High-throughput Cell-capture Device for Semi-automated Single-cell Nanobiopsy and Nanomanipulation
Engineering
BME 195
A single-cell nanomanipulation system allows the user to investigate the cellular processes of a single cell, over a period of time, without killing the cell. This has been done on cells that are stuck to the bottom of a petri dish. To investigate cells that are usually floating, requires a means of capturing the cells out of solution and holding them down in a way that allows a nanopipette to access the cells.
The focus of this research is a microfluidic cell-capture device. I characterized and integrated a cell-capture device with the single-cell nanomanipulation system. The device consists of a sandwich structure with a transparent membrane window perforated by a 6 X 6 array of 2-micrometer (µm) diameter through-holes. The device, coupled with a negative-pressure control system, enables the capture and release of single cells floating in cell media while keeping the cells alive. By supplying a negative pressure of 200 Pascals (Pa), the device captures cells in cell media, immobilizes the cells at the through-holes, and releases the cells on demand.
As a proof of concept, HeLa cells expressing green fluorescent protein were captured on the device and nanobiopsies were attempted on 12 single cells to aspirate subpopulations of mitochondria. I conducted polymerase chain-reaction to amplify mitochondrial DNA (mtDNA) from each sample. Initial results are inconclusive, but remain promising. Ongoing research will further develop and characterize the cell-capture device.
