Light Microscopy Scanning Layout for Confocal Signal Correction
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Solution Overview
Problem
Existing light microscopy methods using tandem scanners, such as galvanometer and electro-optical scanners, suffer from signal loss due to polarization dependence and deviations from confocal detection, which affect image quality and position estimation, especially in MINFLUX techniques.
Innovation Solution
A light microscopy method utilizing two independent scanning devices, where the second scanning device is in the common illumination and detection beam path, and the first scanning device is only in the illumination path, with a detector comprising multiple elements in a detection plane, allowing emission light to be detected without descanning, and accounting for the relative position between the focus and detection plane to correct for deflections.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Speed
If emission light is scanned by an electro-optical scanner in a tandem scanner configuration, then scanning speed and positional accuracy are improved, but signal loss occurs due to polarization dependence
Solution Approach 1:
The patent divides the scanning function into two independent scanners: a first scanner (e.g., galvanometer) that scans illumination light only, and a second scanner (e.g., electro-optical) that scans both illumination and detection beam paths. This segmentation allows the electro-optical scanner to operate in a confocal arrangement where it scans both beams simultaneously, eliminating polarization-dependent signal loss while maintaining high scanning speed.
Solution Approach 2:
The patent introduces an intermediary computational step that calculates relative position between the focus and detection plane based on deflection angles from both scanners. This computational intermediary corrects for any residual misalignment, allowing the system to fully utilize the electro-optical scanner's speed advantages without suffering from polarization-related signal loss.
2Measurement precision
If a tandem scanner configuration is used with confocal detection, then measurement precision is improved, but device complexity increases
Solution Approach 1:
The patent makes the second scanner universal by using it to scan both the illumination beam path and the detection beam path. This multi-functionality simplifies the overall system architecture compared to using two separate specialized scanners, while still achieving confocal detection and high position estimation accuracy through the calculated relative position correction.
3Device complexity
If the detection plane is fixed while the focus position changes due to scanner deflection, then device simplicity is maintained, but image quality deteriorates due to deviation from confocal detection
Solution Approach 1:
The patent implements a feedback mechanism where the relative position between the focus and detection plane is continuously calculated based on real-time deflection angles from both scanners. This calculated relative position is then used to correct the detected signal, creating a feedback loop that maintains confocal detection accuracy without requiring complex mechanical adjustments to the detection plane.
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 enables the use of polarization-dependent scanners without signal loss, improving image quality and position estimation by correcting for deflections, enhancing resolution and signal-to-noise ratio in light microscopy.
Implementation Method 1
a first scanning device and a second scanning device are arranged one after the other in the illumination beam path, each configured to shift the focus of the illumination light in the sample
Implementation Method 2
a detector comprising a plurality of detector elements arranged in a detection plane that detect the emission light
Data Source
Figure 1~2
Figure 3
AI summary
The invention relates to a light microscopy method, wherein a focus of the illumination light (B) is generated in the sample (2), wherein emission light (E) emanating from emitters in the sample (2) reaches a detector (6) in a detection beam path (5), wherein the detector (5) comprises a plurality of detector elements (60) arranged in a detection plane (D) which detect the emission light (E), wherein a first scanning device (7) and a second scanning device (8) are arranged one after the other in the illumination beam path (4), each configured to shift the focus of the illumination light (B) in the sample (2), wherein the first scanning device (7) is arranged only in the illumination beam path (4), so that the emission light (E) is scanned only by the second scanning device (8).wherein a relative position (R) between a center (Z) of a projection (P) of the detection plane (D) into the sample (2) and a focus position (F) of the illumination light (B) in the sample (2) is determined, wherein detection signals acquired by the detector elements (60) are evaluated taking into account the determined relative position (R), and wherein a light microscope (1), an illumination and detection device and a computer program for carrying out the method are provided.