Parallel Light Sheet Microscopy for High-Throughput Imaging

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Solution Overview

Problem

Light sheet microscopy faces limitations in recording speed and compatibility with standard sample holders, particularly due to the thickness of the light sheet and complex sample preparation, which restricts high-throughput screening and imaging of small samples with high resolution.

Innovation Solution

The implementation of multiple parallel light sheets that are shifted relative to each other in the detection direction, allowing for simultaneous illumination and detection of narrow strips of the sample, optimizing the use of the sensor area and enabling faster recording without compromising imaging quality or increasing radiation exposure.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If multiple parallel light sheets are used to increase recording speed, then productivity is improved, but device complexity increases

Engineering Contradiction:
Improverecording speedVSAvoidillumination device complexity
Core Design Contradiction:
ProductivityVSDevice complexity

Solution Approach 1:

The illumination is divided into multiple parallel light sheets instead of using a single light sheet. Each light sheet illuminates a separate strip of the sample, allowing simultaneous acquisition of multiple image strips. This segmentation of the illumination path enables parallel imaging, thereby increasing recording speed while distributing the complexity across multiple independent but simpler light sheet generation paths.

Inventive Principle:
Principle #1Segmentation

2Measurement precision

If the light sheet is made thinner to improve axial resolution, then measurement precision is improved, but the light sheet becomes more difficult to generate and maintain

Engineering Contradiction:
Improveaxial resolutionVSAvoidlight sheet generation complexity
Core Design Contradiction:
Measurement precisionVSDevice complexity

Solution Approach 1:

Instead of attempting to generate a single extremely thin light sheet, the system segments the illumination into multiple parallel light sheets that are slightly thicker individually but collectively provide high resolution through the parallel imaging approach. This segmentation allows each light sheet to be generated with standard optics while achieving overall high axial resolution through the multi-strip parallel acquisition.

Inventive Principle:
Principle #1Segmentation

3Ease of operation

If standard sample holders are used to simplify sample preparation, then ease of operation is improved, but the sample preparation and holder design become incompatible with traditional light sheet microscopy

Engineering Contradiction:
Improvesample preparation simplicityVSAvoidcompatibility with light sheet microscopy
Core Design Contradiction:
Ease of operationVSAdaptability or versatility

Solution Approach 1:

The illumination device is designed to generate multiple parallel light sheets that can illuminate different regions of the sample simultaneously. This multi-functionality allows the system to work with standard sample holders (multiwell plates, Petri dishes, microscope slides) that are commonly used in fluorescence microscopy, making the illumination device adaptable to various sample preparation methods and holder types while maintaining compatibility with light sheet microscopy principles.

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

This approach significantly increases the recording speed while maintaining high imaging quality and allowing the use of standard sample holders, effectively addressing the limitations of traditional light sheet microscopy in terms of sample size and preparation complexity.

Implementation Method 1

illumination optics for generating a light sheet to illuminate a strip of a sample and excite fluorescence radiation

Methodology Applied
Scientific EffectFluorescence: Fluorescence

Implementation Method 2

imaging optics for imaging the fluorescence radiation emitted by the sample onto the sensor

Methodology Applied
Scientific EffectOptical imaging: Lens

Data Source

PatentEP3304167B1Light sheet microscopy arrangement and method
Publication Date: 2021.04.14 CARL ZEISS MICROSCOPY GMBH
  • EP3304167B1 patent drawingFigure 1~2
  • EP3304167B1 patent drawingFigure 3
  • EP3304167B1 patent drawingFigure 4

AI summary

The invention relates to a light sheet microscopy arrangement comprising a lighting device (3) for generating a light sheet (LB1) in order to light a sample strip and comprising a detection device for detecting a fluorescence radiation emitted by the sample. This is achieved by a lighting device, which is designed to generate at least one additional light sheet (LB2, LB3) that is arranged parallel to a first light sheet (LB1) in order to light an additional sample strip (1), and advantageously by a detection device (4), which is designed to simultaneously detect florescence radiation excited by the light sheets (LB1, LB2, LB3) arranged parallel to one another. The invention further relates to a corresponding method.