Multiple inclined beam line-scanning imaging apparatus, methods, and applications
AssigneeUniversity of Central Florida Research Foundation, Inc.
InventorsKyu Young Han, Jialei Tang
Life of this patent
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Abstract
A dual inclined beam line-scanning confocal microscope apparatus, associated method, and applications thereof. An embodied dual-inclined beam line-scanning confocal microscope (2iLS) utilizes dual, parallel excitation beams each having a focused line shape that are scanned over a fluorescent sample. The emitted fluorescence from the sample is spatially filtered and detected by an array detector. 2iLS microscopy provides high resolution, ultrasensitivity, and deep optical sectioning capability. A reduced excitation intensity lowers photobleaching and photodamage.
Claims12
A dual-inclined beam line-scanning (2iLS) microscope, comprising:
- an optical train having a Cartesian z-direction optical axis, said optical train including a first optical component/assembly disposable to receive at least one collimated sample object fluorescence excitation wavelength input light beam, wherein said first optical assembly has optical characteristics suitable to transform the collimated input light beam into at least two parallel collimated output line beams oriented in an x-y plane that is orthogonal to the optical z-axis and characterized by a length, Ly, a width, wx, and a thickness, dz;
- a second optical component/assembly disposable to receive the at least two parallel collimated output line beams and having optical characteristics suitable to orient the at least two parallel collimated output line beams in the x-y plane at an angle, θ, with respect to the optical z-axis;
- a third optical component/assembly having optical characteristics suitable to focus the at least two parallel inclined line beams at an image plane;
- a fourth optical component/assembly disposed in a conjugate back focal plane of the image plane having optical characteristics suitable to sweep the at least two parallel inclined line beams along a horizontal x-axis in the image plane;
- and
- a detector/camera assembly disposable in a conjugate image plane including either a confocal slit or a functionally equivalent electrooptical confocal slit to receive a fluorescence emission from the sample object,
- wherein said 2iLS microscope is to be configured as a confocal, fluorescence emission microscope.
A method for imaging a sample object, comprising:
- generating at least two parallel line beams having a wavelength suitable for generating a fluorescence emission from a region of the sample object, wherein the at least two parallel line beams have a length, Ly, thickness, dz, and width, wx, wherein Ly and wx are measured in an x-y Cartesian plane and dz is measured in an orthogonal z-axis beam propagation direction;
- disposing the at least two parallel line beams at an angle, θ, in the x-y Cartesian plane with respect to the z-axis beam propagation direction;
- sweeping the inclined at least two parallel line beams across a region of the sample object and generating a fluorescence emission from the sample object;
- propagating the fluorescence emission from the sample object to a detector comprising a functional confocal slit, wherein the at least two parallel line beams are synchronously swept with an output of the detector.
Dependencies are inferred from the claim text.