Localization Microscopy Parallel Molecule Positioning
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Solution Overview
Problem
Current localization microscopy techniques, such as MINFLUX microscopy, are inefficient in determining the positions of multiple fluorophores in a sample, requiring a substantial amount of time.
Innovation Solution
A procedure using a localization microscope with a light modulator to create multiple light distributions with local intensity minima and adjacent intensity increase areas, allowing for the parallelized determination of molecule positions by independently positioning these light distributions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional localization microscopy techniques are used to determine positions of multiple fluorophores, then measurement precision is achieved, but productivity deteriorates due to substantial time requirements
Solution Approach 1:
The patent divides the field of view into multiple regions of interest, each assigned to a separate light distribution. This segmentation allows parallel processing of multiple fluorophores simultaneously, transforming a sequential measurement process into a parallel one, thereby improving productivity while maintaining measurement precision through the use of intensity minima for accurate localization
Solution Approach 2:
The patent introduces a spatial dimension by creating multiple light distributions at different positions simultaneously rather than scanning sequentially. By positioning multiple intensity minima across different spatial locations at the same time, the system can determine positions of multiple fluorophores in parallel, effectively adding a spatial parallelism dimension to the measurement process
2Productivity
If multiple light distributions are generated for parallel position determination, then productivity is improved, but device complexity increases
Solution Approach 1:
The patent employs a single light modulator that can generate multiple light distributions simultaneously, making this one device perform the function of what would traditionally require multiple separate scanning systems. The light modulator is controlled to create multiple intensity minima at different positions, enabling parallel measurement without proportionally increasing hardware complexity
Solution Approach 2:
The patent combines multiple light distributions into a single illumination system that operates simultaneously. By merging the functionality of multiple sequential light sources into one multi-functional illumination system, the patent achieves parallel processing capability while avoiding the complexity of multiple independent scanning systems
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 significantly reduces the time required to determine the positions of multiple fluorophores, enabling faster sample processing and improved efficiency in super-resolution microscopy.
Implementation Method 1
light distributions resulting from interference of coherent light are used to determine the positions of the molecules
Implementation Method 2
light distributions resulting from interference of coherent light
Implementation Method 3
The individual molecules are in a fluorescent state and are excited with excitation light to emit fluorescent light
Data Source
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AI summary
The invention relates to a method, to a computer program with instructions, and to a device for determining positions of two or more mutually spaced molecules in one or more spatial directions in a sample using a localization microscope. The invention additionally relates to a localization microscope which is used by a device according to the invention. In order to determine the positions of the molecules, light distributions are used which are produced on the basis of the interference of coherent light. In the method, a plurality of light distributions are generated (S1) using a first light modulator with a plurality of switchable pixels. The first light modulator is arranged on an image plane of the localization microscope. Each light distribution has a local intensity minimum and adjacent regions of increasing intensity. Each of the two or more molecules is illuminated (S2) with a respective light distribution. For each of the light distributions, photons emitted by the molecules are detected (S4) for different positions of the light distribution. In the process, the light distributions are positioned (S3) independently of one another, and the positions of the molecules are finally derived (S5) from the photons detected for the different positions of the light distributions.