Imaging Flow Cytometry for Cell Nuclei Morphology Sorting
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Solution Overview
Problem
Existing methods for evaluating the quality of isolated cell nuclei are time-consuming and lack a simple, efficient way to assess morphology before committing to downstream molecular biology workflows, often leading to costly sequencing due to unclear nucleus quality.
Innovation Solution
A method using imaging flow cytometry to measure light from isolated cell nuclei, generate images, and assess morphology based on image parameters, with sorting gates determined to separate high-quality nuclei from debris, employing a light detection system and machine learning algorithms.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If high resolution microscopes are used to visualize cell nuclei, then imaging quality is improved, but focusing difficulty and time consumption increase
Solution Approach 1:
The patent replaces manual mechanical focusing with automated image processing algorithms. The system captures multiple images at different focal planes and uses computational methods to generate a single in-focus image, eliminating the need for manual focusing adjustments while maintaining high imaging quality.
Solution Approach 2:
The patent extends the imaging problem from a single 2D focal plane to a 3D volumetric space by capturing images at multiple focal depths. This dimensional extension allows the system to overcome focusing limitations by selecting or combining information from multiple depth layers.
2Productivity
If limited number of images are captured, then acquisition time is reduced, but representativeness of morphology assessment deteriorates
Solution Approach 1:
The patent performs preliminary capture of multiple images at different focal planes before any assessment occurs. This preliminary multi-image acquisition ensures that all necessary morphological information is available, and then uses automated processing to efficiently evaluate the nuclei without requiring manual review of each image.
3Loss of time
If nuclei quality assessment is performed before downstream workflows, then time and cost are reduced, but additional equipment and complexity are required
Solution Approach 1:
The patent integrates multiple functions into a single flow cytometry system: it performs standard particle detection, captures images at multiple focal planes, processes images computationally, and provides morphology assessment - all within one instrument. This multi-functionality avoids the need for separate specialized equipment while achieving comprehensive nuclei quality evaluation.
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 rapid assessment of cell nucleus quality, increasing precision by 5-99% and ensuring 50-99% of sorted nuclei are suitable for downstream assays, reducing time and costs.
Implementation Method 1
measuring light from a sample having isolated cell nuclei in a flow stream
Implementation Method 2
Droplets are passed through an electrostatic field and are deflected based on polarity and magnitude of charge on the droplet
Data Source
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 certain embodiments 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. In some embodiments, sorting gates are 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.