Circulating Endothelial Cell Detection in Non-Enriched Blood
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Solution Overview
Problem
Current methods for detecting and quantifying circulating endothelial cells (CECs) suffer from variability and lack of sensitivity and specificity, hindering their use as biomarkers for myocardial infarction due to divergent isolation methods and variable immunophenotypical definitions.
Innovation Solution
A method for detecting CECs in non-enriched blood samples by determining the presence of immunofluorescent markers and assessing morphology using fluorescent scanning microscopy, combining distinct immunofluorescent staining and morphological characteristics to identify CECs among nucleated cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If immunomagnetic bead-based capture step is used to enrich rare CEC population prior to identification and quantification, then sensitivity of detection is improved, but device complexity and assay time are increased
Solution Approach 1:
The patent extracts and eliminates the immunomagnetic bead-based enrichment step from the CEC detection workflow. By using automated microscopy with artificial intelligence-based image analysis, the system directly identifies and quantifies CECs in whole blood without requiring preliminary enrichment, thereby simplifying the device while maintaining or improving detection sensitivity.
Solution Approach 2:
The patent replaces the mechanical immunomagnetic capture system with an optical-based automated microscopy system. Instead of using magnetic beads and physical separation, the system uses light microscopy combined with AI image analysis to detect CECs, substituting a complex mechanical enrichment process with a simpler optical detection method.
2Measurement precision
If multiple immunophenotypical markers are used to define CECs, then specificity of identification is improved, but device complexity and assay time are increased
Solution Approach 1:
The patent applies partial action by using a reduced set of immunophenotypical markers compared to traditional methods. Instead of requiring multiple immunostaining steps for different markers, the system uses a minimal panel of markers combined with AI-based morphological analysis to achieve high specificity in CEC identification.
Solution Approach 2:
The patent merges immunophenotypical marker detection with morphological analysis into a unified AI-based detection system. Instead of performing separate immunostaining and morphological assessment steps, the system integrates both approaches into a single automated workflow, reducing complexity while maintaining specificity.
3Ease of operation
If flow cytometry is used to sort and detect CECs, then cell sorting capability is improved, but ability to enumerate very small populations of CECs in presence of abundant WBC population is worsened
Solution Approach 1:
The patent introduces automated microscopy with AI-based image analysis as an intermediary system between blood sampling and CEC detection. This intermediary technology enables direct visualization and enumeration of rare CECs in the context of the complete blood smear, providing both sorting capability and accurate enumeration without the limitations of flow cytometry.
Solution Approach 2:
The patent creates a visual copy of the blood smear through high-resolution microscopy imaging. By capturing detailed images of the blood smear and applying AI-based analysis to these images, the system can identify and enumerate CECs with high accuracy, effectively copying the visual information needed for precise detection without requiring physical cell sorting.
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
The method provides high sensitivity and specificity for classifying subjects as MI patients or healthy controls, enabling early detection of cardiovascular events and facilitating timely treatment.
Implementation Method 1
determining presence or absence of one or more immunofluorescent markers in nucleated cells in the non-enriched blood sample
Data Source
AI summary
The disclosure provides methods for detecting circular endothelial cells (CECs) in a non-enriched blood sample. The present disclosure is based, in part, on the unexpected discovery that CECs can be detected in non-enriched blood samples. The present disclosure is further based, in part, on the discovery that CECs can be detected in non-enriched blood samples by combining the detection of one or more immunofluorescent markers in the nucleated cells of a non-enriched blood sample with an assessment of the morphology of the nucleated cells. The present disclosure is further based, in part, on the discovery that CECs can be detected in non-enriched blood samples by comparing the immunofluorescent marker staining and morphological characteristics of CECs with the immunofluorescent marker staining and morphological characteristics of WBCs. The methods disclosed herein serve to classify human subject in myocardial infarction (MI) patients or healthy controls.