Tri-Spot PSF Phase Mask for Single-Molecule Orientation

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Solution Overview

Problem

Existing methods for single-molecule imaging struggle to measure the orientations and rotational mobilities of single-molecule emitters with high precision, particularly in 3D, due to limitations in sensitivity and complexity of optical instruments.

Innovation Solution

A tri-spot point spread function (PSF) imaging system using a custom-designed phase mask that produces a triangular pattern of three light spots, allowing for the estimation of molecular orientation and rotational mobility by analyzing the relative brightness of these spots, which is sensitive to all five degrees of freedom related to molecular orientation and mobility.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If existing single-molecule imaging methods are used to measure molecular orientation and rotational mobility, then some measurement capability is achieved, but measurement precision and sensitivity are insufficient

Engineering Contradiction:

Improvemeasurement precision of molecular orientation and rotational mobilityVSAvoidsensitivity of measurement

Core Design Contradiction:

Measurement precisionVSReliability

Solution Approach 1:

The imaging system segments the point spread function into three distinct spots arranged in a triangular pattern. Each spot's intensity independently encodes information about different aspects of molecular orientation and rotational mobility, enabling precise measurement of all five degrees of freedom simultaneously with high sensitivity

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

The patent employs an asymmetric phase mask with three distinct partitions that create an asymmetric tri-spot PSF pattern. This asymmetry ensures that each spot responds differently to molecular orientation changes, providing unique sensitivity to all five degrees of freedom (three orientation angles and two rotational mobility parameters) without requiring complex optical instruments

Inventive Principle:

Principle #4Asymmetry

2Measurement precision

If complicated optical instruments are used to measure molecular orientation and rotational mobility, then measurement capability is improved, but device complexity increases

Engineering Contradiction:

Improvecapability to measure molecular orientation and rotational mobilityVSAvoidcomplexity of optical instruments

Core Design Contradiction:

Measurement precisionVSDevice complexity

Solution Approach 1:

The tri-spot PSF imaging system performs multiple measurement functions simultaneously using a single optical configuration. It measures all five degrees of freedom (molecular orientation angles and rotational mobility parameters) in one imaging experiment, eliminating the need for multiple specialized instruments or complex tuning mechanisms

Inventive Principle:

Principle #6Universality (Multi-functionality)

Solution Approach 2:

The patent changes the optical parameter by introducing a phase mask that transforms the standard PSF into a tri-spot pattern. This parameter change in the optical system enables simultaneous measurement of multiple molecular parameters without requiring complicated optical instruments or multiple measurement setups

Inventive Principle:

Principle #35Parameter changes

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 precise measurement of molecular orientation and rotational mobility, enhancing the sensitivity and precision of single-molecule imaging beyond the diffraction limit, suitable for applications in biological research such as DNA bending and transbilayer lipid motion analysis.

Implementation Method 1

Each partition includes a phase delay ramp aligned along a phase delay axis. Each phase delay ramp includes a gradient of phase delays.

Methodology Applied

Scientific EffectPhase delay: Phase Modulation

Implementation Method 2

a phase mask positioned between the at least one emitter and the at least one sensor, the phase mask configured to produce a tri-spot point spread function in response to photons received from the at least one emitter

Methodology Applied

Scientific EffectDiffraction: Diffraction

Data Source

PatentUS10761419B2Systems and methods for performing optical imaging using a tri-spot point spread function (PSF)

Publication Date: 2020.09.01 WASHINGTON UNIV IN SAINT LOUIS

AI summary

Systems and methods for use in tri-spot point spread function imaging are provided that include a phase mask. The phase mask includes three partitions that each include a subset of the total area that is asymmetrical to other partitions. Each partition includes a phase delay ramp aligned along a phase delay axis, and includes a gradient of phase delays. Each phase delay axis is oriented in a different direction with respect to each other. Included is a source that outputs an excitation beam into a sample containing at least one light emitter that emits a radiation pattern when illuminated. Included is at least one sensor arranged to capture at least one image of the radiation pattern and a phase mask positioned between the at least one emitter and the at least one sensor. The phase mask is configured to produce a tri-spot point spread function.