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Microscopy-Based High-Throughput Analysis of Cells Interacting with Nanostructures

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Systems Biology Application in Synthetic Biology

Abstract

Due to their small size and related interesting properties, artificial nanomaterials are utilized for a great number of biological and medical applications. Cell entry routes, intracellular trafficking and processing of nanoparticles, which determine their fate, efficiency, and toxicity, are depending on various parameters of the specific nanomaterial, such as size, surface charge, surface chemistry and elasticity. Nanoparticle-cell interactions are typically elucidated by means of fluorescence microscopy as cell functions can be observed by a multiplicity of commercially available probes. For the quantification of cell features from images (image cytometry), computer-based algorithms are favoured to avoid bias introduced by the subjective perception of the observer. By applying high throughput microscopy in combination with digital image cytometry the screening of high numbers of cells is made possible yielding statistically meaningful results. In this chapter methods from digital image cytometry are described for assessing the interactions of cells with nanostructures.

This chapter is adopted from the PhD thesis of Dr. Raimo Hartmann, submitted and accepted by the Physics Department of the Philipps Universität Marburg in 2015.

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Notes

  1. 1.

    Also referred to as high-content screening (HCS).

  2. 2.

    d h = hydrodynamic diameter.

  3. 3.

    1321 N1, SH-SY5Y, Raw267.4, A549, hCMEC, HepG2, and HEK293 cells.

  4. 4.

    Jan et al. [52] did not provide any further characterization in their work.

  5. 5.

    Depending on the optical properties of the fluorescent complex and the instrumentation.

  6. 6.

    Splines are piecewise-defined polynomial functions.

  7. 7.

    Voronoi diagrams describe a distance-controlled partitioning of a plane into regions based on seeds, cf. Figure 9.3e [73].

  8. 8.

    For instance, in the case of adherent cells, no detachment and transfer into certain buffers prior to cytometric measurements are required.

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  1. Fachbereich Physik, Philipps Universität Marburg, Marburg, Germany

    Raimo Hartmann & Wolfgang J. Parak

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  1. Raimo Hartmann
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Correspondence to Raimo Hartmann .

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  1. Computational and Systems Biology Lab, National Centre for Cell Science, Pune, India

    Shailza Singh

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Hartmann, R., Parak, W.J. (2016). Microscopy-Based High-Throughput Analysis of Cells Interacting with Nanostructures. In: Singh, S. (eds) Systems Biology Application in Synthetic Biology. Springer, New Delhi. https://doi.org/10.1007/978-81-322-2809-7_9

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