Flow Cytometer Optical Layout for Multi-Dye Spectral Detection
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Solution Overview
Problem
Conventional flow cytometers face limitations in simultaneously detecting multiple fluorescent dyes with similar emission spectra due to complex optical designs, high cost, and reduced light collection efficiency, especially at high sample flow rates, leading to large signal coefficient of variation and increased maintenance needs.
Innovation Solution
An optical system with shared light collection and magnification optics, combined with a single set of light splitting filters, uses spherical lenses to direct and split light into discrete wavelength ranges, eliminating the need for optical fibers and reducing mechanical interference, thereby improving light collection efficiency and stability.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If optical fibers are used to collect fluorescent light from each laser, then light collection is achieved, but the system becomes complex and costly with N optical fibers needed for N lasers
Solution Approach 1:
The patent combines multiple optical fibers into a single bundled fiber optic cable that collects light from multiple lasers simultaneously. This merging approach maintains the light collection capability while significantly reducing system complexity and the number of individual optical components needed.
Solution Approach 2:
The bundled optical fiber system serves multiple functions by collecting fluorescent light from all lasers through a single unified structure. This universal approach eliminates the need for separate optical collection paths for each laser, reducing overall system complexity.
2Measurement precision
If multiple optical components are used for dispersing or splitting light, then spectral analysis is achieved, but the system becomes very complex and costly
Solution Approach 1:
The patent extracts the light dispersion function from complex multi-component optical systems and implements it through a simplified prism-based approach. This extraction maintains spectral analysis capability while dramatically reducing the number of required optical components.
Solution Approach 2:
The patent replaces expensive, complex optical dispersion components with a more economical prism-based system that achieves the same spectral separation function with fewer and less costly components.
3Ease of operation
If conventional flow cytometers use discrete channels for each fluorescent dye, then detection is simplified, but fluorescent dyes with similar emission spectra cannot be used simultaneously
Solution Approach 1:
The patent segments the continuous spectral range into multiple detection channels using optical filters, allowing simultaneous detection of multiple fluorescent dyes with similar emission spectra. This segmentation enables the system to distinguish between dyes that would be indistinguishable in conventional single-channel systems.
Solution Approach 2:
The patent transitions from conventional single-channel detection to multi-dimensional spectral analysis by collecting light across a wide wavelength range and using de-convolution algorithms to resolve overlapping spectra. This dimensional expansion enables simultaneous detection of multiple fluorescent dyes.
4Ease of manufacture
If light collection optics are designed with low magnification, then coupling into optical fibers is easier, but light collection efficiency is reduced at high flow rates
Solution Approach 1:
The patent optimizes the magnification parameter of the light collection optics to balance two competing requirements: sufficient magnification to maintain light collection efficiency at high flow rates, and appropriate coupling to the optical fiber bundle. This parameter optimization achieves both ease of manufacture and high productivity.
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 high sensitivity and reduced cost by maximizing light collection, minimizing signal variation, and simplifying the design, while maintaining consistent fluorescent signal detection across multiple lasers, even at high flow rates.
Implementation Method 1
each of the spherical lenses within the array configured to receive the magnified light emitted from one of the different positions of the flow cell and to direct the received magnified light to a predetermined area on a corresponding photodetector
Implementation Method 2
shared light collection optics and shared magnification optics configured to collect light emitted from the different positions of the flow cell and to magnify and focus the collected light on a focal plane
Implementation Method 3
an array of photodetectors configured to measure the magnified light directed from each of the spherical lenses
Implementation Method 4
As cells pass through the flow cell, they are struck by a light source tuned to a frequency that causes the fluorescent dye to fluoresce
Data Source
AI summary
Systems and methods for measuring light, which collect and measure light emitted from different positions along the length of a flow cell using shared collection and magnification optics, a light splitting module that splits magnified light into a plurality of wavelength ranges, and an array of spherical lenses that direct light of different wavelength and emission position to an array of detectors.