Quantitative super-resolution microscopy of nanopipette-deposited fluorescent patterns

Simon Hennig, Sebastian van de Linde, Stephan Bergmann, Thomas Huser, Markus Sauer

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

We describe a method for the deposition of minute amounts of fluorophore-labeled oligonucleotides with high local precision in conductive and transparent solid layers of poly(vinyl alcohol) (PVA) doped with glycerin and cysteamine (PVA-G-C layers). Deposition of negatively charged fluorescent molecules was accomplished with a setup based on a scanning ion conductance microscope (SICM) using nanopipettes with tip diameters of ∼100 nm by using the ion flux flowing between two electrodes through the nanopipette. To investigate the precision of the local deposition process, we performed in situ super-resolution microscopy by direct stochastic optical reconstruction microscopy (dSTORM). Exploiting the single-molecule sensitivity and reliability of dSTORM, we determine the number of fluorescent molecules deposited in single spots. The correlation of applied charge and number of deposited molecules enables the quantification of delivered molecules by measuring the charge during the delivery process. We demonstrate the reproducible deposition of 3-168 fluorescent molecules in single spots and the creation of fluorescent structures. The fluorescent structures are highly stable and can be reused several times.

Original languageEnglish
Pages (from-to)8122-8130
Number of pages9
JournalACS Nano
Volume9
Issue number8
Early online date14 Jul 2015
DOIs
Publication statusPublished - 25 Aug 2015
Externally publishedYes

Keywords

  • dSTORM
  • nanopipette
  • super-resolution microscopy
  • SICM
  • single-molecule deposition

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