Transfection of living HeLa cells with fluorescent poly-cytosine encapsulated Ag nanoclusters

The fluorescence of silver clusters encapsulated by single stranded oligo-DNA (24 cytosine base pairs, C(24):Ag(n)) was used to monitor the transfection of this new silver/DNA fluorophore inside living HeLa cells. For this, the C(24):Ag(n) molecules were complexed with a commercially available transfection reagent Lipofectamine and the internalization of C(24):Ag(n) was followed with confocal fluorescence microscopy. Bright near-infrared fluorescence was observed from inside the transfected HeLa cells, when exciting with 633 nm excitation, opening up the possibility for the use of these C(24):Ag(n) clusters for biological labelling and imaging of living cells and for monitoring the transfection process with limited harm to the living cells.

[1]  S. Hell Far-Field Optical Nanoscopy , 2007, Science.

[2]  J. Lippincott-Schwartz,et al.  Imaging Intracellular Fluorescent Proteins at Nanometer Resolution , 2006, Science.

[3]  D. Crothers,et al.  Solution structural studies of the Ag(I)-DNA complex. , 1981, Nucleic acids research.

[4]  G. Mocz Fluorescent Proteins and Their Use in Marine Biosciences, Biotechnology, and Proteomics , 2007, Marine Biotechnology.

[5]  P. Leder,et al.  Splicing and the formation of stable RNA , 1979, Cell.

[6]  Shuming Nie,et al.  Engineering Luminescent Quantum Dots for In Vivo Molecular and Cellular Imaging , 2006, Annals of Biomedical Engineering.

[7]  R. Dickson,et al.  Water-soluble Ag nanoclusters exhibit strong two-photon-induced fluorescence. , 2008, Journal of the American Chemical Society.

[8]  J. Hofkens,et al.  Photophysics of a water-soluble rylene dye: Comparison with other fluorescent molecules for biological applications , 2004 .

[9]  D. Balding,et al.  HLA Sequence Polymorphism and the Origin of Humans , 2006 .

[10]  R. Dickson,et al.  Shuttle-based fluorogenic silver-cluster biolabels. , 2009, Angewandte Chemie.

[11]  Robin H. A. Ras,et al.  Color tunability and electrochemiluminescence of silver nanoclusters. , 2009, Angewandte Chemie.

[12]  Tom Vosch,et al.  Oligonucleotide-stabilized Ag nanocluster fluorophores. , 2008, Journal of the American Chemical Society.

[13]  S. Pal,et al.  Structural and Functional Characterization of Luminescent Silver−Protein Nanobioconjugates , 2008 .

[14]  Robert M Dickson,et al.  DNA-templated Ag nanocluster formation. , 2004, Journal of the American Chemical Society.

[15]  M. Roeffaers,et al.  Characterization of fluorescence in heat-treated silver-exchanged zeolites. , 2009, Journal of the American Chemical Society.

[16]  H. Luhmann,et al.  Novel fluorescent core-shell nanocontainers for cell membrane transport. , 2008, Biomacromolecules.

[17]  P. Hofschneider,et al.  Appearance of β-lactamase activity in animal cells upon liposome-mediated gene transfer , 1980 .

[18]  S. Hell,et al.  Fluorescence nanoscopy by ground-state depletion and single-molecule return , 2008, Nature Methods.

[19]  M. Zimmer GFP: from jellyfish to the Nobel prize and beyond. , 2009, Chemical Society reviews.

[20]  Moungi G Bawendi,et al.  Compact biocompatible quantum dots functionalized for cellular imaging. , 2008, Journal of the American Chemical Society.

[21]  Eugenia Kumacheva,et al.  Photogeneration of Fluorescent Silver Nanoclusters in Polymer Microgels , 2005 .

[22]  Shaojun Dong,et al.  Sensitive detection of cysteine based on fluorescent silver clusters. , 2009, Biosensors & bioelectronics.

[23]  M. Roeffaers,et al.  Photoactivation of silver-exchanged zeolite A. , 2008, Angewandte Chemie.

[24]  Clive R. Bagshaw,et al.  The photochemistry of fluorescent proteins: implications for their biological applications. , 2009, Chemical Society reviews.

[25]  Hideki Nabika,et al.  Enhanced Emission from Photoactivated Silver Clusters Coupled with Localized Surface Plasmon Resonance , 2009 .

[26]  S. Arya,et al.  Optical rotatory dispersion and circular dichroism of silver(I):Polyribonucleotide complexes , 1975 .

[27]  S. Gambhir,et al.  Quantum Dots for Live Cells, in Vivo Imaging, and Diagnostics , 2005, Science.

[28]  P. Goodwin,et al.  Base-Directed Formation of Fluorescent Silver Clusters. , 2008, The journal of physical chemistry. C, Nanomaterials and interfaces.

[29]  R. Dickson,et al.  Ag Nanocluster Formation Using a Cytosine Oligonucleotide Template. , 2007, The journal of physical chemistry. C, Nanomaterials and interfaces.

[30]  E. Wang,et al.  Oligonucleotide-stabilized Ag nanoclusters as novel fluorescence probes for the highly selective and sensitive detection of the Hg2+ ion. , 2009, Chemical communications.

[31]  K. Braeckmans,et al.  Towards a better understanding of the dissociation behavior of liposome-oligonucleotide complexes in the cytosol of cells. , 2005, Journal of controlled release : official journal of the Controlled Release Society.

[32]  C. Blum,et al.  Single-molecule spectroscopy of fluorescent proteins , 2009, Analytical and bioanalytical chemistry.

[33]  S. Bhatia,et al.  Probing the Cytotoxicity Of Semiconductor Quantum Dots. , 2004, Nano letters.

[34]  E. Neumann,et al.  Gene transfer into mouse lyoma cells by electroporation in high electric fields. , 1982, The EMBO journal.

[35]  E. Gwinn,et al.  Sequence‐Dependent Fluorescence of DNA‐Hosted Silver Nanoclusters , 2008 .

[36]  A. Hirko,et al.  Cationic lipid vectors for plasmid DNA delivery. , 2003, Current medicinal chemistry.

[37]  Igor L. Medintz,et al.  Quantum dot bioconjugates for imaging, labelling and sensing , 2005, Nature materials.

[38]  N. Yang,et al.  Cytokine gene therapy of cancer using gene gun technology: superior antitumor activity of interleukin-12. , 1997, Human gene therapy.

[39]  R. Tsien,et al.  The Fluorescent Toolbox for Assessing Protein Location and Function , 2006, Science.

[40]  J. Perry,et al.  Electron transfer-induced blinking in Ag nanodot fluorescence. , 2009, The journal of physical chemistry. C, Nanomaterials and interfaces.

[41]  R. Dickson,et al.  In vitro and intracellular production of peptide-encapsulated fluorescent silver nanoclusters. , 2007, Angewandte Chemie.

[42]  R. Dickson,et al.  Photoactivated fluorescence from individual silver nanoclusters. , 2001, Science.

[43]  Mario R. Capecchi,et al.  High efficiency transformation by direct microinjection of DNA into cultured mammalian cells , 1980, Cell.

[44]  K Dane Wittrup,et al.  Monovalent, reduced-size quantum dots for imaging receptors on living cells , 2008, Nature Methods.

[45]  R. Mulligan,et al.  Synthesis of rabbit β-globin in cultured monkey kidney cells following infection with a SV40 β-globin recombinant genome , 1979, Nature.

[46]  Michael W. Davidson,et al.  The fluorescent protein palette: tools for cellular imaging. , 2009, Chemical Society reviews.

[47]  Michael D. Mason,et al.  Ultra-high resolution imaging by fluorescence photoactivation localization microscopy. , 2006, Biophysical journal.

[48]  C. Seidel,et al.  Photobleaching of Fluorescent Dyes under Conditions Used for Single-Molecule Detection:  Evidence of Two-Step Photolysis. , 1998, Analytical chemistry.

[49]  Peter Dedecker,et al.  A stroboscopic approach for fast photoactivation-localization microscopy with Dronpa mutants. , 2007, Journal of the American Chemical Society.

[50]  U. Kubitscheck,et al.  Photostability Data for Fluorescent Dyes: An Update. , 2002 .

[51]  J Michiels,et al.  Identification of different emitting species in the red fluorescent protein DsRed by means of ensemble and single-molecule spectroscopy , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[52]  Tom Vosch,et al.  Strongly emissive individual DNA-encapsulated Ag nanoclusters as single-molecule fluorophores , 2007, Proceedings of the National Academy of Sciences.

[53]  R. Dickson,et al.  Live Cell Surface Labeling with Fluorescent Ag Nanocluster Conjugates † , 2008, Photochemistry and photobiology.

[54]  T. Pradeep,et al.  PAPER www.rsc.org/materials | Journal of Materials Chemistry Interfacial synthesis of luminescent 7 kDa silver clusters†‡ , 2009 .

[55]  Junhua Yu,et al.  Optically modulated fluorophores for selective fluorescence signal recovery. , 2009, Journal of the American Chemical Society.

[56]  N. Makarava,et al.  Water-soluble hybrid nanoclusters with extra bright and photostable emissions: a new tool for biological imaging. , 2005, Biophysical journal.

[57]  M. Ameloot,et al.  Visualization of membrane rafts using a perylene monoimide derivative and fluorescence lifetime imaging. , 2007, Biophysical journal.

[58]  Vincent Noireaux,et al.  In Vivo Imaging of Quantum Dots Encapsulated in Phospholipid Micelles , 2002, Science.

[59]  F. Marshall,et al.  In vivo molecular and cellular imaging with quantum dots. , 2005, Current opinion in biotechnology.

[60]  E. Gwinn,et al.  Hairpins with Poly-C Loops Stabilize Four Types of Fluorescent Agn:DNA , 2009 .

[61]  B. Lounis,et al.  Fluorescent silver oligomeric clusters and colloidal particles , 2005 .

[62]  Mark Van der Auweraer,et al.  New picosecond laser system for easy tunability over the whole ultraviolet/visible/near infrared wavelength range based on flexible harmonic generation and optical parametric oscillation , 2001 .