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
Engineering 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
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
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
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
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
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
3Productivity
If SPIM technology is used for rapid sample examination, then examination speed increases, but device complexity increases due to separate optical beam paths
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
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
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
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
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
Implementation Method 2
the light stripe focused by the illumination objective
Implementation Method 3
The deflection means serves to deflect a light stripe emerging from an illumination lens
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
Data Source
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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.