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Microscale Vortex-assisted Electroporator for Sequential Molecular Delivery
Published on: August 7, 2014
Continuous and quantitative delivery of molecules into individual cells with a diffusional microburet
Miklós Gratzl1, Chen Yi, Gary R Bright
1Department of Biomedical Engineering, Case Western Reserve University, Cleveland, Ohio 44106, USA. mxg13@case.edu
Analytical Chemistry
|June 25, 2009
Summary
This study introduces the diffusional microburet (DMB) for sustained, direct delivery of molecules into live cells. This method enables precise intracellular dosing for various biological and clinical research applications.
Area of Science:
- Cell Biology
- Biotechnology
- Pharmacology
Background:
- Direct delivery of molecules into the cytosol of live cells is crucial for biological and clinical research.
- Current methods may lack precision or require significant volume changes within the cell.
Purpose of the Study:
- To develop a novel method for continuous and controlled molecular delivery into single live cells.
- To enable quantitative analysis of intracellular processes through sustained dosing.
Main Methods:
- Development of a diffusional microburet (DMB), a micropipette with a hydrogel plug.
- Utilizing a concentration gradient for diffusive delivery of molecules into the cytosol.
- Demonstration of dosing Lucifer Yellow CH into a7r5 vascular smooth muscle cells.
Main Results:
- The DMB provides sustained, reproducible, and quantifiable intracellular dosing without altering intracellular volume.
- Hydrogel properties can minimize biomolecule loss from the cytosol.
- Intracellular concentrations of fluorescent molecules can be determined without calibration.
Conclusions:
- The DMB technology offers a new platform for precise molecular delivery and quantitative analysis in single live cells.
- This method facilitates experiments such as intracellular ion titration and drug resistance pathway analysis.
