Spectrally-resolving 3D Localization Microscopy via Anisotropic Distortion
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Solution Overview
Problem
Current localization microscopy techniques face challenges in achieving both spectral and depth resolution without photon suppression, particularly in three-dimensional imaging, where existing methods often result in unequal intensity between image lobes for different wavelengths, leading to reduced resolution and increased measurement time.
Innovation Solution
A localization microscope with an optical manipulation device that uses two anisotropy elements with non-parallel anisotropy axes, each neutral for a specific wavelength, to create rotationally asymmetric image lobes whose relative locations depend on the emitter's depth location and wavelength, allowing for simultaneous depth and spectral coding without photon suppression.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional localization microscopy is used for three-dimensional imaging, then depth resolution can be achieved, but spectral resolution is lost due to photon suppression
Solution Approach 1:
The detection space is segmented into multiple detection spaces corresponding to different wavelengths. The optical manipulation device separates sample light of different wavelengths into different detection spaces, allowing simultaneous detection of spectral and depth information without photon suppression.
Solution Approach 2:
The patent adds spectral dimension to the existing depth encoding. By creating wavelength-dependent anisotropic distortions, the system encodes both depth position and wavelength information in the spatial distribution of image lobes, transforming a 1D depth problem into a 2D spatial-spectral problem.
2Measurement precision
If wavelength-dependent anisotropic distortions are applied to achieve spectral resolution, then spectral information can be detected, but unequal intensity between image lobes occurs for different wavelengths
Solution Approach 1:
Each detection space is optimized with specific optical parameters (magnification, numerical aperture) tailored to its wavelength range. This local optimization compensates for wavelength-dependent intensity variations, ensuring uniform detection sensitivity across all spectral channels.
Solution Approach 2:
The patent adjusts optical parameters such as magnification and numerical aperture for different detection spaces to compensate for intensity variations. By changing these parameters wavelength-dependently, the system maintains equal intensity between image lobes across different spectral channels.
3Measurement precision
If multiple detection spaces with different optical parameters are used for spectral detection, then spectral resolution is achieved, but device complexity increases
Solution Approach 1:
A single optical manipulation device performs multiple functions: it simultaneously encodes depth position, separates wavelengths, and directs them to appropriate detection spaces. This multi-functionality reduces the need for separate spectral separation components, simplifying the overall system architecture.
Solution Approach 2:
The optical manipulation device automatically routes different wavelengths to their corresponding detection spaces based on their spectral properties. This self-organizing behavior eliminates the need for complex external control systems or additional spectral separation components.
4Measurement precision
If sequential imaging in different color channels is used, then spectral resolution can be achieved, but measurement time increases significantly
Solution Approach 1:
The patent merges multiple spectral channels into a single simultaneous detection process. By creating multiple detection spaces that operate in parallel within the same image plane, the system captures all spectral information in one exposure, eliminating sequential imaging delays.
Solution Approach 2:
All spectral channels are detected simultaneously and continuously in a single measurement process. The optical manipulation device maintains continuous detection across all wavelengths without interruption or sequential switching, maximizing measurement efficiency.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This approach enables the achievement of both spectral and depth resolution without photon suppression, allowing for accurate decoding of wavelength and depth information from the anisotropically distorted point emitter images, thereby enhancing the resolution and efficiency of three-dimensional localization microscopy.
Implementation Method 1
a first and a second anisotropy element are used in the optical manipulation device that anisotropically influence in each case the point spread function of the imaging to produce the image lobes of the point emitter image
Data Source
AI summary
A localization microscope including an imaging device emitting sample light from a focal plane into an image plane, including an optical-manipulation device for depth-dependent influencing of a point-spread function of the imaging and influencing the point-spread function of the imaging such that a point emitter is imaged in the image plane into an image that is rotationally asymmetrically distorted. A form of the distortion depends on the location of the point emitter with respect to the focal plane and a wavelength of the sample light. The optical manipulation device includes first and second anisotropy elements that anisotropically influence the point spread function to produce rotational asymmetry of the point emitter image. The elements are arranged one behind the other in the imaging direction, with anisotropy axes at an angle to one another. Both elements have differing neutral wavelength at which they do not anisotropically influence the point spread.