Fluorescence fluctuations analysis in nanoapertures: physical concepts and biological applications

Abstract

During the past years, nanophotonics has provided new approaches to study the biological processes below the optical diVraction limit. How single molecules diVuse, bind and assemble can be studied now at the nanometric level, not only in solutions but also in complex and crowded environments such as in live cells. In this context Xuorescence Xuctuations spectroscopy is a unique tool since it has proven to be easy to use in combination with nanostructures, which are able to conWne light in nanometric volumes. We review here recent advances in Xuorescence Xuctuations' analysis below the optical diVraction limit with a special focus on nanoapertures milled in metallic Wlms. We discuss applications in the Weld of single-molecule detection, DNA sequencing and membrane organization, and underscore some potential perspectives of this new emerging technology.

Enhanced fluorescence from metal nanoapertures: physical characterizations and biophotonic applications

“…These applications are reviewed for instance in. 15 Briefly, the illuminated area well below the diffraction limit has proven of essential interest for FCS studies at high (micromolar) molecular concentrations. 3 It also offers an elegant way to probe an area well below the diffraction limit in lipid bilayers 16,17 as well as in live cell membranes.…”

Extending the knowledge in histochemistry and cell biology

“…In a fascinating overview, Lenne described his recent approaches investigating cell-surface dynamics and mechanics (Lenne 2009;Lenne et al 2008). By using FCS together with nanometric apertures, the resolution limit of molecular diVusion signals was enhanced and signals intensiWed.…”

“…microfabricated nanoapertures) can not only greatly improve already established techniques (e.g. FCS) but also lead the pave for future technologies such as single molecule DNA sequencing Lenne et al 2008).…”

State-of-the-art technologies, current opinions and developments, and novel findings: news from the field of histochemistry and cell biology

“…A newly emerging technology in the Weld of single-molecule detection, DNA sequencing and membrane organization, namely, Xuorescence Xuctuations analysis in nanoapertures, was the topic of a review by Lenne and colleagues ("Fluorescence Xuctuations analysis in nanoapertures: physical concepts and biological applications"; Lenne et al 2008). The authors reviewed literature on dynamic measurements using nanostructures such as nanoapertures milled in metallic Wlms, which can conWne light in volumes at least three orders of magnitude smaller than achievable with confocal microscopy, providing small observation volumes and, sometimes, enhanced Xuorescence detection.…”