Platform for Advanced Single Cell-Manipulation and Analysise
Abstract: Living biological cells and cell cultures are the essential ingredient of a wide array of life sciences research, from the genetic basis and mechanism of disease to drug development and drug‐cell interactions. So far, cells cannot be easily isolated and systematically analysed individually. They are commonly manipulated only as an unordered, unsorted, heterogeneous cell assembly often after lengthy periods of cell culture and isolation. This limits the depth of perspective that can be gained using modern single cell analysis methods, on single cell interactions and on the behaviour of a single cell types in a mixed cell or tissue system. The Single Cell Manipulation (SCM) technology investigated within the PASCA project, promises to open up new opportunities for enhanced life science research, especially in areas such as cell biology, stem cell research, cancer research, drug development and for the fabrication of 3D cell cultures or artificial organs.
The overall aim of the PASCA project is to establish and validate a platform for single cell experiments. The project has two main objectives; firstly to develop a cell printing instrument for the manipulation of single living cells so that individual cells suspended in a heterogeneous cell solution could be isolated, transported and individually relocated and rearranged onto an arbitrary target. This approach requires the development of technology for printing single cells confined in micro droplets of only hundred micron size in an “inkjetlike” manner, so that the number of cells per droplet can be monitored and controlled by optical or impedance sensors. The SCM prototype instrument developed on the basis of this approach. enables the identification, sorting and delivery of single cells in an “ink‐jet like” manner, contained in a picoliter sized droplet for downstream processes such as transfection, stimulation, culturing or analysis.
The second major objective of the project is to apply the single cell manipulation (SCM) technology to a set of applications thereby establishing a platform for advanced single cell manipulation and analysis (PASCA). Within this platform approach, the SCM technology is complements the "upstream" processes of cell preparation and treatment and the "downstream" processes of transfection, cell incubation and analysis, to realize and demonstrate a complete process flow for specific single cell experiments. Though the process flow for a certain experiment is specific, the approach of combining the SCM with upstream and downstream technologies in a flexible way is generic and can be adapted to many other applications.. Thus, by using the SCM technology coupled to state‐of‐the‐art upstream and downstream technologies (e.g. patch clamping, next generation sequencing, etc.), a novel platform for single cell based experiments ‐ referred to as the PASCA platform ‐ was established and validated.