Fluorescence imaging system employing a macro scanning objective
First Claim
1. An optical scanning system for microscopy of a sample comprising,a photodetector,a beam source for emitting an incident beam,a telecentric objective disposed proximate to said sample, defining an optical axis, a high numerical aperture and a single pupil, with said single pupil located external to said objective in said optical axis, said objective being positioned to receive said incident beam and directing said incident beam onto said sample, producing an illuminated spot thereon with said objective collecting light emitted from said spot and forming a retro-beam, said objective being afocal at said external pupil,means, positioned at said pupil, for scanning said spot, on said sample surface, with substantially all light associated with said retro-beam impinging upon said scanning means, andmeans positioned in said optical axis between said beam source and said objective for separating said incident beam from said retro-beam, wherein said objective directs said retro-beam onto said separating means, with said separating means directing said retro-beam onto said photodetector.
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Accused Products
Abstract
A fluorescence imaging system that includes an objective that is both achromatic and has an external entrance pupil. The objective also serves as a condenser for the system which substantially reduces the system'"'"'s cost and footprint. With the objective positioned above a sample so that they are in close proximity to one another, a laser directs a collimated beam of light to a scan device located at the objective entrance pupil. The scan device reflects, refracts, or diffracts the light through the lens to illuminate a spot on the sample'"'"'s surface. The scan device illuminates a line or an area on the sample surface by varying the angle of laser light, in one or two dimensions, into the objective. The sample emits fluorescent light in response to the illumination. The fluorescence light is collected by the objective and passes through the system along the path of the illumination light. A wavelength-discriminating dichroic filter is placed along the optical axis between the laser and the objective to direct the fluorescent light onto a photo-detector to produce a signal representing the sample surface emitting the fluorescent light. A display device is provided that displays the digitized data in a raster format.
134 Citations
39 Claims
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1. An optical scanning system for microscopy of a sample comprising,
a photodetector, a beam source for emitting an incident beam, a telecentric objective disposed proximate to said sample, defining an optical axis, a high numerical aperture and a single pupil, with said single pupil located external to said objective in said optical axis, said objective being positioned to receive said incident beam and directing said incident beam onto said sample, producing an illuminated spot thereon with said objective collecting light emitted from said spot and forming a retro-beam, said objective being afocal at said external pupil, means, positioned at said pupil, for scanning said spot, on said sample surface, with substantially all light associated with said retro-beam impinging upon said scanning means, and means positioned in said optical axis between said beam source and said objective for separating said incident beam from said retro-beam, wherein said objective directs said retro-beam onto said separating means, with said separating means directing said retro-beam onto said photodetector.
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24. An optical scanning system for microscopy of a sample, said system comprising,
a source to produce an incident beam, a photodetector, a telecentric objective lens means for directing said incident beam onto said sample forming an illuminated spot thereon, and collecting light emitted from said spot, forming a retro-beam, said objective lens means defining a single pupil and a working distance, with said pupil being located external to said objective lens means, said objective lens means being afocal at said pupil, said working distance being measured between said sample and said objective lens means, with said objective lens means adapted to provide different numerical apertures, with said numerical apertures being inversely proportional to said working distance so as to maintain said objective lens means proximate to said sample, means, in said optical axis at said pupil, for scanning said spot across said sample, and means, positioned in said optical axis between said source and said objective lens means, for separating said incident beam from said retro-beam, wherein said objective lens means directs said retro-beam toward said photodetector.
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37. An optical scanning system for microscopy of a sample, said system comprising,
a beam source for emitting a collimated beam of coherent light, a photodetector, a telecentric objective defining an optical axis and a single pupil, with said pupil located external to said objective, said objective positioned to receive said collimated beam therethrough to illuminate a spot on said sample and collect light emitted from said spot, and forming a retro-beam, said objective being achromatic and afocal at said pupil, means, in said optical axis, for scanning said spot, on said sample, said scanning means including a reflective element positioned at said pupil of said objective, with said telecentric objective adapted to form said retro-beam with a cross-sectional area of sufficient size relative to said reflective element to ensure substantially all light associated with said retro-beam impinges upon said reflective element, and means positioned in said optical axis between said beam source and said objective for separating said incident beam from said retro-beam, wherein said objective directs said retro-beam toward said photo-detector, with said photo-detector producing signals representing said substantially all light emitted from said spot.
Specification