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US 9,791,371 B2Granted
Systems and methods for distinguishing stimulated emissions as a means of increasing the signal of fluorescence microscopy
AssigneeThe United States of America, as represented by the Secretary, Department of Health and Human Services
InventorsAndrew York, Sanjay Varma
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Abstract
Embodiments of a fluorescence microscopy system that employs a technique for distinguishing stimulated emission as a means for enhancing signal strength of fluorescent markers are disclosed.
Claims17
A fluorescence microscopy system comprising:
- an excitation source for generating an excitation light beam along a first axis;
- a stimulation source for generating a stimulation light beam along a second axis that is in perpendicular orientation relative to the first axis, wherein the excitation light beam and the stimulation light beam have different wavelengths;
- a sample having one or more fluorescent markers which are illuminated by the excitation light beam such that the one or more fluorescent markers assumes an excited state and then illuminated again by the stimulation light beam to generate a stimulated emission by the initially excited sample;
- an objective lens oriented along the second axis for capturing and focusing the stimulation light beam and stimulated emission emitted by the one or more fluorescent markers;
- a stimulation light beam block component positioned at a focal point of the objective lens such that the stimulation light beam is blocked while allowing the stimulated emission to pass;
- and
- a detector for detecting the stimulated emission.
4 more elements
A method for fluorescence microscopy comprising:
- illuminating one or more fluorescent markers in a sample with an excitation light beam from an excitation source oriented along a first axis such that the one or more fluorescent marker assume an excited state;
- illuminating the one or more fluorescent markers in the excited state with a stimulation light beam from a stimulation source oriented along a second axis that is in perpendicular relation to the first axis, wherein a stimulated emission is generated by the one or more fluorescent markers when illuminated by the stimulation light beam, wherein the stimulated light beam and the excitation light beam have different wavelengths;
- focusing the stimulation light beam and the stimulated emission through an objective lens;
- positioning a stimulated light beam block component along the second axis such that the stimulated light beam is focused onto and blocked by the stimulated light beam block component, and wherein the stimulated emission is not blocked by the positioning of the stimulated light beam block;
- and
- detecting the stimulated emission by a detector.
3 more elements
A fluorescence microscopy system comprising:
- an excitation source for generating an excitation light beam along a first axis;
- a stimulation source for generating a stimulation light beam along a second axis that is in perpendicular relation relative to the first axis, wherein the excitation light beam and the stimulation light beam have different wavelengths;
- a combined excitation/stimulation light beam that is generated by the intersection of the excitation light beam with the stimulation light beam;
- an excitation objective lens oriented along the first axis for capturing the combined excitation/stimulation light beam;
- a sample having one or more fluorescent markers in a focal plane of the excitation objective lens, wherein the one or more fluorescent markers are illuminated by the combined excitation/stimulation light beam for producing an stimulated emission;
- a detection objective lens oriented in perpendicular orientation relative to the excitation objective for capturing and focusing the stimulated emission emitted by the one or more fluorescent markers of the sample;
- and
- a detector for detecting the stimulated emission from the detection objective lens.
5 more elements
Dependencies are inferred from the claim text.