Traceable Impedance-Based Dispensing and Cloning of Living Single Cells

Impedance-Based Single-Cell Pipetting

“…We characterized this assumption elsewhere using a wide range of cell types. 35 Finally, a cell viability assay using fibroblast cells indicated that the instrumented pipet has no detrimental effect on cells when compared with standard pipetting. As described by Zimmermann et al 36 in an electric-based cell detection system, the electric field applied to cells can change cell viability by breaking down the lipid membrane.…”

“…Higher electric fields would induce a permanent destruction of the cell membrane leading to cell death. 35,36 Both effects need to be avoided in the instrumented pipet. First, even though the reversible opening should not lead to cell apoptosis, 36 a bias would be induced in the effective impedance change measurement because current would flow through the cell 35 and thus induce an error on the cell size interpretation.…”

“…35,36 Both effects need to be avoided in the instrumented pipet. First, even though the reversible opening should not lead to cell apoptosis, 36 a bias would be induced in the effective impedance change measurement because current would flow through the cell 35 and thus induce an error on the cell size interpretation. Second, this effect could also lead to the uptake of external molecules by the cell as it occurs during electroporation, 36 which is to be avoided here.…”

A new workflow combining magnetic cell separation and impedance-based cell dispensing for gentle, simple and reliable cloning of specific CD8+ T cells

“…Consequently, re-cloning steps are warranted which comes as money and time consuming. Therefore, we developed a novel 2-step workflow combining simple, affordable and gentle magnetic cell separation followed by single cell isolation using a device called DispenCell-S1 9,10 . Moreover, this device provides an immediate proof of single cell isolation, which guarantees the monoclonality of selected T cell candidates and reduce the need for checking cell clonality.…”

Traceable impedance-based single-cell pipetting, from a research set-up to a robust and fast automated robot: DispenCell-S1

“…High producing clonallyderived cell lines are however rare events and their identification is a time-consuming process that requires the screening of thousands of clones. Therefore, there is an unmet need for a device that would allow the fast and efficient isolation of single cells, while preserving their integrity and providing an insurance of their clonality.We proposed earlier an impedance based pipetting technology for isolation of single cells (Bonzon et al, 2020), with initial validations for state-of-the-art stem cell in-vitro and in-vivo assays (Muller et al, 2020). Here, we present the transition from this pioneering technology developed in an academic setting into an automated instrument, called DispenCell-S1, allowing for traceable isolation of single cells.We developed and validated models predicting the performances for 96-well plates single-cell isolation.…”