Apparatus for quantitative imaging of multiple fluorophores
OwnerNils R. D. Aslund, Kjell S. Carlssoninventors
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Abstract
A quantitative fluorometer for multiple fluorophores having a separate time-modulated beam of excitation light for each fluorescent target. Each beam is synchronized with a separate lock-in amplifier. The fluorophores are simultaneously excited and the combined fluorescent emission is resolved into components corresponding to each fluorophore. Confocal scanning means are used to excite and detect fluorescent emission from locations throughout a volume. The location specific output of each amplifier is stored in a computer which resolves the emission into the components corresponding to each fluorophore. The location specific data may be further processed or visually displayed.
Claims23
A device for quantifying one or more fluorescent targets by excitation with electromagnetic radiation of multiple discrete wavelengths comprising:
- means for directing dual beams of continuous electromagnetic radiation, each beam having a discrete wavelength, said beams impinging on a single location where fluorescent targets are present causing excitation thereof;
- means for modulating the intensity of each of said beams at separate modulation frequencies to give each of said beams a specific time modulated waveform;
- means for detecting the combined fluorescent emission from said targets in response to said excitation;
- and
- demodulation means for extracting the contributions corresponding to each of said targets from said detected combined fluorescent emission, thereby quantifying said targets.
A system for performing quantitative microfluorometry comprising:
- confocal means for directing dual beams of light, each of said beams having a discrete wavelength, said beams impinging on a location where fluorescent targets are present causing excitation thereof;
- means for modulating each of said beams with a separate modulation frequency;
- scanning means having a sampling rate for detecting the combined fluorescent emission from said targets at locations in a scanning plane or planes in response to said excitation;
- phase and frequency discriminating means for extracting the individual emissions corresponding to each of said targets from said detected combined fluorescent emission, thereby quantifying said targets;
- means for converting the discriminated emissions from said targets to location specific signals;
- and
- means for storing said location specific signals in a memory.
Dependencies are inferred from the claim text.