Nanodroplet-Based Reagent Delivery into Water-in-Fluorinated-Oil Droplets

Bo Zhu, Zhe Du, Yancen Dai, Tetsuya Kitaguchi, Sebastian Behrens, Burckhard Seelig

Research output: Contribution to journalArticlepeer-review


In vitro compartmentalization (IVC) is a technique for generating water-in-oil microdroplets to establish the genotype (DNA information)–phenotype (biomolecule function) linkage required by many biological applications. Recently, fluorinated oils have become more widely used for making microdroplets due to their better biocompatibility. However, it is difficult to perform multi-step reactions requiring the addition of reagents in water-in-fluorinated-oil microdroplets. On-chip droplet manipulation is usually used for such purposes, but it may encounter some technical issues such as low throughput or time delay of reagent delivery into different microdroplets. Hence, to overcome the above issues, we demonstrated a nanodroplet-based approach for the delivery of copper ions and middle-sized peptide molecules (human p53 peptide, 2 kDa). We confirmed the ion delivery by microscopic inspection of crystal formation inside the microdroplet, and confirmed the peptide delivery using a fluorescent immunosensor. We believe that this nanodroplet-based delivery method is a promising approach to achieving precise control for a broad range of fluorocarbon IVC-based biological applications, including molecular evolution, cell factory engineering, digital nucleic acid detection, or drug screening.

Original languageEnglish (US)
Article number768
Issue number8
StatePublished - Aug 2023

Bibliographical note

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© 2023 by the authors.


  • biosensor
  • microdroplet
  • microfluidic device
  • nanodroplet
  • reagent delivery


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