Light Sheet Microscope Beam Modulation for Resolution

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Solution Overview

Problem

Conventional light sheet microscopy using Gaussian beams is limited by resolution in the detection direction, scaling issues with image field size, limited depth of penetration into scattering media, and uniform illumination, particularly in OPM and SCAPE applications.

Innovation Solution

The use of beam modulation means, such as beam shaping optics that generate Bessel or Mathieu beams, which redistribute illumination light to achieve better resolution and penetration depth, and include a diaphragm arrangement or spatial light modulators to modulate the illumination beam along the illumination axis, compensating for scattering and absorption effects.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If a Gaussian beam is used for illumination, then the illumination is uniform and easy to generate, but the resolution in the detection direction is limited and the depth of penetration into scattering media is limited

Engineering Contradiction:
Improveresolution in detection directionVSAvoidbeam modulation complexity
Core Design Contradiction:
Measurement precisionVSDevice complexity

Solution Approach 1:

The patent changes the beam profile parameter from Gaussian to Bessel or Mathieu beams through beam shaping optics. This parameter change enables self-reconstructing properties that improve resolution in the detection direction by maintaining beam intensity through scattering media, while the beam modulation means (spatial light modulator or beam shaping optics) provides the necessary control without excessive complexity

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent introduces beam shaping optics or spatial light modulators as intermediary elements between the light source and sample. These intermediaries transform the Gaussian beam into Bessel or Mathieu beams with self-reconstructing properties, enabling improved penetration depth and resolution without directly modifying the light source itself

Inventive Principle:
Principle #24Intermediary (Mediator)

2Device complexity

If the light sheet is tilted for OPM or SCAPE applications, then single lens can be used for both illumination and detection, but spherical aberration and scattering effects increase

Engineering Contradiction:
Improveoptical system simplificationVSAvoidspherical aberration and scattering
Core Design Contradiction:
Device complexityVSObject-affected harmful factors

Solution Approach 1:

The patent changes the illumination beam profile parameter to Bessel or Mathieu beams, which have self-reconstructing properties that counteract spherical aberration and scattering effects. This allows the tilted light sheet configuration to maintain image quality despite the increased harmful factors, while keeping the optical system simplified with a single lens

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent utilizes the self-reconstructing property of Bessel and Mathieu beams to convert the harmful effects of scattering and spherical aberration into beneficial outcomes. The beams automatically reconstruct their profile after being disturbed by scattering media or spherical aberration, turning these detrimental factors into opportunities to demonstrate the robustness and superior performance of the proposed illumination method

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

3Measurement precision

If beam modulation means are introduced to improve resolution and penetration depth, then image quality improves, but the device complexity and manufacturing difficulty increase

Engineering Contradiction:
Improveimage resolution and contrastVSAvoidmanufacturing and alignment difficulty
Core Design Contradiction:
Measurement precisionVSEase of manufacture

Solution Approach 1:

The patent employs beam shaping optics or spatial light modulators as intermediary components that can be integrated into existing light sheet microscopy systems. These intermediaries provide the necessary beam modulation functionality without requiring complete system redesign, thereby improving manufacturability while achieving enhanced image resolution and penetration depth

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 enhances resolution in the detection direction, improves penetration depth, and maintains image contrast by self-reconstructing beams and adaptive intensity modulation, reducing spherical aberration and scattering effects.

Implementation Method 1

beam shaping optics that generate Bessel or Mathieu beams

Methodology Applied
Scientific EffectBessel beam generation:

Implementation Method 2

beam shaping optics that generate Bessel or Mathieu beams

Methodology Applied
Scientific EffectMathieu beam generation:

Implementation Method 3

compensating for scattering and absorption effects

Methodology Applied
Scientific EffectScattering compensation: Scattering

Implementation Method 4

compensating for scattering and absorption effects

Methodology Applied
Scientific EffectAbsorption compensation: Absorption (EM radiation)

Implementation Method 5

a beam source (14a, 14c, 14d) which is designed to direct an illumination beam (16) onto a sample (30)

Methodology Applied
Scientific EffectFluorescence excitation: Fluorescence

Data Source

PatentEP3497501B1Light sheet microscope
Publication Date: 2022.01.19 LEICA MICROSYSTEMS CMS GMBH
  • EP3497501B1 patent drawingFigure 1~2
  • EP3497501B1 patent drawingFigure 3~4
  • EP3497501B1 patent drawingFigure 5~6

AI summary

The invention relates to a light sheet microscope (10a, 10b, 10c, 10d) comprising an illuminating unit (12) having a beam source (14a, 14b, 14c, 14d), which is designed to direct an illuminating beam (16) propagating along an illumination axis (z) toward a sample (30), a light-sheet producing means (20b, 62, 72), which is designed to produce a light-sheet-type illuminating light distribution that illuminates the sample (32) in a partial region from the illuminating beam (16), a detection unit (39), which has a detector (44a, 44b, 44c, 44d), which is designed to sense detection light (34), which originates from the partial region of the sample (30) illuminated with the illuminating light distribution, and an objective (28), which is provided jointly for the illuminating unit (12) and the detection unit (39) and through which the illuminating beam (16) and the detection light (34) pass. The illuminating unit (12) has a beam-modulating means (56, 75, 78, 110), which is designed to modulate the illuminating beam (16) perpendicularly to the illumination axis (z).