Microfluidic Micropillar Capture for Sequential Multiomic Isolation
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Solution Overview
Problem
Current methods for multiomic analysis of cell analytes are inefficient, insensitive, and lack comprehensive capabilities for studying total protein, plasma membrane protein, nuclear RNA and protein, cytosolic RNA and protein, chromatin, and genomic DNA, particularly for detecting genetic mutations in cancer and genetic disorders.
Innovation Solution
A microfluidic device with cell capture and nucleic acid entanglement components, utilizing micropillars, performs continuous flow to isolate distinct analyte components like total protein, plasma membrane protein, total RNA, cytosolic RNA, nuclear RNA, nuclear protein, chromatin, gDNA markers, amplified gDNA, and methylated gDNA through sequential workflow protocols under controlled conditions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If traditional methods are used for multiomic analysis, then the analysis can be performed, but the efficiency is low and sensitivity is insufficient
Solution Approach 1:
The patent divides the cell into distinct compartments (nuclear and cytosolic) and separates different analyte types (proteins, RNA, DNA) into different fractions. The microfluidic device segments the analysis process into sequential workflow steps, each isolating specific analytes from the cell sample, enabling efficient and sensitive multiomic analysis.
Solution Approach 2:
The microfluidic device performs multiple functions within a single integrated system: cell capture, nucleic acid entanglement, sequential analyte isolation, and fraction collection. This multi-functional platform enables comprehensive multiomic analysis of various analytes (total protein, plasma membrane protein, nuclear RNA, cytosolic RNA, chromatin, gDNA) simultaneously, greatly improving efficiency and sensitivity.
2Adaptability or versatility
If comprehensive multiomic analysis is performed, then all analyte components can be studied, but the device complexity increases
Solution Approach 1:
The patent employs nested micropillar structures where nucleic acid entanglement micropillars are positioned within or alongside cell capture micropillars. This nested arrangement allows the device to handle multiple analyte types (DNA, RNA, proteins) in a compact, hierarchical structure, achieving comprehensive coverage without proportionally increasing device complexity.
Solution Approach 2:
The device performs preliminary cell capture and nucleic acid entanglement before the sequential analyte isolation workflow. By pre-positioning cells and nucleic acids in the microfluidic device, the system prepares all necessary components in advance, enabling comprehensive multiomic analysis through a standardized sequential protocol that reduces overall system complexity.
3Measurement precision
If sequential workflow procedure is used, then distinct analyte components can be isolated, but the analysis time increases
Solution Approach 1:
The microfluidic device operates under continuous flow conditions throughout the sequential workflow procedure. This continuous flow enables simultaneous progression of multiple isolation steps, maintains consistent hydrodynamic conditions for reliable analyte separation, and eliminates idle time between steps, thereby reducing total analysis time while maintaining high separation purity.
Solution Approach 2:
The patent utilizes hydrodynamic flow control in the microfluidic device to drive the sequential isolation of analytes. By precisely controlling flow rates and hydraulic conditions, the system achieves efficient analyte separation and rapid fraction collection, minimizing analysis time while maintaining the purity and integrity of distinct analyte components.
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 efficient and comprehensive isolation of multiple analyte components suitable for further analysis, enhancing the understanding of genetic mutations in cancer and genetic disorders.
Implementation Method 1
introducing the one or more cell sample into the microfluidic device under sufficient hydrodynamic flow to entrap, by size exclusion, the one or more cell within the cell capture component
Implementation Method 2
entrap, by size exclusion, the one or more cell within the cell capture component
Implementation Method 3
treating the one or more captured cell with a sequential workflow procedure under conditions effective to separate one or more distinct analyte component therefrom
Data Source
AI summary
A method for isolating analytes is disclosed. The method involves: introducing a sample including a cell into a microfluidic device, where the microfluidic device includes cell capture micropillars; entrapping the cell among the cell capture micropillars; separating at least two analytes selected from the group consisting of (i) protein, (ii) RNA, and (iii) DNA; and isolating the separated analytes.