Single Cell Analysis and FluidFM Technology
The ability to obtain information at the single-cell level and to perturb individual cells is of central importance for numerous biological questions. There is a growing awareness that individual cells, although genetically identical to sister cells, show different phenotypes and gene expression profiles and therefore different levels of proteins and metabolites. There is also a need to manipulate individual cells to validate hypotheses and to substantiate fundamental biological principles. The analysis of individual cells represents a major challenge, and thus, there is a demand for innovative technologies for cell perturbation as well as sampling, extraction, and high-sensitivity molecular detection.
The recently invented fluidic force microscope (FluidFM) by T. Zambelli and coworkers (ETH Zurich, D-ITET) is an atomic force microscope (AFM) provided with microchanneled cantilevers for local liquid dispensing (Meister et al., 2009 Nano Letters 9:2501–7). The "nanosyringe" is operated on top of an inverted optical microscope, allowing optical inspection during cell manipulation. In an interdisciplinary collaboration with the inventors of the FluidFM and the ETH Spinoff company Cytosurge, we are developing the technology for applications in different complementary areas of single-cell manipulation, including the spatial manipulation, cell adhesion, injection, and extraction of single cells. We established spatial manipulation of viruses, bacteria, yeasts and mammalian cells and are also using the technology to study host-microbe interactions. Single-cell force spectroscopy (SCFS) of individual cells has been demonstrated for bacteria, yeasts and mammalian cells. During this process, the chemical fixation used in conventional SCFS is replaced by sucking the cell to the cantilever opening by underpressure, allowing the generation of independent biological data in a serial fashion within a relatively short period of time. In another line of experiments, we demonstrated the controlled injection of femtoliter volumes into single mammalian cells and dedicated subcompartments, i.e., the nucleus. Among other potential applications, experimental perturbation by nanoinjection with the FluidFM can target the metabolism and other subsystems of the cell's interaction network and has great potential for hypothesis-driven research, which we plan to explore in future projects. Another future application consists in the spatial patterning of bacterial communities at microscale resolution to investigate microbial interactions.
Selected publications:
Gäbelein CG, Feng Q, Sarajlic E, Zambelli T, Guillaume-Gentil O, Kornmann B, Vorholt JA (2022) Mitochondria transplantation between living cells. PLoS Biol 20(3): e3001576. https://doi.org/10.1371/journal.pbio.3001576 [external pageAbstract]
Guillaume-Gentil O, Gäbelein CG, Schmieder S, Martinez V, Zambelli T, Künzler M, Vorholt JA (2022) Injection into and extraction from single fungal cells. Commun Biol. 2022 Mar 1;5(1):180. doi: 10.1038/s42003-022-03127-z. [external pageAbstract]
Mittelviefhaus M, Müller DB, Zambelli T, Vorholt JA (2019) A modular atomic force microscopy approach reveals a large range of hydrophobic adhesion forces of the leaf microbiota. ISME J, 13:1878-1882 [external pageAbstract]
Guillaume-Gentil O, Mittelviefhaus M, Dorwling-Carter L, Zambelli T, Vorholt JA (2018) FluidFM applications in single-cell biology. Open-Space Microfluidics: Concepts, implementations, applications John Wiley & Sons p. 325-355 [external pageAbstract]
Guillaume-Gentil O, Rey T, Kiefer P, Ibáñez A.J., Steinhoff R, Brönnimann R, Dorwling-Carter L, Zambelli T, Zenobi R, Vorholt JA (2017) Single-cell mass spectrometry of metabolites extracted from live cells by fluidic force microscopy. Anal Chem 89:5017-23 [external pageAbstract]
Guillaume-Gentil O, Grindberg RV, Kooger R, Dorwling-Cater L, Martinez V, Ossola D, Pilhofer M, Zambelli T, Vorholt JA (2016) Tunable single-cell extraction for molecular analysis. Cell 166: 506-516 external page[Abstract]
Potthoff E, Ossola D, Zambelli T, Vorholt JA (2015) Bacterial adhesion force quantification by fluidic force microscopy. Nanoscale 7:4070-4079 external page[Abstract]
Guillaume-Gentil O, Potthoff E, Franz C.M., Zambelli T, Vorholt JA (2014) Force-controlled manipulation of single cells: from AFM to FluidFM. Trends Biotech. 32:381-388 external page[Abstract]
Guillaume-Gentil O, Zambelli T, Vorholt JA (2014) Isolation of single mammalian cells from adherent cultures by fluidic force microscopy. Lab-on-a-Chip 14:402-414 external page[Abstract] http://www.ncbi.nlm.nih.gov/pubmed/?term=24270585)
Potthoff E, Franco D, D’Alessandro D, Starck C, Falk V, Zambelli T, Vorholt JA, Poulikakos D, Ferrari A (2014) Toward a rational design of surface textures promoting endothelialization. Nano Letters 14:1069-1079 external page[Abstract]
Guillaume-Gentil O, Potthoff E, Ossola D, Dörig P, Zambelli T, Vorholt JA (2013) Force-controlled fluidic injection into single cell nuclei. Small 9:1904-1907 external page[Abstract]
Stiefel P, Zambelli T, Vorholt JA (2013) Isolation of optically targeted single bacteria by application of fluidic force microscopy to aerobic anoxygenic phototrophs from the phyllosphere. Appl Environ Microbiol. 79:4895-4905 external page[Abstract]
Potthoff E, Guillaume-Gentil O, Ossola, D, Polesel-Maris J, LeibundGut-Landmann S, Zambelli T, Vorholt JA (2012) Rapid and serial quantification of adhesion forces of yeast and mammalian cells. PLoS One. 7:e52712 external page[Abstract]
Stiefel S, Schmidt F, Dörig P, Behr P, Zambelli T, Vorholt JA, Mercer J (2012) Cooperative vaccinia infection demonstrated at the single-cell level using FluidFM. Nano Letters. 12:4219-4227 external page[Abstract]
Dörig P, Stiefel P, Behr P, Sarajlic E, Bijl D, Gabi M, Vörös J, Vorholt JA, Zambelli T (2010) Force-controlled spatial manipulation of viable mammalian cells and microorganisms. Appl. Phys. Lett. 97:023701 external page[Abstract]