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A Rapid, Scalable Method for the Isolation, Functional Study, and Analysis of Cell-derived Extracellular Matrix
Published on: January 4, 2017
The VerIFAST: an integrated method for cell isolation and extracellular/intracellular staining
Benjamin P Casavant1, David J Guckenberger, Scott M Berry
1Department of Biomedical Engineering, Wisconsin Institutes for Medical Research, University of Wisconsin-Madison, Madison, WI, USA.
Lab on a Chip
|December 11, 2012
Summary
The VerIFAST device integrates cell isolation with staining, reducing sample loss for rare cell studies. This method achieves high capture efficiency and purity without transfers or centrifugation.
Area of Science:
- Biotechnology
- Cell Biology
- Analytical Chemistry
Background:
- Studying rare cell subpopulations requires efficient isolation methods.
- Traditional methods often involve transfer and wash steps, leading to sample loss.
- Integrated approaches minimize loss, crucial for rare cell analysis.
Purpose of the Study:
- To introduce and characterize the VerIFAST device for integrated cell isolation and staining.
- To demonstrate the device's efficiency and purity in capturing rare cells.
- To showcase the capability for multi-step cellular staining within the device.
Main Methods:
- Utilized the Immiscible Filtration Assisted by Surface Tension (IFAST) principle for simplified workflow.
- Incorporated a front-end purification step for passive settling and capture of cells.
- Employed a Sieve Chamber for removal of paramagnetic particles (PMPs) and multi-step washing without centrifugation.
- Performed extracellular and intracellular staining for EpCAM, pan-cytokeratins, and Ki-67.
Main Results:
- Achieved high capture efficiency (>80%) and purity (>70%) in the isolation step.
- Demonstrated successful removal of unbound PMPs and complex washing procedures.
- Successfully performed extracellular and intracellular staining within the VerIFAST device.
Conclusions:
- The VerIFAST device offers an integrated solution for rare cell isolation and characterization.
- The device minimizes sample loss through its transfer- and centrifugation-free workflow.
- VerIFAST enables efficient cell isolation coupled with complex staining protocols for downstream analysis.
