Digital Fluorescence Image Signal-to-Noise Ratio via Autocorrelation
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Solution Overview
Problem
Laser scanning microscopes face a challenge in maintaining a high frame rate while improving the signal-to-noise ratio of digital fluorescence images, as increasing pixel dwell time to enhance the signal-to-noise ratio increases the overall time required to obtain a fluorescence image.
Innovation Solution
The method involves rapid sampling of emission light to generate a sequence of amplitudes per pixel, followed by autocorrelation of these amplitudes with a time offset to determine a correlation amplitude that excludes noise, thereby improving the signal-to-noise ratio without reducing the frame rate.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If pixel dwell time is increased to improve signal-to-noise ratio, then measurement precision is improved, but frame rate decreases
Solution Approach 1:
The patent segments the amplitude measurement process into multiple individual amplitude measurements taken at different time points during the pixel dwell time, rather than taking a single average measurement. This segmentation allows the system to capture temporal variations in the fluorescence signal and apply autocorrelation analysis to distinguish signal from noise, thereby improving signal-to-noise ratio without requiring increased dwell time and maintaining high frame rates
Solution Approach 2:
The patent implements autocorrelation analysis of the amplitude sequence, where each amplitude measurement is correlated with subsequent measurements. This feedback mechanism uses the temporal structure of the signal to identify and enhance true fluorescence signal while suppressing random noise, achieving improved measurement precision without extending the pixel dwell time and thus preserving high frame rates
2Measurement precision
If pixel dwell time is increased to improve signal-to-noise ratio, then measurement precision is improved, but time consumption increases
Solution Approach 1:
The patent divides the pixel dwell time into multiple discrete measurement points, taking several rapid amplitude measurements rather than a single prolonged measurement. This segmentation enables statistical analysis through autocorrelation to improve signal-to-noise ratio while keeping the total measurement time short, thus reducing time loss in image acquisition
Solution Approach 2:
The patent performs autocorrelation analysis on the sequence of amplitude measurements to pre-identify the characteristic signal pattern before final image reconstruction. This preliminary action of noise filtering through autocorrelation allows the system to achieve high measurement precision without extending pixel dwell time, thereby minimizing time consumption in obtaining the fluorescence image
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 results in a digital fluorescence image with a higher signal-to-noise ratio, allowing for improved image quality without increasing the total time required to capture the image, as the noise is virtually excluded from the total amplitude calculation.
Implementation Method 1
Fluorescence microscopy is a highly effective light microscopy technique for localizing proteins and imaging protein distributions within tissue samples and cells
Implementation Method 2
Illumination with light (excitation light) of a specific wavelength (excitation wavelength), depending on the dye used, excites the proteins, raising electrons in the dye molecules (fluorophores) to a higher energy level. After a short dwell time, the excited electrons return to their original energy level by emitting light (emission light) of a specific wavelength (emission wavelength) that is longer than the excitation wavelength
Data Source
Figure 1~3b
AI summary
In a method for creating a digital fluorescent image, the light emitted per pixel from an object plane is converted into a sequence of amplitudes (I), each of which is associated with one specific measurement time, the sequence of amplitudes (I) is auto-correlated in a manner that is delayed by at least one time offset (τ), and a specific correlation amplitude (Kτ), from which a total amplitude is determined, is formed for each of the time offsets (τ).