Dual-Emitter Tracking Using MINFLUX Intensity Profile Localization
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Solution Overview
Problem
Existing microscopy techniques struggle to simultaneously track two optically distinguishable light-emitting particles or units that are closely adjacent, with high spatial and temporal resolution, due to limitations in spatial-temporal resolution and interference from marker size and crosstalk.
Innovation Solution
A method using MINFLUX or STED-MINFLUX techniques with overlapping excitation and emission suppression light distributions, combined with iterative localization and pulsed excitation, allows simultaneous tracking of closely adjacent emitters by generating multiple intensity profiles and assigning emissions to these profiles during data acquisition.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Device complexity
If a single receiver is used to detect electromagnetic signals from multiple emitters, then the device complexity is reduced, but the ability to simultaneously track multiple emitters deteriorates due to signal mixing and inability to distinguish individual emitter characteristics
Solution Approach 1:
The patent divides the detection task by assigning different frequency bands to different receivers. Each receiver is tuned to a specific frequency range, allowing simultaneous detection of multiple emitters without signal mixing. This segmentation of the frequency spectrum enables each receiver to independently track emitters within its designated band, resolving the contradiction between simple device structure and precise multi-emitter tracking.
Solution Approach 2:
The patent introduces frequency as an intermediary parameter that mediates between multiple emitters and the receivers. By assigning unique frequency signatures to different emitters and matching receivers to specific frequency ranges, the system enables simultaneous tracking without direct signal interference. This intermediary frequency domain approach allows the simple single-receiver architecture to effectively handle multiple emitters.
2Measurement precision
If multiple receivers are used to track multiple emitters simultaneously, then the emitter tracking capability is improved, but the device complexity and cost increase due to additional hardware components
Solution Approach 1:
The patent makes each receiver universal within its frequency band by designing it to detect signals from any emitter operating in that band. Rather than having dedicated receivers for each emitter, the system uses multiple receivers that can each independently track multiple emitters within their respective frequency ranges. This multi-functionality reduces the total number of receivers needed while maintaining precise tracking capability.
Solution Approach 2:
The patent transitions from spatial separation of receivers to frequency domain separation. Instead of positioning multiple receivers in different physical locations to track different emitters, the system assigns emitters to different frequency dimensions. This dimensional change in the signal space allows a simpler receiver architecture to achieve multi-emitter tracking by operating in the frequency domain rather than relying on complex spatial arrangements.
3Adaptability or versatility
If the receiver operates across a broad frequency range to detect all emitters, then the adaptability is improved, but the signal detection precision deteriorates due to frequency mixing and interference
Solution Approach 1:
The patent segments the broad frequency range into multiple non-overlapping sub-bands, with each receiver dedicated to a specific sub-band. This segmentation prevents frequency mixing and interference that would occur if a single receiver tried to process the entire broad spectrum simultaneously. Each receiver achieves high detection precision within its narrow band while the collective system maintains broad frequency coverage through the segmented approach.
Solution Approach 2:
The patent applies local quality by optimizing each receiver for its specific frequency sub-band rather than designing a single receiver to handle all frequencies equally. Each receiver is tuned and configured with characteristics optimized for its local frequency region, enabling high detection accuracy in that specific band. This localized optimization resolves the contradiction between broad adaptability and precise detection by making each component specialized for its local operating conditions.
Data Source
AI summary
The object of the invention is to provide solutions that allow two optically distinguishable light-emitting particles or light-emitting units which are closely adjacent to be simultaneously tracked with high spatial and temporal resolution. The object is achieved by means of MINFLUX methods or STED-MINFLUX methods or a combination of both method types. Either a location of the first emitter or locations of the first and second emitters or a common mean location of both emitters is determined on the basis of various profiles of intensity increase regions of an excitation light or of an emission prevention light and associated measurement values of the emission. This is repeated in temporal sequence, in each case with adaptations of the profiles of intensity increase regions to the current positions of the emitters.