Microscope Sample Holder Thickness Determination
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Solution Overview
Problem
The thickness of sample holders in microscopes, such as cover glasses and Petri dish bases, can deviate from nominal values, causing aberrations that affect imaging quality, particularly at high numerical apertures, and existing methods lack a reliable way to determine and correct for these deviations in real-time.
Innovation Solution
A method involving the alignment and measurement of reflected illumination radiation to determine the thickness of sample holders, using the displacement distance between the holder's side surfaces to calculate the thickness, and adjusting correction elements like Alvarez plates or deformable mirrors to compensate for aberrations.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If the thickness of the sample holder is not precisely controlled, then manufacturing is easier and cost is lower, but imaging quality deteriorates due to aberrations
Solution Approach 1:
The patent measures the actual thickness parameter of the sample holder and uses this measured value to adjust correction elements (such as Alvarez plates or deformable mirrors) to compensate for aberrations. This transforms the manufacturing tolerance issue into a measurable and correctable parameter, allowing standard manufacturing processes to be used while maintaining high imaging quality through dynamic correction.
2Reliability
If correction elements are added to compensate for aberrations, then imaging quality is improved, but device complexity increases
Solution Approach 1:
The system automatically measures the sample holder thickness and uses this information to self-adjust the correction elements without requiring manual intervention. The measurement system and correction system are integrated, allowing the system to self-correct for aberrations based on the actual sample holder being used, thereby reducing operational complexity while maintaining imaging quality.
Solution Approach 2:
The thickness measurement and correction element adjustment are performed before actual sample imaging begins. This preliminary characterization of the sample holder allows the system to be pre-configured for optimal performance, eliminating the need for complex real-time adjustments during imaging and simplifying the overall operational process.
3Device complexity
If manual alignment and measurement methods are used, then equipment cost is lower, but measurement precision and efficiency deteriorate
Solution Approach 1:
The patent replaces manual mechanical measurement methods with an optical measurement system that uses illumination radiation and detection of reflected or transmitted light to automatically determine sample holder thickness. This substitution of mechanical measurement with optical measurement provides higher precision and automation while integrating seamlessly with the existing microscopy optical path.
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 method allows for precise determination and correction of sample holder thickness, improving image quality by reducing aberrations and enhancing the signal-to-background ratio in light sheet microscopy.
Implementation Method 1
In a step C, at least one beam of the illumination radiation is directed onto the adjusted sample holder along a first optical axis (illumination axis) at an illumination angle. In step D, at least two measured values of a reflected portion of the illumination radiation are recorded
Data Source
AI summary
The invention relates to a method for determining the thickness (d) of a sample holder (7) in the beam path of a microscope (1). The sample holder (7) is transparent to illumination radiation (BS) and is designed to receive a sample (5). The sample holder (7), which has a first side surface (OS) and a second side surface (US), is arranged in a sample plane (4), and the first side surface (OS) and second side surface (US) of the sample holder (7) are aligned parallel to the sample plane (4). At least one beam of illumination radiation (BS) is directed along a first optical axis (A1) at an illumination angle onto the aligned sample holder (7), and at least two measurements of a reflected component (BSref) of the illumination radiation (BS) or at least two measurements of a detection radiation (DS) caused by the illumination radiation (BS) are recorded.Depending on the at least two measured values, the distance between the first and second side surfaces (OS, US) in the direction of the Z-axis (Z) is determined as the thickness (d). The invention further relates to a method for acquiring image data.