Non-congruent Detector Elements for MINFLUX Background Correction
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Solution Overview
Problem
Current MINFLUX methods face challenges in accurately correcting for background emissions, especially when they are inhomogeneously distributed, leading to measurement errors and inefficiencies in determining the position of emitters in a sample.
Innovation Solution
The method employs a plurality of detector elements with non-congruent active areas to detect light emissions and background separately, allowing for location-dependent background correction and improved position estimation by determining specific background values for each illumination step, thereby enhancing the separation of signal from background light.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If a single detector element is used to detect light emissions in MINFLUX methods, then the device complexity is reduced, but the measurement precision of emitter position deteriorates due to inability to correct inhomogeneous background emissions
Solution Approach 1:
The detector is divided into multiple detector elements (e.g., pixels in a camera or elements in a SPAD array), each capable of independently detecting light emissions. This segmentation allows the system to capture spatially resolved information about both emitter signals and background emissions, enabling background subtraction and improved position accuracy without requiring complex mechanical scanning systems.
Solution Approach 2:
The invention transitions from point-by-point temporal scanning to simultaneous spatial detection across multiple detector elements. By capturing light emissions across a two-dimensional detector array, the system obtains spatial information about background distributions that can be used to correct position measurements, effectively adding a spatial dimension to the measurement process.
2Loss of information
If background correction is performed using conventional methods with congruent detector element projections, then the device complexity is minimized, but the loss of information increases due to inability to separate signal from inhomogeneous background
Solution Approach 1:
Each detector element is assigned a unique projection geometry into the sample plane, creating local quality differences across the detector array. This allows each element to sample different portions of the light field, including both emitter signals and background emissions from different spatial locations. The varied projections enable the system to distinguish between signal and background by analyzing the spatial patterns detected by different elements.
Solution Approach 2:
The detector elements are deliberately configured with asymmetric or non-congruent projections into the sample, breaking the symmetry that would otherwise make background separation impossible. This asymmetric configuration ensures that each detector element experiences a unique combination of emitter and background light, providing the information needed to disentangle signal from background through computational analysis.
3Productivity
If background estimation is performed sequentially rather than simultaneously with emitter detection, then the device complexity is reduced, but the productivity decreases due to additional measurement time
Solution Approach 1:
The system performs both emitter detection and background estimation simultaneously and continuously using the same detector elements. Rather than alternating between separate measurement modes, the detector array continuously captures light emissions from both emitters and background sources in parallel, eliminating idle time and maximizing the information obtained from each measurement interval.
Solution Approach 2:
The same detector elements serve multiple functions: detecting emitter light emissions for position determination and simultaneously detecting background emissions for background correction. This multi-functionality eliminates the need for separate detection systems or sequential measurement protocols, improving productivity without adding device complexity.
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 improves the accuracy of emitter position determination by effectively addressing inhomogeneous background distributions and enabling simultaneous detection of emitter and background light, reducing measurement errors and increasing the speed of background estimation.
Implementation Method 1
The individual light-emitting emitters are, in particular, fluorophores and the illumination light is, in particular, excitation light which excites the fluorophores, whereupon they emit fluorescent light.
Data Source
AI summary
Embodiments of the invention relates to a method for localizing or tracking emitters in a sample, wherein the sample is illuminated with an intensity distribution of an illumination light having a local minimum, wherein the illumination light induces or modulates light emissions of the emitters, and wherein the local minimum is positioned in a region around a presumed position of an emitter in the sample, detecting light emissions (L) of the emitter, and determining the position of the emitter in the sample, wherein light emanating from the sample is detected with a plurality of detector elements having respective active areas whose projections into a focal plane in the sample are not congruent, wherein a background is estimated based on the light detected by the plurality of detector elements, and wherein a background correction is performed, a light microscope and a computer program for performing the method.