SPIM Microscope Light Sheet Focus Control via Beam Expanding Telescope
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Solution Overview
Problem
Current SPIM microscopes face challenges such as complex adjustments for image field size variation, light sheet alignment, and sample handling issues, which limit user-friendliness and throughput in commercial applications.
Innovation Solution
The microscope incorporates a dichroic beam splitter arranged near infinity with a thickness of at least 3 mm, wobble plates for beam offset adjustment, and a detection zoom element for flexible image field size and focus control, allowing for automated image overlay and reduced interactions with the sample chamber.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If the detection objective is changed to vary the image field size, then the field of view can be adjusted, but the sample chamber must be accessed and refocusing is required, reducing throughput
Solution Approach 1:
A beam expanding telescope is introduced as an intermediary optical element in the illumination beam path. This telescope can be adjusted to change the image field size without requiring access to the sample chamber or refocusing operations, thereby maintaining high sample throughput while providing field of view adjustment capability.
2Measurement precision
If the detection objective is moved or adjusted, then focus can be changed, but vibrations are transmitted to the sample through the immersion liquid, degrading image quality
Solution Approach 1:
The beam expanding telescope serves as a mediator that enables focus adjustment indirectly through optical path modification rather than direct movement of the detection objective. This eliminates mechanical vibrations that would otherwise be transmitted through the immersion liquid to the sample, preserving image quality while maintaining focus control capability.
3Volume of moving object
If a thin dichroic beam splitter is used, then the device is more compact, but image artifacts such as defocusing and astigmatism occur
Solution Approach 1:
The thickness parameter of the dichroic beam splitter is increased from thin to at least 3 mm. This parameter change eliminates image artifacts such as defocusing and astigmatism that occur with thin beam splitters, while the beam expanding telescope compensates for any increase in overall device volume by providing optical path adjustment capability.
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 configuration enhances the microscope's adaptability to various experimental conditions, improves image quality, and simplifies sample handling, increasing user convenience and throughput by enabling precise and efficient adjustment of image fields and focus without disturbing the sample.
Implementation Method 1
splitting means for splitting the detection beam path into two partial beam paths, wherein in each of the partial beam paths a spatially resolving area detector onto which the light to be detected is imaged is arranged, and wherein the splitting means in turn comprise at least one dichroic beam splitter
Implementation Method 2
at least one wobble plate is arranged in at least one of the two partial beam paths to generate a beam offset along two mutually orthogonal directions transverse to the detection axis. In this way, automated superpositions of the measured values read from the two area detectors can be carried out
Implementation Method 3
optical imaging elements for imaging the light to be detected onto the respective area detector are arranged in each of the two partial beam paths
Data Source
Figure 1
Figure 2
Figure 3a~3b
AI summary
The invention relates to a microscope comprising an illumination device (19) with which a light sheet for illuminating a sample area (P) is generated, which is extended approximately planarly in the direction of an illumination axis (X) of an illumination beam path and in the direction of a transverse axis (Y) which lies transverse to the illumination axis (X), further comprising a detection device (1) with which light is detected which is emitted from the sample area (P) along a detection axis (Z) of a detection beam path, wherein the illumination axis (X) and detection axis (Z) as well as the transverse axis (Y) and detection axis (Z) are at a non-zero angle to each other, and wherein the detection device further comprises a detection objective (2) in the detection beam path.In such a microscope, the illumination device (19) in the illumination beam path comprises at least one illumination objective (20) and an optical illumination element spatially separated from a front lens of the illumination objective (20) and adjustable independently of it, by means of which an illumination focus plane in the sample area (P) and thus a waist of the light sheet along the illumination axis (X) can be continuously moved.