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. (Graph Presented).
| Original language | English (US) |
|---|---|
| Pages (from-to) | 8122-8130 |
| Number of pages | 9 |
| Journal | ACS Nano |
| Volume | 9 |
| Issue number | 8 |
| DOIs | |
| State | Published - Aug 25 2015 |
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Keywords
- direct stochastic optical reconstruction microscopy (dSTORM)
- nanopipette
- quantitative super-resolution microscopy
- scanning ion conductance microscopy (SICM)
- single-molecule deposition
ASJC Scopus subject areas
- Engineering(all)
- Materials Science(all)
- Physics and Astronomy(all)
Cite this
Quantitative Super-Resolution Microscopy of Nanopipette-Deposited Fluorescent Patterns. / Hennig, Simon; Van De Linde, Sebastian; Bergmann, Stephan; Huser, Thomas R; Sauer, Markus.
In: ACS Nano, Vol. 9, No. 8, 25.08.2015, p. 8122-8130.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Quantitative Super-Resolution Microscopy of Nanopipette-Deposited Fluorescent Patterns
AU - Hennig, Simon
AU - Van De Linde, Sebastian
AU - Bergmann, Stephan
AU - Huser, Thomas R
AU - Sauer, Markus
PY - 2015/8/25
Y1 - 2015/8/25
N2 - 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. (Graph Presented).
AB - 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. (Graph Presented).
KW - direct stochastic optical reconstruction microscopy (dSTORM)
KW - nanopipette
KW - quantitative super-resolution microscopy
KW - scanning ion conductance microscopy (SICM)
KW - single-molecule deposition
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UR - http://www.scopus.com/inward/citedby.url?scp=84940118398&partnerID=8YFLogxK
U2 - 10.1021/acsnano.5b02220
DO - 10.1021/acsnano.5b02220
M3 - Article
C2 - 26173009
AN - SCOPUS:84940118398
VL - 9
SP - 8122
EP - 8130
JO - ACS Nano
JF - ACS Nano
SN - 1936-0851
IS - 8
ER -