We've already highlighted reviews (Part I) and publications using FluidFM for nanoprinting (Part II) and cell adhesion measurements (Part III). Today, let's dive into publications using FluidFM technology for single-cell injection into and extraction from single cells!
Discover 2022 publications using FluidFM for single-cell injection and extraction!
Extracting, injecting and transplanting mitochondria in living single cells
Christoph Gäbelein and colleagues from the lab of Julia Vorholt develop a FluidFM-based approach to extract, inject, and transplant organelles from and into living cells with subcellular spatial resolution. The authors show how the use of FluidFM overcomes existing limitations in organelle manipulation, specifically that extraction and injection of varying amounts of organelles from single cells, which had not been possible before, as well as exploring the response of single cells to transferred mitochondria. Thanks to the gentle mode of probe insertion and extraction of the FluidFM, this approach ensures efficient and scalable organelle manipulation while maintaining cell viability. This approach opens new prospects for the study of organelle physiology and homeostasis, but also for therapy, mechanobiology, and synthetic biology.
Schematic of organelle extraction and injection using FluidFM, showing (A) Extraction where volumes are tuned by applying negative pressure (−Δp); B) Selective extraction of organelle components by tuning the aperture size and thus the applicable range of applied fluidic forces. (C) Mitochondria injection into single cells by applying positive pressure (+Δp) once the cantilever was inserted into the recipient cell. Figure taken from Gäbelein et al PLoS Biol 2022 (License: CC BY 4.0)
Injecting into and extracting from fungal cells without having to degrade the cell wall
In this study, Orane Guillaume-Gentil and colleagues from the lab of Julia Vorholt show that FluidFM technology enables injection of solutions and cytoplasmic fluid extraction into and out of individual fungal cells. This proof-of-concept study demonstrates that the use of FluidFM allows traversing the cell wall of living fungi with the nano-fluidic probe, which removes the need to generate protoplasts by degrading the cell wall, for subsequent delivery into or extraction from individual fungal cells. This approach opens new opportunities for manipulating and analyzing fungi, from unicellular model yeasts to multicellular filamentous fungi.
Illustration and representative images of the Live-seq sampling procedure using FluidFM (here, applied on brown preadipocyte IBA cells). The white arrows indicate the application of under- or overpressure. The black arrows indicate the amount of buffer and extract in the probe. Figure adapted from Chen et al Nature 2022 (License: CC BY 4.0)
Part IV: injection/extraction with FluidFM