Microscope Astigmatic Difference Adjustment for Z-Coordinate Precision
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Solution Overview
Problem
In STORM-based three-dimensional microscopy, changes in magnification or numerical aperture of the objective lens can lead to incorrect measurement of the Z-coordinate of the sample.
Innovation Solution
The microscope apparatus optimizes astigmatic difference based on image acquisition conditions by using a cylindrical lens unit with adjustable focal lengths, ensuring accurate Z-coordinate measurement by adjusting the astigmatic difference to be within an optimal range relative to the depth of focus of the objective lens.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If the magnification or numerical aperture of the objective lens is changed according to image acquisition conditions, then the adaptability to different imaging requirements is improved, but the measurement precision of the Z-coordinate deteriorates
Solution Approach 1:
The patent applies dynamics by making the astigmatic difference adjustable rather than fixed. The astigmatic difference changing device allows the astigmatic difference to be dynamically adjusted according to the depth of focus of the objective lens, enabling the system to adapt to different magnifications and numerical apertures while maintaining accurate Z-coordinate measurement. This resolves the contradiction by allowing the system to change its optical properties in response to different imaging conditions.
Solution Approach 2:
The patent changes the optical parameter (astigmatic difference) to match changes in objective lens parameters (magnification and numerical aperture). When the objective lens parameters are changed, the astigmatic difference is correspondingly adjusted to maintain the optimal relationship for Z-coordinate measurement. This ensures that measurement precision is maintained across different imaging conditions while preserving adaptability.
2Device complexity
If a fixed astigmatic difference is used in the imaging optical system, then the device complexity is reduced, but the measurement precision of the Z-coordinate deteriorates when objective lens parameters change
Solution Approach 1:
Instead of using a fixed astigmatic difference, the patent implements a dynamic adjustment mechanism that modifies the astigmatic difference based on the objective lens parameters. This dynamic approach increases device complexity slightly but ensures high measurement precision across varying imaging conditions, resolving the contradiction between simplicity and accuracy.
Solution Approach 2:
The system incorporates feedback by detecting the depth of focus of the objective lens and using this information to adjust the astigmatic difference accordingly. This feedback mechanism ensures that the astigmatic difference remains optimal for Z-coordinate measurement regardless of changes in objective lens parameters, maintaining measurement precision without requiring a completely complex system redesign.
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 high-resolution, accurate three-dimensional imaging by maintaining optimal astigmatic difference, even when magnification or numerical aperture of the objective lens changes, thereby ensuring precise Z-coordinate determination.
Implementation Method 1
the image of a fluorescent material can be made to have an elliptical shape by giving a predetermined astigmatic difference to the image of the sample by inserting a cylindrical lens in the imaging optical system
Implementation Method 2
a fluorescent material that fluoresces when irradiated with the excitation light
Data Source
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AI summary
There is provided a microscope apparatus (100) including: a plurality of objective lenses (12) having different magnifications; an imaging system (7) that receives light, which is generated from a sample (8) and emitted from the objective lens (12) when excitation light (L2) is emitted to a sample including a fluorescent material that is activated when irradiated with activation light (L1) having a predetermined wavelength and fluoresces to be inactivated when irradiated with excitation light (L2) having a different wavelength from the activation light (L1) in the activation state and that images the light in a state where an astigmatic difference is given to the image of the sample (8); and an imaging device (14) that captures the image of the sample (8) from the imaging system (7). The imaging system (7) includes an astigmatic difference changing device (50) that changes the astigmatic difference according to the depth of focus of the objective lens (12).