Single-Particle Localization Microscopy With Asymmetric Test Patterns

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Solution Overview

Problem

Existing methods for single-particle localization in light microscopy suffer from ambiguity in determining the actual particle position due to symmetry in light patterns, leading to inefficient and time-consuming solutions that involve mechanical repositioning and high photon usage.

Innovation Solution

A single-particle localization microscope that uses a sequence of symmetrical light patterns followed by an asymmetrical test pattern to unambiguously locate a particle, leveraging electro-optical or acousto-optical devices for fast pattern generation without mechanical repositioning.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If a sequence of symmetrical light patterns is used for localization, then measurement precision is improved, but ambiguity in determining actual particle position occurs

Engineering Contradiction:
Improvelocalization precisionVSAvoidposition ambiguity
Core Design Contradiction:
Measurement precisionVSLoss of information

Solution Approach 1:

The patent applies asymmetry by introducing at least one asymmetrical light pattern into the sequence of illumination patterns. This asymmetrical pattern breaks the symmetry that causes multiple potential positions to have equal likelihood, thereby resolving the position ambiguity while maintaining the high precision achieved through the sequence of measurements.

Inventive Principle:
Principle #4Asymmetry

2Measurement precision

If mechanical repositioning is used to verify particle position, then position accuracy is improved, but time consumption increases

Engineering Contradiction:
Improveposition accuracyVSAvoidmeasurement time
Core Design Contradiction:
Measurement precisionVSLoss of time

Solution Approach 1:

The patent replaces mechanical repositioning with an optical solution. Instead of physically moving the microscope or scanning device to verify particle positions, the system uses asymmetrical light patterns to optically distinguish between potential positions, eliminating mechanical movement and significantly reducing measurement time.

Inventive Principle:
Principle #28Mechanics substitution (Replace mechanical system)

3Reliability

If repeated measurement sequences are performed to resolve ambiguity, then position certainty is improved, but photon budget is depleted

Engineering Contradiction:
Improveposition certaintyVSAvoidphoton budget
Core Design Contradiction:
ReliabilityVSLoss of substance

Solution Approach 1:

The patent performs preliminary action by incorporating asymmetrical light patterns into the initial measurement sequence. This allows the system to resolve position ambiguity during the first pass of measurements, eliminating the need for repeated measurement sequences and conserving the photon budget while achieving reliable position determination.

Inventive Principle:
Principle #10Preliminary action

4Speed

If electro-optical or acousto-optical devices are used for pattern generation, then speed is improved, but device complexity increases

Engineering Contradiction:
Improvepattern generation speedVSAvoidoptical system complexity
Core Design Contradiction:
SpeedVSDevice complexity

Solution Approach 1:

The patent replaces mechanical scanning devices with electro-optical or acousto-optical devices for generating light patterns. These devices use electrical or acoustic fields to modulate light, enabling fast pattern generation without mechanical movement. While the optical system becomes more complex, the elimination of mechanical components and the speed improvement represent a favorable trade-off for dynamic localization applications.

Inventive Principle:
Principle #28Mechanics substitution (Replace mechanical system)

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

Enables fast and efficient localization of single particles with reduced photon usage by distinguishing between potential positions using an asymmetrical test pattern, eliminating the need for mechanical repositioning and improving precision.

Implementation Method 1

illuminate a sample region with a sequence of light patterns having spatially different distributions of illumination light adapted to cause a single particle located in the sample region to emit detection light

Methodology Applied
Scientific EffectFluorescence: Fluorescence

Implementation Method 2

a detector configured to detect a sequence of intensities of the detection light emerging from the sample region in response to the sequence of illuminating light patterns

Methodology Applied
Scientific EffectPhotoelectric Effect: Photoelectric Effect

Data Source

PatentEP4215970B1Single-particle localization microscope
Publication Date: 2026.04.08 LEICA MICROSYSTEMS CMS GMBH
  • EP4215970B1 patent drawingFigure 1
  • EP4215970B1 patent drawingFigure 2
  • EP4215970B1 patent drawingFigure 3~4

AI summary

A single-particle localization microscope (100) comprises an optical system (102) configured to illuminate a sample region (120) with a sequence of light patterns (124-1, 124-2, 124-3) having spatially different distributions of illumination light (110) adapted to cause a single particle (122) located in the sample region (120) to emit detection light (112), a detector (116) configured to detect a sequence of intensities (11, 12, 13) of the detection light (112) emerging from the sample region (120) in response to the sequence of illuminating light patterns (124-1, 124-2, 124-3), and a processor (132) configured to determine, based on the sequence of intensities (I1, I2, 13) of the detection light (112), an arrangement of potential positions (446a, 446b, 448a, 448b) for locating the particle (4) in the sample region (120). The processor (132) is further configured to cause the optical system (102) to illuminate the sample region (120) with at least one subsequent light pattern (524) having a spatial distribution of the illumination light (110) which is asymmetrical with respect to the arrangement of the multiple potential positions (446a, 446b, 448a, 448b, to cause the detector (116) to detect at least one subsequent intensity (14) of the detection light (112) emerging from the sample region (120) in response to the at least one subsequent light pattern (524), and to decide, based on the at least one subsequent intensity (14) of the detection light (112), which one of the multiple potential positions (446a, 446b, 448a, 448b) represents an actual position of the particle (122) in the sample region (120).