Light Microscope Refractive Index Determination via Focus Position
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Solution Overview
Problem
Light microscopes face challenges in achieving optimal image quality due to unknown or varying wavelength-dependent refractive indices of specimen media, leading to aberrations and color errors, especially in light sheet fluorescence microscopes where the refractive index of the specimen medium deviates from the ideal value.
Innovation Solution
A method and light microscope configuration that determine the wavelength-dependent refractive index of a specimen medium by measuring the focus position of illumination and detection light using a mathematical model, allowing for real-time adjustment of microscope settings to compensate for refractive index deviations.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If a specimen medium with a known wavelength-dependent refractive index is used, then image quality is improved, but costs increase
Solution Approach 1:
The light microscope automatically determines the wavelength-dependent refractive index of the specimen medium through mathematical modeling of focus positions at different wavelengths, eliminating the need for users to manually select or purchase pre-adjusted expensive media. The system self-calibrates by measuring focus positions and calculating the refractive index spectrum, making the expensive pre-adjusted media unnecessary.
2Manufacturing precision
If the refractive index of the specimen medium is adjusted before adding the specimen, then image quality is improved, but the specimen's impact on refractive index is not considered
Solution Approach 1:
The system performs focus position measurements at multiple wavelengths after the specimen is added to the medium, then uses these measurements to calculate the actual wavelength-dependent refractive index. This feedback loop allows the system to account for the specimen's impact on the refractive index and adjust the optical parameters accordingly, making the process adaptive to different specimens.
3Measurement precision
If additional apparatuses are used to determine the refractive index upfront, then measurement accuracy is improved, but device complexity and time requirements increase
Solution Approach 1:
The light microscope uses its existing illumination and detection optics to perform both specimen imaging and refractive index determination. By measuring focus positions of the illumination light at different wavelengths through the specimen medium, the system calculates the wavelength-dependent refractive index using mathematical models, eliminating the need for separate refractive index measurement devices.
4Device complexity
If the refractive index of the specimen medium deviates from the ideal value, then device simplicity is maintained, but image quality deteriorates due to aberrations
Solution Approach 1:
The system dynamically adapts to different refractive indices by measuring focus positions at multiple wavelengths and calculating the actual wavelength-dependent refractive index. Based on this measurement, the control unit automatically adjusts optical parameters such as illumination angle, detection angle, or focus position to compensate for deviations from the ideal refractive index, maintaining image quality without requiring fixed optical design.
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
Enables accurate determination of the refractive index without additional measurement units, allowing for improved image quality and reduced costs by directly determining the refractive index during specimen examination, which can include the specimen, and providing instructions for adjusting microscope settings to optimize beam paths.
Implementation Method 1
Propagation of the illumination light and the detection light in the specimen medium depends substantially on the refractive index of the specimen medium
Implementation Method 2
detection light coming from the specimen medium, wherein the detection light is in particular generated by scattering and/or fluorescence in the specimen medium
Implementation Method 3
detection light coming from the specimen medium, wherein the detection light is in particular generated by scattering and/or fluorescence in the specimen medium
Data Source
AI summary
With the method of the invention a wavelength-dependent refractive index of a specimen medium, which is examined with a light microscope, is determined. With the light microscope, a specimen measurement at the specimen medium which has an unknown refractive index is performed, wherein illumination light is radiated to the specimen medium and detection light coming from the specimen medium is measured. With the specimen measurement, a specimen measurement focus position of the illumination and/or detection light is measured. Using a mathematical model, in which a focus position of illumination and/or detection light is defined in dependence of a refractive index of a medium, the refractive index of the specimen medium is derived from the specimen measurement focus position. Furthermore, a light microscope for carrying out the method is described.