Super Resolution Microscopy via Temporal Modulation
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Solution Overview
Problem
Conventional super resolution microscopy techniques, such as SIM, are not suitable for real-time imaging of fast-moving samples due to the need for capturing multiple images at different phases and performing extensive Fourier transforms, which is time-consuming and limits the ability to observe samples in real-time with high resolution.
Innovation Solution
A sample observation device that includes an excitation light generation unit, an intermediate image forming unit, a confocal modulation unit, a modulation drive unit, an image relay unit, and an image processing unit, where the cutoff frequency of the image relay unit exceeds the optical image forming system's cutoff frequency, enabling high-frequency enhancement and real-time super resolution imaging without the need for multiple image captures.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional SIM technique captures multiple images at different phases and performs Fourier transforms, then super resolution is achieved, but imaging time increases and real-time observation becomes difficult
Solution Approach 1:
The patent extracts only the essential modulation function from the complex SIM process. By using a confocal modulation unit that directly modulates the excitation light intensity according to a temporal modulation pattern, it removes the need to capture multiple phase-shifted images and perform Fourier transforms, achieving super resolution through single-frame temporal modulation
Solution Approach 2:
The patent replaces the mechanical/optical phase-shifting system with an electronic temporal modulation system. Instead of physically shifting phases across multiple images, the confocal modulation unit applies temporal intensity modulation to the excitation light, substituting complex optical-mechanical operations with simpler electronic control
2Measurement precision
If multiple images are captured for super resolution processing, then resolution exceeds optical limit, but ability to observe fast-moving samples in real-time deteriorates
Solution Approach 1:
The patent employs periodic temporal modulation of the excitation light intensity through the confocal modulation unit. The modulation pattern varies periodically in time, encoding spatial frequency information into temporal variations that can be decoded from a single image, enabling super resolution without multiple captures
Solution Approach 2:
The patent changes the temporal parameter of excitation light intensity through periodic modulation. By varying the intensity parameter over time according to a specific pattern, the system encodes additional spatial frequency information that exceeds the optical cutoff frequency, achieving super resolution from single-frame data
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 the generation of super resolution images with high time resolution, allowing for real-time observation of samples without degrading image quality, even when the sample is moving quickly.
Implementation Method 1
an excitation light generation unit for generating excitation light for illuminating a sample S; an intermediate image forming unit for projecting the excitation light to the sample S, and forming an intermediate image of the sample S at an intermediate image position from observing light generated by illuminating the sample S with the excitation light
Data Source
Figure 1A
Figure 1B
Figure 2A
AI summary
A sample observation device (1, 20, 40, 60, 80) includes: an excitation light generation unit (2, 21, 41, 61, 81) generating excitation light for exciting a sample; an intermediate image forming unit (3, 22, 42, 62, 82) projecting the excitation light to the sample, and forming an intermediate image of the sample at an intermediate image position from observing light generated by illuminating the sample with the excitation light; a confocal modulation unit (4, 23, 44, 63, 83) modulating spatial intensity distributions of the excitation light and the intermediate image at the intermediate image position; a modulation drive unit (5, 24, 45, 64, 84) moving a modulation pattern of the confocal modulation unit relative to the intermediate image; an image relay unit (6, 25, 46, 65, 85) relaying on the image forming surface the intermediate image whose spatial intensity distribution has been modulated; an image pickup unit (7, 26, 47, 66, 86) converting the spatial intensity distribution of the intermediate image relayed on the image forming surface into digital image data; and an image processing unit (8, 27, 48, 67, 68, 87, 88) processing an image on the digital image data. The cutoff frequency of the image relay unit exceeds the cutoff frequency of the intermediate image forming unit, the Nyquist frequency of the image pickup unit exceeds the cutoff frequency of the intermediate image forming unit, and the image processing unit performs a high frequency enhancing process for enhancing the high frequency component exceeding the cutoff frequency of the intermediate image forming unit.