SPIM Microscope Light Sheet Deflection for High-Throughput Imaging

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Solution Overview

Problem

Current methods for microscopic examination of large numbers of samples are inefficient, particularly for light-sensitive samples, as they require lengthy scanning times and complex setups, leading to sample damage and unsuitable for automated mass examinations.

Innovation Solution

The use of Single Plane Illumination Microscopy (SPIM) technology in conjunction with a sample holder that allows for automated and precise positioning of samples within a sample illumination position, using a sample holder with sub-holders and deflection means to facilitate efficient and gentle illumination, enabling high-throughput examination with low phototoxicity.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If conventional scanning microscopy is used to examine multiple samples, then measurement precision is maintained, but examination time increases significantly and samples are damaged by prolonged light exposure

Engineering Contradiction:
Improvemicroscopic examination qualityVSAvoidexamination time
Core Design Contradiction:
Measurement precisionVSLoss of time

Solution Approach 1:

The invention segments the illumination and detection processes by using separate optical paths. The illumination objective illuminates the sample from one direction while the detection objective captures light from a perpendicular direction, allowing parallel processing of illumination and detection functions that reduces total examination time while maintaining precision

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

The invention transitions from conventional single-plane scanning to three-dimensional examination by illuminating and detecting multiple focal planes simultaneously. The illumination objective creates light sheets at different depths while the detection objective captures images from multiple planes, enabling volumetric examination without sequential scanning

Inventive Principle:
Principle #17Another dimension (Dimensionality change)

2Measurement precision

If conventional scanning microscopy is used to examine multiple samples, then measurement precision is maintained, but sample damage increases due to prolonged light exposure

Engineering Contradiction:
Improvemicroscopic examination qualityVSAvoidphototoxicity
Core Design Contradiction:
Measurement precisionVSObject-affected harmful factors

Solution Approach 1:

By separating illumination and detection into different optical paths and temporal sequences, the system illuminates only the specific focal plane being detected at each moment, minimizing total light exposure to the sample while maintaining detection precision through the perpendicular observation angle

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

The system continuously illuminates and detects multiple focal planes in rapid succession, creating the perception of simultaneous multi-plane imaging. This continuous process reduces the total time samples are exposed to light compared to sequential scanning, thereby reducing phototoxicity while maintaining measurement precision

Inventive Principle:
Principle #20Continuity of useful action

3Productivity

If SPIM technology is used for rapid sample examination, then examination speed increases, but device complexity increases due to separate optical beam paths

Engineering Contradiction:
Improveexamination throughputVSAvoidoptical system complexity
Core Design Contradiction:
ProductivityVSDevice complexity

Solution Approach 1:

The invention makes a single sample holder and stage system serve multiple functions by enabling it to accommodate both conventional single-objective configurations and SPIM dual-objective configurations. The same mechanical infrastructure supports different examination modes, reducing overall system complexity while maintaining high throughput capability

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

Solution Approach 2:

The system dynamically adapts its optical configuration based on examination requirements. The sample holder and stage can be positioned and configured for either conventional scanning or SPIM multi-plane imaging, allowing the system to optimize between speed and complexity depending on the specific examination task

Inventive Principle:
Principle #15Dynamics

4Measurement precision

If individual sample setup is required for SPIM examination, then measurement precision is maintained, but ease of operation decreases due to laborious setup procedures

Engineering Contradiction:
Improvemicroscopic examination qualityVSAvoidsetup ease
Core Design Contradiction:
Measurement precisionVSEase of operation

Solution Approach 1:

The sample holder is designed as a universal platform that can accommodate multiple samples in a standardized array configuration. This universal design allows the same holder to be used for both conventional and SPIM examinations, eliminating the need for special individual setup procedures while maintaining the precision required for SPIM imaging through proper sample positioning

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 allows for rapid, precise, and gentle microscopic examination of light-sensitive samples, such as tissue cultures and embryonic development, with high image contrast and minimal sample loss, enabling continuous and automated mass examination without the need for complex setups.

Implementation Method 1

a layer of the sample located in the illumination position is illuminated with the light stripe focused by the illumination objective

Methodology Applied
Scientific EffectLight stripe illumination: Light

Implementation Method 2

the light stripe focused by the illumination objective

Methodology Applied
Scientific EffectOptical focusing: Focusing

Implementation Method 3

The deflection means serves to deflect a light stripe emerging from an illumination lens

Methodology Applied
Scientific EffectLight deflection: Reflection

Implementation Method 4

The sample holder, which holds the samples, is preferably mounted by motor and/or automatically so that it can be moved relative to the sample illumination position

Methodology Applied
Scientific EffectMechanical displacement: Displacement

Data Source

PatentEP3011379B1Method and optical device for microscopically examining a multiplicity of specimens
Publication Date: 2021.08.18 LEICA MICROSYSTEMS CMS GMBH
  • EP3011379B1 patent drawingFigure 1
  • EP3011379B1 patent drawingFigure 2
  • EP3011379B1 patent drawingFigure 3

AI summary

The invention relates to a method for microscopically examining a multiplicity of specimens. The method comprises the step of arranging the specimens in a specimen holder, which can be moved, in particular in a motorized and/or automatic manner, in relation to a specimen-illuminating position in such a way that at least one of the specimens at a time can be successively positioned in the specimen-illuminating position, wherein a free space for a deflecting means remains in each case adjacent to the specimen that is in the specimen-illuminating position at the time, the step of focusing a strip of light by an illumination lens, the step of deflecting the strip of light after it has passed through the illumination lens by the deflecting means in such a way that the strip of light spreads out at an angle other than zero degrees in relation to the optical axis of the illumination lens and has a focal point in the specimen-illuminating position, and the step of successively positioning the specimens secured by the specimen holder in the specimen-illuminating position and detecting the detection light that emanates from the specimen respectively located in the specimen-illuminating position. The invention also relates to an optical device with a specimen holder, which holds a multiplicity of specimens and which is mounted in such a way that it can be moved, in particular in a motorized and/or automatic manner, in relation to a specimen-illuminating position in such a way that at least one of the specimens at a time can be successively positioned in the specimen-illuminating position.