Microelectrophoresis platform for fast serial analysis of single cells
Dechen Jiang1, Christopher E Sims, Nancy L Allbritton
1Department of Chemistry, University of North Carolina, Chapel Hill, NC, USA.
Electrophoresis
|July 7, 2010
Summary
This study introduces a capillary electrophoresis system for rapid single-cell analysis. The platform enables fast serial analysis of individual cells, improving throughput for cellular studies.
Area of Science:
- Analytical Chemistry
- Biotechnology
- Cell Biology
Background:
- Single-cell analysis is crucial for understanding cellular heterogeneity.
- Existing methods for serial single-cell analysis can be time-consuming.
- Microfluidic platforms offer potential for high-throughput cellular assays.
Purpose of the Study:
- To develop a capillary-based microelectrophoresis platform for fast serial analysis of single cells.
- To enable rapid, sequential analysis of individual cells with defined locations.
- To characterize the performance and throughput of the developed system.
Main Methods:
- A capillary electrophoresis system with a two-channel flow system for buffer switching.
- Utilizing an array of cell microwells for retaining and sequentially delivering single cells.
- Employing a focused laser pulse for cell lysis and subsequent electrophoretic separation.
Main Results:
- Demonstrated serial analysis of 28 single cells within 16 minutes, achieving a rate of 1.8 cells/min.
- Characterized key system parameters including fluid flow rates, cell array dimensions, and laser energies.
- Identified peak overlap between successive samples as a limitation to current throughput.
Conclusions:
- The developed capillary-based microelectrophoresis platform facilitates fast serial analysis of single cells.
- The system's design allows for sequential cell delivery and lysis for electrophoretic separation.
- Further optimization is needed to overcome throughput limitations caused by sample peak overlap.
Related Concept Videos
01:20Electrophoresis: Overview
Electrophoresis is a powerful analytical separation technique that relies on the differential migration of charged species when subjected to an electric field. The core strength of electrophoresis lies in its ability to separate high-molecular-weight species in complex mixtures. It has found widespread use in biochemistry, molecular biology, and analytical chemistry, allowing the separation of compounds like amino acids, nucleotides, carbohydrates, and proteins with excellent resolution.
There...
There...
01:30Capillary Electrophoresis: Applications
Capillary electrophoretic separations offer various modes, each with unique applications. These modes include capillary zone electrophoresis, capillary gel electrophoresis, capillary array electrophoresis, capillary isoelectric focusing, capillary isotachophoresis, micellar electrokinetic chromatography, and capillary electrochromatography.
Capillary zone electrophoresis (CZE) separates ionic components based on their electrophoretic mobility. It has been used to separate proteins, amino acids,...
Capillary zone electrophoresis (CZE) separates ionic components based on their electrophoretic mobility. It has been used to separate proteins, amino acids,...
01:20Capillary Electrophoresis: Instrumentation
Capillary electrophoresis instrumentation typically consists of several key components. A high-voltage power supply generates the electric field necessary for the separation by connecting to an anode (the positively charged electrode) and a cathode (the negatively charged electrode) located in buffer reservoirs at each end of the capillary tube. The system includes a sample vial, a fused silica capillary tube coated with polyimide for mechanical strength through which the sample components...
01:22Two-dimensional Gel Electrophoresis
Two-dimensional gel electrophoresis is a high-resolution protein separation method first introduced by O' Farrell and Klose in 1975. This method involves protein separation by two dimensions, mass and charge, making it more accurate than one-dimensional gel electrophoresis.
The first dimension separation uses the isoelectric focusing or IEF technique performed on immobilized pH gradient (IPG) strips that separate proteins according to their isoelectric points.
Biological samples, such as cells...
The first dimension separation uses the isoelectric focusing or IEF technique performed on immobilized pH gradient (IPG) strips that separate proteins according to their isoelectric points.
Biological samples, such as cells...