Luminescence Microscopy Resolution via Excitation Power Thresholding
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Solution Overview
Problem
Current luminescence microscopy methods face limitations in achieving high resolution without resorting to multiple wavelengths or complex image reconstruction algorithms, and they struggle with depth resolution when using slit diaphragm systems, which reduces image acquisition speed and introduces crosstalk.
Innovation Solution
A resolution-enhanced luminescence microscopy method that converts a sample from a first luminescent state to a second state with reduced excitability by adjusting the excitation radiation power, using a single light source and a specific excitation radiation distribution with local power maxima and minima, allowing for increased spatial resolution beyond the diffraction limit.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If multiple wavelengths or complex image reconstruction algorithms are used to achieve high resolution, then measurement precision is improved, but device complexity increases
Solution Approach 1:
The patent changes the power parameter of excitation radiation to create a non-linear response in the luminescent sample. By varying excitation power across different spatial zones (below threshold in central zone, above threshold in outer zones), the system achieves super-resolution without requiring multiple wavelengths or complex reconstruction algorithms, thus improving measurement precision while avoiding increased device complexity
Solution Approach 2:
The excitation radiation distribution is segmented into distinct zones with different power levels: a central zone with power below the threshold value and outer zones with power above the threshold value. This segmentation creates spatially differentiated luminescence responses that enable enhanced resolution through simple detection of the boundary between luminescent and non-luminescent regions
2Measurement precision
If slit diaphragm systems are used to improve depth resolution, then measurement precision is improved, but productivity decreases due to reduced image acquisition speed
Solution Approach 1:
The patent extracts the depth resolution function from the mechanical slit diaphragm system and replaces it with an optical threshold-based method. By using the non-linear luminescence response to different excitation powers, the system achieves depth sectioning without physical apertures, thereby maintaining high image acquisition speed while improving depth resolution measurement precision
3Measurement precision
If slit diaphragm systems are used to improve depth resolution, then measurement precision is improved, but device complexity increases due to crosstalk introduction
Solution Approach 1:
The patent converts the potential harm of crosstalk in slit diaphragm systems into a benefit by using threshold-based luminescence switching. The clear binary transition between luminescent and non-luminescent states at the threshold power level creates sharp spatial boundaries that eliminate crosstalk issues, achieving depth resolution with simpler system architecture
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 approach simplifies the microscope design, eliminates the need for multiple wavelengths, and achieves higher resolution and faster image acquisition without depth resolution limitations, enabling sharper images of biological specimens.
Implementation Method 1
a luminescent sample which can be converted from a first luminescence state to a second luminescence state with reduced excitability to emit specific luminescence radiation
Implementation Method 2
the luminescent sample changing from a first luminescence state to a second luminescence state can be converted, in which the sample has reduced excitability to emit the specific luminescence radiation
Data Source
Figure 1a~1b
Figure 2a~2c
Figure 3
AI summary
The method involves displacing two partial regions of a sample in two states, respectively, while irradiation of excitation radiation (A) takes place with an excitation radiation distribution. An image of a luminescent sample has two sample regions in the states, respectively. One of the sample regions prevails for the image of the sample, so that the image has a local resolution which is enhanced in relation to the distribution. An independent claim is also included for a microscope for resolution enhanced luminescence microscopy of a sample.