Adaptive Optics Microscopy Wavefront Correction

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Solution Overview

Problem

Conventional light microscopy struggles to achieve diffraction-limited resolution in biological samples due to sample-induced aberrations, leading to image degradation and loss of signal with increasing depth, as the optical properties of biological tissues deviate from the designed immersion media, causing optical distortions and reduced image fidelity.

Innovation Solution

The technique involves individually controlling the directions and phases of beamlets at the rear pupil of an optical system using a wavefront modulating element, such as a spatial light modulator, to ensure constructive interference and adaptive optical correction for aberrations, allowing for precise focusing and enhanced signal emission.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If conventional light microscopy is used to image biological samples, then the system structure remains simple, but image resolution and signal intensity deteriorate with increasing depth due to sample-induced aberrations

Engineering Contradiction:
Improveimage resolutionVSAvoidoptical system complexity
Core Design Contradiction:
Measurement precisionVSDevice complexity

Solution Approach 1:

The patent segments the wavefront into multiple beamlets that can be independently controlled. The spatial light modulator divides the incoming wavefront into discrete regions, each corresponding to a beamlet that can be individually manipulated in direction and phase to correct aberrations at different spatial locations

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

The patent implements dynamic control of beamlet directions and phases through a spatial light modulator that can be programmed in real-time. This allows the optical system to adapt to varying aberration conditions at different depths and locations within the biological sample, transforming a static optical system into a dynamically adjustable one

Inventive Principle:
Principle #15Dynamics

2Reliability

If the optical properties of biological samples differ from immersion media, then the system design remains straightforward, but optical distortions and aberrations increase, leading to signal loss

Engineering Contradiction:
Improvesignal intensityVSAvoidoptical aberrations
Core Design Contradiction:
ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The patent converts the harmful effect of sample-induced aberrations into useful information by using the emitted light itself to probe the aberration conditions. By analyzing how light propagates through the sample and returns, the system determines the actual optical path variations and uses this information to calculate corrective phase adjustments for subsequent imaging

Inventive Principle:
Principle #22Blessing in disguise (Convert harm into benefit)

Solution Approach 2:

The patent implements a feedback mechanism where the emission light from the focal spot is detected and used to inform adjustments to the excitation beamlet phases and directions. The spatial light modulator is programmed based on detected signal characteristics to optimize focusing conditions, creating a closed-loop system that continuously adapts to sample conditions

Inventive Principle:
Principle #23Feedback

3Measurement precision

If beamlets are individually controlled to correct aberrations, then image fidelity improves, but the complexity of beam control increases

Engineering Contradiction:
Improveimage fidelityVSAvoidbeam control complexity
Core Design Contradiction:
Measurement precisionVSDevice complexity

Solution Approach 1:

The patent introduces a spatial light modulator as an intermediary device between the light source and the sample. This intermediary component performs the complex task of individual beamlet control by modulating the phase and direction of each beamlet independently, shielding the rest of the system from the complexity of direct beam manipulation while enabling precise aberration correction

Inventive Principle:
Principle #24Intermediary (Mediator)

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 effectively corrects for system and sample-induced aberrations, improving image resolution and signal intensity by optimizing the focal spot's spatial extent and phase alignment, thereby enhancing the overall imaging performance beyond conventional limits.

Implementation Method 1

directions and/or relative phases of the individual beamlets of the excitation beam at a rear pupil of the lens are individually varied with a wavefront modulating element

Methodology Applied
Scientific EffectWavefront modulation:

Implementation Method 2

the light that impinges on the rear pupil is deliberately not a plane wave but rather a wave distorted such that, after the beamlets traverse the optical system and a sample, they interfere constructively and overlap densely within the sample

Methodology Applied
Scientific EffectInterference: Interference

Implementation Method 3

focusing a beam of excitation light with a lens to a focal spot within a sample

Methodology Applied
Scientific EffectRefraction: Refraction

Implementation Method 4

a global phrase ramp can be applied to light reflected from an active layer of the spatial light modulator to induce a non-zero direction between light reflected from the active layer and light reflected from other interfaces of the spatial light modulator

Methodology Applied
Scientific EffectPhase modulation: Phase Modulation

Data Source

PatentEP2732326B1Microscopy with adaptive optics
Publication Date: 2020.11.18 HOWARD HUGHES MEDICAL INST
  • EP2732326B1 patent drawingFigure 1
  • EP2732326B1 patent drawingFigure 2
  • EP2732326B1 patent drawingFigure 3a~3c

AI summary

A method of manipulating a focused light beam includes focusing a beam of excitation light with a lens to a focal spot within a sample, where a cross-section of the beam includes individual beamlets. Directions and/or relative phases of the individual beamlets of the excitation beam at a rear pupil of the lens are individually varied with a wavefront modulating element, and emission light emitted from the focal spot is detected while the directions or relative phases of individual beamlets are varied. The directions of individual beamlets are controlled to either maximize or minimize the emission light from the focal spot, and the relative phases of individual beamlets are controlled to increase the emission light from the focal spot.