Journal
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
Volume 106, Issue 42, Pages 17735-17740Publisher
NATL ACAD SCIENCES
DOI: 10.1073/pnas.0907367106
Keywords
caspase; live cell imaging; plasmonic nanoparticles; protease sensor; single-molecule imaging
Categories
Funding
- National Institutes of Health [R01-GM77856, NOT-OD-09-056, CA72006, CA128765]
- US Air Force Office of Scientific Research-Korea Ministry of Education-Science and Technology Nano-Bio-Information Technology Program [K20716000001-07A0400-00100]
- Lawrence Berkeley National Laboratory [LB08003826]
- Leukemia and Lymphoma Society Fellowship [5552-06]
- National Institute of General Medical Sciences [1 R25 GM56847]
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The use of plasmon coupling in metal nanoparticles has shown great potential for the optical characterization of many biological processes. Recently, we have demonstrated the use of plasmon rulers'' to observe conformational changes of single biomolecules in vitro. Plasmon rulers provide robust signals without photo-bleaching or blinking. Here, we show the first application of plasmon rulers to in vivo studies to observe very long trajectories of single biomolecules in live cells. We present a unique type of plasmon ruler comprised of peptide-linked gold nanoparticle satellites around a core particle, which was used as a probe to optically follow cell-signaling pathways in vivo at the single-molecule level. These crown nanoparticle plasmon rulers'' allowed us to continuously monitor trajectories of caspase-3 activity in live cells for over 2 h, providing sufficient time to observe early-stage caspase-3 activation, which was not possible by conventional ensemble analyses.
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