Structured Illumination for Luminescent Imaging
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Solution Overview
Problem
Current sequencing technologies using 'sequencing by synthesis' (SBS) chemistries face limitations in distinguishing between multiple nucleotides due to simultaneous excitation of luminescence sites within a pixel, leading to reduced signal-to-noise ratio and increased costs in miniaturization, as they struggle to efficiently utilize the full potential of imaging pixels.
Innovation Solution
A device and method utilizing a structured illumination source with a multi-beam interference pattern generator, allowing selective excitation of multiple luminescence sites over each imaging pixel at different times, enabling the use of multiple features per pixel and enhancing the ability to distinguish between luminescence signals from different nucleotides.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If multiple fluorescent dyes are used to uniquely identify nucleotides, then the ability to distinguish between nucleotides is improved, but the complexity of the sequencing system increases
Solution Approach 1:
The patent applies periodic action by using time-sequential excitation of different fluorescent dyes. Instead of simultaneously exciting all dyes, the system excites each dye at different time points in a periodic sequence, allowing temporal resolution of multiple nucleotide types while using the same imaging pixel, thus maintaining measurement precision without proportionally increasing device complexity
Solution Approach 2:
The patent segments the excitation process into distinct time intervals, with each interval dedicated to exciting a specific fluorescent dye. This temporal segmentation allows multiple dyes to be used for nucleotide identification while avoiding simultaneous signal overlap, thereby managing system complexity through structured temporal division
2Productivity
If multiple sites are imaged simultaneously within a pixel, then the throughput is improved, but the signal-to-noise ratio deteriorates
Solution Approach 1:
The system uses periodic action to sequentially excite different fluorescent dyes at different time points. This temporal separation ensures that only one dye is excited at a time, eliminating cross-talk and background noise from simultaneously excited dyes, thereby maintaining high signal-to-noise ratio while imaging multiple sites within each pixel over time
Solution Approach 2:
The system performs preliminary action by pre-defining the excitation sequence and timing for each fluorescent dye before data collection. This predetermined temporal structure allows the system to efficiently cycle through multiple dyes and sites, maximizing throughput while ensuring clean separation of signals to maintain signal-to-noise ratio
3Area of stationary object
If imaging pixels are miniaturized to increase density, then the device size is reduced, but the cost of miniaturization increases
Solution Approach 1:
The patent applies universality by enabling each imaging pixel to serve multiple functions: it can detect multiple fluorescent dyes at different time points and image multiple sites within the same pixel over time. This multi-functionality allows the system to maintain high imaging capacity with smaller, less expensive pixels, reducing the cost of miniaturization while preserving device density
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 significantly increases the number of imaged sites per pixel, improving the signal-to-noise ratio and throughput while reducing the cost of miniaturization by allowing for more efficient use of imaging pixels, thereby enhancing the sequencing process.
Implementation Method 1
a first luminophore responsive to the first photons and to emit first luminescence, and a second luminophore responsive to the second photons and to emit second luminescence
Implementation Method 2
A structured illumination source is to direct at least a portion of first photons in an interference pattern having a periodicity corresponding to a pixel spacing in a spatial pattern of a plurality of imaging pixels
Data Source
Figure 1A~1C
Figure 2A~2B
Figure 3A~3B
AI summary
A device includes a plurality of imaging pixels in a spatial pattern with a formation of features disposed over the pixels. A first and a second feature of the formation of features are disposed over a first pixel. A first luminophore is disposed within or over the first feature. A second luminophore is disposed within or over the second feature. A structured illumination source is to direct at least a portion of first photons in an illumination pattern to the first feature at a first time, and to direct at least a portion of second photons in the illumination pattern to the second feature at a second time. The structured illumination source includes an illumination pattern generator having an illumination pattern generator actuator connected to the illumination pattern generator to cause the illumination pattern to translate or rotate relative to the formation of features.