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Quantitative Chemical Delivery of Quantum Dots into the Cytosol of Cells

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Quantum Dots

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2135))

Abstract

The ability to image single molecules in living cells has been impaired by the absence of bright, photostable fluorophores. Quantum dots (QDs) offer an attractive solution to this problem due to their exceptional photostability and brightness. Here, we describe in detail a protocol to chemically deliver functionalized QDs into the cytosol of living cells, based on cell-penetrating poly(disulfide)s (CPDs). This protocol is highly efficient and delivers hundreds of QDs per cell after incubation of cells with functionalized QDs at nanomolar concentrations. We also detail a pipeline for automated detection and tracking of diffusive QDs in living cells, which may provide a useful means to study the biophysical properties of the cytosol and their dynamics. Last, we describe a protocol for conjugating streptavidin fusion proteins to QDs, in order to permit the codelivery of QDs with functional proteins of interest into cells. The protocol has been successfully applied to a broad range of different cell types, thus offering a flexible and generalizable means to image single molecules in living cells.

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Acknowledgments

This work was supported by the Medical Research Council (file reference number MC_UP_1201/13), HFSP (CDA00034/2017-C) for E.D., a Michael Neuberger Studentship for J.L.W, and the Astra/Zeneca Blue Skies Initiative for A.S.

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Authors and Affiliations

  1. MRC Laboratory of Molecular Biology, Cambridge, UK

    Joseph L. Watson, Alessandra Stangherlin & Emmanuel Derivery

Authors
  1. Joseph L. Watson
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  2. Alessandra Stangherlin
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  3. Emmanuel Derivery
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Corresponding author

Correspondence to Emmanuel Derivery .

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Editors and Affiliations

  1. Biomedical Nanotechnology Group, Federal University of Pernambuco, Recife, Pernambuco, Brazil

    Adriana Fontes

  2. Biomedical Nanotechnology Group, Federal University of Pernambuco, Recife, Pernambuco, Brazil

    Beate S. Santos

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Watson, J.L., Stangherlin, A., Derivery, E. (2020). Quantitative Chemical Delivery of Quantum Dots into the Cytosol of Cells. In: Fontes, A., Santos, B. (eds) Quantum Dots. Methods in Molecular Biology, vol 2135. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0463-2_10

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  • DOI: https://doi.org/10.1007/978-1-0716-0463-2_10

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0462-5

  • Online ISBN: 978-1-0716-0463-2

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