3D Imaging Flow Cytometry With Light-Sheet Scanning
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Solution Overview
Problem
Conventional imaging flow cytometry systems are limited to 2D cell imaging, leading to occlusion of objects, blurring due to focal depth, loss of z-axis spatial resolution, and artifacts from projecting 3D cells into 2D images, which hinders accurate phenotyping and biological insights.
Innovation Solution
A three-dimensional imaging flow cytometry (3D-IFC) system using a light sheet fluorescence microscopy configuration with an acousto-optic deflector for high-speed scanning in the z-direction, combined with a spatial filter and single-element photodetector to detect fluorescence optical signals, allowing spatial-to-temporal transformation for rapid 3D image reconstruction.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional 2D imaging flow cytometry is used, then the system structure is simple and operation is easy, but z-axis spatial resolution is lost and occlusion occurs
Solution Approach 1:
The patent transitions from 2D imaging to 3D imaging by introducing a light sheet illumination system that illuminates the sample in the z-direction and uses a scanning mechanism to acquire optical signals from multiple focal depths. This dimensional expansion enables z-axis spatial resolution while maintaining manageable system complexity through modular design
Solution Approach 2:
The 3D imaging process is segmented into multiple focal depth planes that are sequentially scanned along the z-axis. Each plane is imaged separately and then reconstructed into a complete 3D volume, allowing precise z-axis resolution without requiring the entire 3D volume to be captured simultaneously, thus managing device complexity
2Measurement precision
If conventional 2D imaging is used, then the device complexity is low, but occlusion of objects and blurring due to focal depth occur
Solution Approach 1:
By adding the z-dimension through light sheet illumination and focal depth scanning, the system captures the complete spatial distribution of particles without occlusion. Objects at different depths are resolved separately and reconstructed in 3D space, eliminating the blurring and occlusion inherent in 2D projection
Solution Approach 2:
The light sheet acts as an intermediary that selectively illuminates specific focal planes while the scanning mechanism mediates the acquisition of signals from different depths. This intermediary approach enables precise spatial mapping without direct complex interaction between all system components
3Measurement precision
If high-speed scanning is implemented for 3D imaging, then 3D spatial resolution is improved, but the complexity of the illumination system increases
Solution Approach 1:
The illumination system uses periodic scanning of the light sheet along the z-axis at high speed. This periodic action allows the system to acquire multiple focal planes in rapid succession, achieving 3D spatial resolution through time-multiplexed illumination rather than requiring complex simultaneous multi-plane illumination
Solution Approach 2:
The patent replaces complex mechanical scanning systems with an acousto-optic deflector (AOD) that uses acoustic fields to control light deflection. This substitution reduces mechanical complexity while enabling high-speed scanning and precise 3D spatial resolution through non-mechanical beam steering
4Measurement precision
If 3D image reconstruction is performed, then accurate phenotyping is achieved, but data processing time and complexity increase
Solution Approach 1:
The system performs preliminary organization of optical signals during the scanning process itself, mapping signals to their corresponding spatial coordinates and focal depths as they are acquired. This preliminary structuring of data reduces the computational burden during final 3D reconstruction, enabling accurate phenotyping with reduced processing time
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
Enables high-throughput, continuous 3D image acquisition of cells in flow, providing accurate 3D spatial distribution of fluorescence and scattering light for improved phenotyping and biological insights, such as DNA damage analysis and FISH signal detection.
Implementation Method 1
a light source to produce a light beam that is optically coupled to light redirection device to modify the light beam by redirecting the light beam to different angles
Implementation Method 2
a spatial filter positioned between the particle motion device and the one or more photodetectors, the spatial filter including a plurality of apertures to selectively allow a portion of the asymmetric illumination area of light
Implementation Method 3
an optical detection system optically interfaced with the particle motion device and operable to obtain optical signal data associated with different parts of the particle
Data Source
Figure 1A~1B
Figure 1C~1D
Figure 2A~2D
AI summary
Disclosed are methods, devices, systems and applications for camera-less, high-throughput three-dimensional imaging of particles in motion. In some aspects, a system includes a particle motion device to allow particles to move along a travel path; an optical illumination system to produce an asymmetric illumination area of light in a region of the travel path of a particle that scans over a plurality of sections of the particle at multiple time points while the particle is moving; an optical detection system optically interfaced with the particle motion device to obtain optical signal data associated with different parts of the particle corresponding to the particle's volume during motion in the travel path; and a data processing unit to process the optical signal data obtained by the optical detection system and produce data including information indicative of 3D features of the particle.