Cell Nuclei Flow Imaging for Rapid Quality Sorting
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Solution Overview
Problem
Current methods for evaluating the quality of isolated cell nuclei are time-consuming and lack simplicity, often leading to costly downstream molecular biology workflows without adequate assurance of nuclei quality.
Innovation Solution
A method and system for assessing cell nuclei morphology using light measurement and image analysis, enabling rapid determination of nuclei viability, ploidy, size, shape, and elasticity, with sorting gates based on image parameters and machine learning algorithms to separate high-quality nuclei from debris.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If high resolution microscopes (60X) are used to visualize cell nuclei, then visualization quality is improved, but focusing becomes challenging and the limited number of images may not be representative of the morphology and quality of isolated nuclei
Solution Approach 1:
The patent replaces the mechanical focusing system of traditional microscopes with an optical diffraction-based imaging system. Instead of using high-resolution microscopes requiring manual or automated focusing mechanisms, the invention uses a flow cytometer with a 488 nm laser to illuminate nuclei in a fluid stream, capturing images based on light diffraction patterns. This substitution eliminates the complex focusing mechanism while maintaining imaging capability.
Solution Approach 2:
The patent utilizes fluid dynamics to transport nuclei through the imaging system. Nuclei are suspended in a fluid stream that flows through a detection region where images are captured. The hydraulic flow system automatically positions nuclei in the imaging plane, replacing the need for mechanical focusing adjustments and enabling continuous imaging of numerous nuclei for statistical representativeness.
2Measurement precision
If traditional quality assessment methods are used before downstream molecular biology workflows, then thorough evaluation is achieved, but the process becomes time-consuming and costly
Solution Approach 1:
The patent implements continuous imaging and analysis of nuclei as they flow through the detection region. The flow cytometer captures images of nuclei continuously at high speed, allowing rapid assessment of morphology parameters including size, shape, and chromatin organization. This continuous action enables thorough quality evaluation without the time delays inherent in traditional batch processing methods.
Solution Approach 2:
The patent creates a controlled fluid environment that preserves nuclei integrity during rapid analysis. The buffered fluid stream maintains physiological conditions, preventing degradation or morphological changes during the quick imaging process. This inert environment allows fast assessment without compromising the accuracy of quality evaluation.
3Device complexity
If limited number of images are taken with high resolution microscope, then device complexity is reduced, but the images may not be representative of the morphology and quality of isolated nuclei
Solution Approach 1:
The patent uses continuous flow imaging to capture hundreds or thousands of nuclei images rapidly. The fluid stream continuously delivers nuclei through the detection region, enabling high-throughput imaging without complex device changes. This large sample size ensures statistical representativeness of the morphology and quality assessment.
Solution Approach 2:
The patent creates multiple optical copies of nuclei images as they pass through the detection region. Each nucleus is imaged multiple times during its passage, and numerous nuclei are imaged in sequence. These multiple copies provide redundant data that enhances the reliability and representativeness of the quality assessment while using a relatively simple imaging system.
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
Enhances the precision of determining nuclei quality by 5-99%, allowing fast sorting of high-quality nuclei for downstream assays, increasing the suitability of sorted nuclei for downstream applications by 50-99%.
Implementation Method 1
measuring light from a sample having isolated cell nuclei in a flow stream
Implementation Method 2
measuring light from a sample having isolated cell nuclei in a flow stream
Implementation Method 3
Droplets are passed through an electrostatic field and are deflected based on polarity and magnitude of charge on the droplet into one or more collection containers
Data Source
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AI summary
Aspects of the present disclosure include methods for assessing morphology of isolated cell nuclei (e.g., to determine viability of the cell nuclei) in a sample. Methods according to the present disclosure include measuring light from a sample having isolated cell nuclei in a flow stream, generating an image of the cell nuclei from the measured light and assessing morphology of the cell nuclei based on the generated images of the cell nuclei. Sorting gates can be determined based on images or image parameters calculated for the cell nuclei in the sample. Systems and integrated circuit devices (e.g., a field programmable gate array) for practicing the subject methods are also described. Non-transitory computer readable storage medium are also provided.