Trajectory-Based Triggering for Hyperspectral Flow Cytometry
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Solution Overview
Problem
Current high throughput instrumentation in cytometry lacks the capability to quickly screen and sort large complex cell populations using hyperspectral imaging, which is essential for advanced biological investigations and drug development.
Innovation Solution
A hyperspectral imaging flow cytometer is developed, incorporating a microfluidic flow system, a trajectory-based triggering system, and a hyperspectral confocal imaging system that allows for the acquisition of high-resolution images of particles flowing through a channel, enabling real-time multivariate analysis and sorting of cells based on spectral and spatial signatures.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If hyperspectral confocal imaging system is used to acquire high-resolution images of flowing particles, then measurement precision is improved, but productivity deteriorates due to time-consuming sequential scanning
Solution Approach 1:
The trajectory-based triggering system performs preliminary tracking of particle positions using wide-field imaging, predicts future particle locations, and pre-positions the confocal laser scan. This preliminary action allows the high-resolution imaging to be performed exactly when particles are in the optimal position, eliminating idle scanning time and achieving both high resolution and high throughput simultaneously
2Device complexity
If traditional wide-field triggering system is used, then device complexity is reduced, but measurement precision deteriorates due to inability to accurately track and image individual particles at optimal positions
Solution Approach 1:
The triggering system is segmented into two functional components: a wide-field imaging subsystem for particle detection and trajectory prediction, and a confocal imaging subsystem for high-resolution spectral acquisition. This segmentation allows each subsystem to be optimized for its specific function while working together to achieve both simplicity and precision
3Productivity
If high flow velocity is used to increase throughput, then productivity is improved, but measurement precision deteriorates due to reduced time for image acquisition and analysis
Solution Approach 1:
The system replaces mechanical scanning with a trajectory-based triggering mechanism that uses computational prediction of particle positions. By substituting the mechanical scanning approach with a smart triggering system that anticipates particle locations, the system can maintain high flow velocities while ensuring high-resolution images are captured at the optimal moment, preserving both throughput and precision
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 system enables the rapid characterization of thousands of cells per session, providing detailed spatial maps and spectral information, thereby enhancing throughput and accuracy in biological research and drug development.
Implementation Method 1
obtaining a first image of a particle a first time at a first location in the channel, obtaining a second image of the particle at a later second time at a second location in the channel, predicting a third time and a lateral (i.e., transverse or perpendicular to the fluid flow direction) location at which the particle will cross an imaging line
Implementation Method 2
acquiring a hyperspectral image of fluorescence emitted by the particle in the imaging line
Implementation Method 3
laterally scanning a focused laser beam along the imaging line
Implementation Method 4
hyperspectral confocal imaging system having a focal plane downstream from the particle detection system
Implementation Method 5
microfluidic flow system for injecting a sample of fluorescent particles into a channel, directing the particles to flow through an imaging field in the channel
Data Source
AI summary
A hyperspectral imaging flow cytometer can acquire high-resolution hyperspectral images of particles, such as biological cells, flowing through a microfluidic system. A trajectory-based triggering system can be used that will only trigger the acquisition of a hyperspectral image when an appropriate particle or cell is crossing an imaging line, thereby saving valuable resources and time. The hyperspectral imaging flow cytometer can provide detailed spatial maps of multiple emitting species, cell morphology information, and state of health. An optimized system can image about 20 cells per second. The hyperspectral imaging flow cytometer enables many thousands of cells to be characterized in a single session.