Single-Cell Dispensing With Image-Guided Well Selection
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Solution Overview
Problem
Existing methods for isolating and analyzing single cells are expensive, labor-intensive, and require large sample inputs, making it difficult to characterize complex diseases like cancer and neurological disorders due to the masking of underlying cell heterogeneity in bulk cell sequencing.
Innovation Solution
A method using Poisson distribution to dispense cells at a single cell per well concentration in a microfluidic device, combined with rapid microscope image analysis to identify and select wells with single cells, and iterative dispensing to achieve desired cell counts in a multi-well device.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If traditional single-cell isolation methods are used, then single cells can be isolated for analysis, but the process becomes expensive and labor-intensive
Solution Approach 1:
The system uses automated robotic liquid handling and image analysis software to perform single-cell identification and selection without manual intervention. The robotic arm automatically dispenses cells based on Poisson distribution, captures images with the microscope, and the software autonomously identifies wells containing single cells, eliminating the need for manual cell counting and selection
Solution Approach 2:
The patent replaces manual mechanical cell isolation methods with an automated optical-mechanical system. A microscope with image capture replaces manual cell inspection, and a robotic liquid handling system replaces manual cell dispensing. The system uses optical fields (microscopy) and automated mechanical arms to achieve what previously required manual labor
2Measurement precision
If traditional single-cell isolation methods are used, then single cells can be isolated, but large sample input is required
Solution Approach 1:
The system extracts only the necessary information (presence/absence of single cells) from the cell suspension without requiring large volumes. By using Poisson distribution to dispense small, controlled volumes into many wells and then imaging to identify which wells contain single cells, the method extracts the needed single cells from a minimal sample input, rather than requiring large-scale sample processing
Solution Approach 2:
The patent performs preliminary dispensing of cells into many wells at once using Poisson distribution, then uses preliminary imaging to identify which wells contain single cells before proceeding to downstream analysis. This preliminary identification step allows the system to work with minimal sample input by pre-sorting cells into individual wells and identifying the correct ones for analysis
3Productivity
If bulk cell population sequencing is performed, then large amounts of data can be obtained, but the underlying heterogeneity of rare cell types is masked
Solution Approach 1:
The patent segments the bulk cell population into individual single cells by dispensing them into separate wells. Instead of analyzing a mixed population together, each cell is isolated in its own well, allowing individual characterization. This segmentation enables the detection of rare cell types and heterogeneity that would be masked in bulk analysis, while still maintaining high productivity through parallel processing of many wells
Data Source
AI summary
The present disclosure provides methods, device, assemblies, and systems for dispensing and visualizing single cells. For example, provided herein are systems and methods for dispensing a dispense volume into a plurality of wells of a multi-well device, where, on average, a pre-determined number of cells (e.g., 1-20) are present in the dispense volume, and determining, via a cellular label, the number of cells present in each of the plurality of wells. Such dispensing and cell detection may be repeated a number of times with respect to wells identified as having less than the pre-determined number of cells in order increase the number wells in the multi-well device containing the desired number (e.g., a single cell).