Microscope Polarization Modulation for Resolution
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Solution Overview
Problem
Current methods for high-resolution optical analysis of fluorescent molecules, such as STED microscopy and photo-activated localization microscopy, require precise alignment and are limited by the need for specific fluorescent molecules that can be excited by a switching wavelength, making them time-consuming and inefficient for detecting a large number of fluorescent molecules.
Innovation Solution
A device with a polarization modulator that synchronizes the excitation light's polarization with the transition dipole moment vectors of fluorescent molecules, using a modulation signal to selectively excite and detect fluorescent molecules, allowing for higher spatial resolution without the need for a switching wavelength, and optionally incorporating a de-excitation light source to further enhance resolution.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If STED microscopy with two parallel coupled light paths is used, then high resolution is achieved, but precise alignment and time-consuming scanning are required
Solution Approach 1:
The patent combines the excitation light path and de-excitation light path into a single common beam path, eliminating the need for precise alignment between separate paths. The polarization modulator is integrated into this unified path, simplifying the overall device structure while maintaining high resolution capabilities.
Solution Approach 2:
The single objective lens serves multiple functions: it focuses both the excitation light and the de-excitation light, and also collects the emitted fluorescence. This multi-functional design eliminates the need for separate optical paths and reduces alignment complexity.
2Measurement precision
If photo-activated localization microscopy with switching wavelength irradiation is used, then individual molecule detection is achieved, but the method is limited to specific fluorescent molecules and is time-consuming
Solution Approach 1:
The patent employs periodic modulation of the excitation light polarization state to selectively excite fluorescent molecules with different transition dipole moment orientations. This periodic action allows simultaneous detection of multiple molecules without requiring sequential switching, thereby increasing detection speed while maintaining high resolution.
Solution Approach 2:
The patent changes the polarization parameter of the excitation light dynamically to selectively excite different subsets of fluorescent molecules. By modulating the polarization angle and detecting the corresponding emission, the system can rapidly analyze multiple molecules simultaneously without being limited to specific fluorescent protein types.
3Adaptability or versatility
If polarization modulation with synchronization is used, then broader range of fluorescent molecules can be detected, but setup effort is required
Solution Approach 1:
The system automatically determines the orientation of transition dipole moments by analyzing the polarization-modulated emission signals. This self-calibration capability eliminates the need for manual setup effort to characterize each fluorescent molecule, allowing the system to adapt to a broad range of molecules without extensive preparation.
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
The method achieves higher spatial resolution and efficiency in detecting fluorescent molecules, reducing the effort required for setup and enabling the analysis of a broader range of fluorescent molecules, with improved detection of individual molecules and enhanced resolution in microscopic representations.
Implementation Method 1
the polarization of the excitation light is modulated with a modulation signal
Implementation Method 2
excitation light generated by an excitation light source is directed onto a sample and light emitted by the sample is detected
Data Source
Figure 1~2
Figure 3A~4F
Figure 5A~6C
AI summary
The present invention relates to a device for the optical analysis of a sample, also referred to as a microscope, which is provided for a high resolution optical analysis method for detecting fluorescent molecules. The device and the method carried out using the device are designed such that excitation light generated by an excitation light source is directed onto a sample and light emitted by the sample is detected. The device and the method are characterised in that the excitation light is synchronised with the detection process. The device is characterised by having a polarisation device provided for modulating the polarisation of the excitation light by means of a modulation signal, the modulation signal having or consisting of at least one frequency, in particular a pre-determined frequency or a plurality of pre-determined superimposed frequencies, or the modulation signal consisting of a sequence of non-repeated signals.