Adaptive Optics Wavefront Correction in Laser Scanning Microscopy

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Solution Overview

Problem

Conventional laser scanning microscopes face challenges in achieving high-resolution imaging due to diffraction limits and sample-induced wavefront errors, which require additional photon usage for aberration correction and are inefficient in terms of photon usage and scanning speed.

Innovation Solution

A super-resolution laser scanning microscope (SR-LSM) that includes an adaptive optics system for wavefront correction, utilizing a detector array to evaluate the point spread function (PSF) at each scanning position, allowing for location-dependent aberration correction without sacrificing photons, and enabling faster convergence of the algorithm by using previous scanning position data.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Manufacturing precision

If adaptive optics is used to correct wavefront errors, then imaging resolution is improved, but additional photons are consumed for aberration measurement and correction

Engineering Contradiction:
Improveimaging resolutionVSAvoidphoton consumption
Core Design Contradiction:
Manufacturing precisionVSQuantity of substance

Solution Approach 1:

The patent combines the aberration measurement and imaging functions into a single detection process. The same detector that captures imaging photons also measures the point spread function (PSF) to determine wavefront errors, eliminating the need for separate measurement photons. This merging allows simultaneous correction of aberrations while maintaining imaging efficiency.

Inventive Principle:
Principle #5Merging (Combining)

Solution Approach 2:

The system uses the imaging photons themselves to serve dual purposes: both for forming the image and for characterizing the PSF to enable aberration correction. The imaging data automatically provides information about wavefront errors through PSF analysis, allowing the system to self-correct without external measurement systems consuming additional photons.

Inventive Principle:
Principle #25Self-service

2Manufacturing precision

If conventional aberration correction methods are used, then wavefront errors are corrected, but scanning speed is reduced due to iterative measurement requirements

Engineering Contradiction:
Improveaberration correction accuracyVSAvoidscanning speed
Core Design Contradiction:
Manufacturing precisionVSSpeed

Solution Approach 1:

The patent enables continuous aberration correction throughout the scanning process by measuring the PSF at each scanning position and applying corrections in real-time. This continuous feedback loop maintains optimal imaging conditions without interrupting the scanning flow, unlike conventional methods that require separate measurement cycles.

Inventive Principle:
Principle #20Continuity of useful action

Solution Approach 2:

The system performs preliminary PSF measurement and aberration determination at each scanning position before final image formation. By preparing the correction data in advance during the scanning process itself, the system eliminates post-processing delays and maintains high scanning speed.

Inventive Principle:
Principle #10Preliminary action

3Measurement precision

If location-dependent aberration correction is implemented, then imaging accuracy across the field of view is improved, but system complexity increases

Engineering Contradiction:
Improveimaging accuracyVSAvoidsystem complexity
Core Design Contradiction:
Measurement precisionVSDevice complexity

Solution Approach 1:

The patent implements location-dependent aberration correction by measuring and correcting the PSF at each scanning position independently. This local approach tailors the correction to the specific conditions at each position in the field of view, improving imaging accuracy without requiring a complete system redesign.

Inventive Principle:
Principle #3Local quality

Solution Approach 2:

The system uses a universal PSF measurement and analysis approach that can be applied at any scanning position. The same detection and processing methods work throughout the entire field of view, allowing location-dependent correction without multiplying the number of different system components or procedures.

Inventive Principle:
Principle #6Universality (Multi-functionality)

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

The SR-LSM achieves improved resolution and signal-to-noise ratio by correcting wavefront errors on-the-fly or through post-processing, reducing the need for additional photon measurements and enhancing scanning speed by iteratively refining wavefront corrections.

Implementation Method 1

utilizing a detector array to evaluate the point spread function (PSF) at each scanning position, allowing for location-dependent aberration correction

Methodology Applied
Scientific EffectWavefront modulation:

Implementation Method 2

A super-resolution laser scanning microscope (SR-LSM) that includes an adaptive optics system for wavefront correction

Methodology Applied
Scientific EffectAdaptive optics:

Implementation Method 3

a laser is used for the illumination, which illuminates an object in the focal plane and excites fluorescence molecules at every point

Methodology Applied
Scientific EffectFluorescence excitation: Fluorescence

Implementation Method 4

a detector for detecting one emitted by the sample spatially resolved imaging spots

Methodology Applied
Scientific EffectLight detection: Photoelectric Effect

Data Source

PatentEP3047325B1Laser scanning microscope and method for correcting imaging aberrations in laser scanning microscopy
Publication Date: 2020.03.18 CARL ZEISS MICROSCOPY GMBH
  • EP3047325B1 patent drawingFigure 1
  • EP3047325B1 patent drawingFigure 2~3
  • EP3047325B1 patent drawingFigure 4(A)~5b

AI summary

A laser scanning microscope (SR-LSM) and a method for correcting imaging errors in a laser scanning microscope. The SR-LSM includes an illumination device for providing an illumination spot; a scanner for moving the illumination spot to consecutive scanning positions over a sample to be examined; an adaptive optics unit for controlling a wavefront of the illumination spot with a control device and a detector for determining a spatially resolved imaging spot emitted by the sample. An evaluation unit is provided for determining a point-spread function (PSF) of the imaging spot at each scanning position, whereby a wavefront correction signal determined from the point-spread function (PSF) of a scanning position is supplied to the control device of the adaptive optics unit or is used in digital post-processing of the microscope image (e.g. by means of deconvolution).