanosecond (1 ns) to 1 second (1 s) of said irradiating step (A), (B) irradiating said sample with light having a second wavelength, a second intensity or both a second wavelength and a second intensity by reducing the value of said electric current passed through said first light source to a third value and increasing said electric current through said second light source to a fourth value, said fourth value being greater than said third value, wherein each of said arcs has a wattage of from 10 to 2000 W. - View Dependent Claims (2, 3, 4, 5)
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6. An apparatus for sequentially irradiating a sample with two or more wavelengths, intensities, or wavelengths and intensities of light, comprising:
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(A) a first arc which generates a first light beam having a first wavelength, a first intensity or both a first wavelength and a first intensity; (B) a second arc which generates a second light beam having a second wavelength, a second intensity or both a second wavelength and a second intensity, wherein said second wavelength differs from said first wavelength, said second intensity differs from said first intensity, or said second wavelength and said second intensity differ from said first wavelength and said first intensity; and (C) a controlling means for controlling electric current to each of said first and second arcs, said controlling means being electrically connected to each of said first and second arcs, wherein each of said first and second arcs has a wattage of from 10 to 2000 W. - View Dependent Claims (7, 8, 9, 10, 11, 12, 13, 14)
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15. An apparatus for luminescence emission photometry, comprising:
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(a) a first arc which generates a first light beam having sufficient energy to electronically excite at least one luminescent compound in a sample, (b) a second arc which generates a second light beam having sufficient energy to electronically excite at least one luminescent compound in said sample, (c) a controlling means for controlling electric current to each of said first and second arcs, said controlling means being electrically connected to each of said first and second arcs, (d) a first vibration-free means for selecting a wavelength of light, positioned in the path of said first light beam to provide said first light beam with a first wavelength, (e) a second vibration-free means for selecting a wavelength of light, positioned in the path of said second light beam to provide said second light beam with a second wavelength, said second wavelength differing from said first wavelength, (f) a mounting platform for said sample, positioned such that each of said first and second light beams strikes said sample so that said luminescent compound produces a luminescent emission, and (g) a detector positioned to receive said luminescent emission, wherein each of said first and second arcs has a wattage of from 10 to 2000 W. - View Dependent Claims (16, 17, 18, 19)
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20. A vibration-free method of determining the concentration of a luminescence-affecting chemical species in a sample capable of emitting luminescent radiation, comprising the steps of:
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(A) irradiating said sample with light having a first wavelength and having sufficient energy to cause said sample to emit a first luminescent emission by passing an electric current having a first value through a first arc and passing an electric current having a second value through a second arc, said first value being greater than said second value; then, within a period of time of from 1 ns to 1 s, (B) irradiating said sample with light having a second wavelength and having sufficient energy to cause said sample to emit a second luminescent emission by reducing the value of said electric current passed through said first light source to a third value and increasing said electric current through said second light source to a fourth value, said fourth value being greater than said third value, (C) measuring the intensity of each of said first and second luminescent emissions, (D) determining a ratio of one of said first and second luminescent emission intensities to the remaining one of said first and second luminescent emission intensities, and (E) correlating said ratio to said concentration of said luminescence-affecting chemical species in said sample, wherein each of said arcs has a wattage of from 10 to 2000 W. - View Dependent Claims (21)
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Specification