Point-Scanning Structured Illumination for Low-Power Super-Resolution
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Existing super-resolution fluorescence microscopy techniques face limitations such as slow imaging speed (STORM), high-power requirements (STED), and unsuitability for live cell imaging due to saturated excitation (SSIM), preventing resolution improvements beyond the diffraction limit under low-power conditions.
Innovation Solution
A point-scanning structured illumination-based system using a first laser assembly for sinusoidal excitation light and a second laser assembly for annular STED light, combined with a scanner and detector, to generate and collect fluorescence signals, and a computing terminal for image reconstruction, achieving super-resolution imaging without high power.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If STORM technique is used to achieve super-resolution imaging, then resolution exceeds diffraction limit, but imaging speed becomes slow due to requiring tens of thousands of raw images
Solution Approach 1:
The patent employs periodic structured illumination patterns that modulate fluorophore emission in a time-dependent manner. By using sinusoidal intensity modulation at specific frequencies, the system encodes spatial information into temporal frequency domains, enabling super-resolution reconstruction from fewer frames compared to STORM while maintaining imaging speed.
Solution Approach 2:
The system changes the temporal frequency parameter of excitation light intensity to encode spatial information. By modulating excitation intensity sinusoidally at different frequencies and phases, the patent transforms spatial resolution information into frequency domain parameters that can be extracted through spectral analysis, achieving super-resolution without requiring tens of thousands of images.
2Measurement precision
If STED technique is used to improve imaging resolution, then resolution exceeds diffraction limit, but high-power STED beam is required which limits the application
Solution Approach 1:
The patent changes the temporal frequency parameter of excitation light to encode spatial information in the frequency domain. By using sinusoidal modulation at specific frequencies and performing spectral analysis on the emitted fluorescence, the system achieves super-resolution without requiring high-power STED beams, thus reducing photodamage and enabling live-cell imaging applications.
Solution Approach 2:
The patent replaces the mechanical/optical approach of STED (using high-power depletion beams to suppress fluorescence) with a frequency-domain encoding approach. Instead of physically suppressing fluorescence with high-power beams, the system uses temporal frequency modulation and spectral analysis to achieve super-resolution, substituting a less invasive optical mechanism.
3Power
If SIM technique is used to improve imaging resolution, then imaging can be performed with common fluorophores and low excitation light intensity, but resolution can only be improved by two times due to diffraction limit
Solution Approach 1:
The patent uses periodic structured illumination with sinusoidal intensity modulation to encode spatial information into temporal frequency domains. By analyzing the frequency spectrum of the emitted fluorescence signal, the system achieves super-resolution beyond the conventional two-fold improvement of SIM, while maintaining low excitation light intensity suitable for live-cell imaging.
Solution Approach 2:
The patent introduces temporal frequency modulation as an additional parameter dimension beyond spatial patterning. By modulating excitation intensity sinusoidally at specific frequencies and extracting frequency components from the emission signal, the system achieves higher resolution than conventional SIM without increasing excitation power.
4Measurement precision
If SSIM technique is used to achieve super-resolution imaging, then resolution exceeds diffraction limit, but extremely high optical power is required for saturated excitation which is not applicable to live cell imaging
Solution Approach 1:
The patent employs periodic sinusoidal modulation of excitation light intensity at specific frequencies to encode spatial information temporally. This approach achieves super-resolution without requiring saturated excitation conditions, thereby avoiding the need for extremely high optical power and making the technique applicable to live-cell imaging where photodamage must be minimized.
Solution Approach 2:
The patent changes from spatial-only modulation (conventional SIM/SSIM) to temporal frequency modulation. By sinusoidally varying excitation intensity at specific frequencies and analyzing the frequency spectrum of emission, the system achieves super-resolution without saturated excitation, thus operating at low optical power suitable for live cells.
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 system achieves super-resolution imaging up to tens of nanometers with improved imaging speed and suitability for live cell imaging by using sinusoidal excitation and annular STED light, doubling the resolution compared to conventional methods.
Implementation Method 1
the first laser assembly is configured to generate an excitation light whose intensity varies sinusoidally with time
Implementation Method 2
the second laser assembly is configured to generate an annular stimulated emission depletion (STED) light
Implementation Method 3
the scanner is configured to control the excitation light and the annular STED light to scan and excite a to-be-imaged sample to generate fluorescence signals
Implementation Method 4
the detector is configured to acquire the fluorescence signals to obtain a plurality of fluorescence-structured images in different orientations and phases
Implementation Method 5
the computing terminal is configured to extract frequency components of each of the plurality of fluorescence-structured images, and reconstruct a plurality of local super-resolution images
Data Source
AI summary
A point-scanning structured illumination-based super-resolution microscopic imaging system includes a first laser assembly, a second laser assembly, a scanner, a detector and a computing terminal. The first laser assembly and the second laser assembly are configured to generate an excitation light and an annular STED light, respectively. The scanner is configured to control the excitation light and the annular STED light to scan and excite a sample. The detector is configured to acquire fluorescence signals to obtain fluorescence-structured images. The computing terminal is configured to reconstruct a super-resolution image based on the fluorescence-structured images. A microscopic imaging method is also provided, in which the to-be-imaged sample is scanned and excited by an excitation light and an annular STED light to obtain a stripe structured-light image beyond the diffraction limit, and the fluorescence signals are collected by a detector synchronously and pointwise in real time.