3D Localization Microscopy Phase Modulation Z-Axis Determination
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Solution Overview
Problem
Current 3D localization microscopy techniques are limited by the optical diffraction limit and struggle to accurately determine the z-axis position of emitters without moving the sample or objective, and they do not effectively incorporate additional dimensions or properties of light emitters.
Innovation Solution
The method involves creating a phase difference between light passing through different parts of the objective to produce a point emitter image with two lobes, where the separation of these lobes is related to the emitter's position and potentially other properties like spectral content, lifetime, or polarization, allowing for improved z-axis determination and additional dimension encoding in 4D localization microscopy.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional 3D localization microscopy methods are used, then x and y axis location information can be determined beyond the diffraction limit, but z axis position determination requires moving the sample or objective which reduces productivity and increases complexity
Solution Approach 1:
The patent introduces a fourth dimension by encoding an additional property of the light emitter (such as spectral content, lifetime, or polarization) alongside the spatial position. This is achieved by causing a phase difference between light passing to or from one part of the objective relative to light passing to or from another part of the objective, producing a point emitter image with two lobes where the separation encodes both z-position and another emitter property, eliminating the need for mechanical scanning
Solution Approach 2:
The patent changes the optical parameters by engineering the point spread function (PSF) to produce two separated image spots whose separation and orientation depend on the emitter's z-position and another property. By calibrating the system to recognize the rotational behavior and separation characteristics of these spots, the method estimates both z-position and the additional property from a single 2D image without mechanical movement
2Productivity
If the point spread function is engineered to encode 3D position information in the lateral PSF profile shape, then z position can be determined from a single image, but the method does not encode additional emitter properties
Solution Approach 1:
The patent extends conventional 3D localization by adding a fourth dimension that encodes an additional property of the light emitter. The engineered PSF produces two lobes where the separation between lobes encodes the z-position, while another property of the image (such as relative brightness, spectral content, lifetime, or polarization) encodes the additional emitter property, allowing simultaneous determination of both from a single 2D image
Solution Approach 2:
The patent creates a multi-functional PSF that simultaneously encodes multiple pieces of information: the spatial position (x, y, z) and an additional emitter property (such as spectral content, lifetime, or polarization). The two-lobe PSF structure serves multiple functions by encoding both positional and non-positional information in a single imaging measurement, making the system universally applicable to various emitter properties
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 enhances the resolution and accuracy of 3D imaging by enabling precise z-axis determination and encoding additional properties, surpassing the limitations of existing methods, particularly in achieving sub-diffraction limit resolution and maintaining robustness across various imaging conditions.
Implementation Method 1
causing a phase difference between light passing to or from one part of the objective relative to light passing to or from another part of the objective to produce a point emitter image which comprises two image lobes
Implementation Method 2
The microscope resolution is limited by the optical diffraction limit of the microscope. Diffraction causes a single point emitter of a size less than the diffraction limit of the microscope to be imaged as a larger spot
Data Source
AI summary
A 3D localization microscopy system, 4D localisation microscopy system, or an emitter tracking system arranged to cause a phase difference between light passing to or from one part of the objective relative to light passing to or from another part of the objective, to produce a point emitter image which comprises two lobes, a separation between which is related to the position of the emitter relative to the objective of the imaging system, and in the 4D system a further property of the image or of the light to or from the objective is related to another location independent property of the emitter.