ABSTRACT
Tracking dynamic microtubule ends in fluorescence microscopy movies provides insight into the statistical properties of microtubule dynamics and is vital for further analysis that requires knowledge of the trajectories of the microtubule ends. Here we analyse the performance of a previously developed automated microtubule end tracking routine; this has been optimized for comparatively low signal-to-noise image sequences that are characteristic of microscopy movies of dynamic microtubules growing in vitro. Sequences of simulated microtubule images were generated assuming a variety of different experimental conditions. The simulated movies were then tracked and the tracking errors were characterized. We found that the growth characteristics of the microtubules within realistic ranges had a negligible effect on the tracking precision. The fluorophore labelling density, the pixel size of the images, and the exposure times were found to be important parameters limiting the tracking precision which could be explained using concepts of single molecule localization microscopy. The signal-to-noise ratio was found to be a good single predictor of the tracking precision: typical experimental signal-to-noise ratios lead to tracking precisions in the range of tens of nanometres, making the tracking program described here a useful tool for dynamic microtubule end tracking with close to molecular precision.
Subject(s)
Image Processing, Computer-Assisted/methods , Microscopy, Fluorescence/methods , Microtubules/ultrastructure , Algorithms , Computer Simulation , Fluorescent DyesABSTRACT
We have investigated the effects of tuning the localized surface plasmon resonances (LSPRs) of silver nanoparticles on the fluorescence intensity, lifetime, and Raman signal from nearby fluorophores. The presence of a metallic structure can alter the optical properties of a molecule by increasing the excitation field, and by modifying radiative and nonradiative decay mechanisms. By careful choice of experimental parameters we have been able to decouple these effects. We observe a fourfold increase in fluorescence enhancement and an almost 30-fold increase in decay rate from arrays of Ag nanoparticles, when the LSPR is tuned to the emission wavelength of a locally situated fluorophore. This is consistent with a greatly increased efficiency for energy transfer from fluorophores to surface plasmons, resulting in a significant increase in quantum yield. Additionally, spatial mapping of the surface enhanced Raman scattering signal from a nanoparticle array reveals highly localized differences in the excitation field.
Subject(s)
Metal Nanoparticles/chemistry , Spectrum Analysis, Raman/methods , Surface Plasmon Resonance/methods , Microscopy, Atomic ForceABSTRACT
We demonstrate lasing action with a high spontaneous emission factor and temperature insensitivity in InAs/InGaAs quantum dots (QD) embedded in photonic crystal nanocavities. A quality factor (Q) of over 10,000 was achieved by suppressing the material absorption by QDs uncoupled to the cavity mode. High Q cavities exhibited ultra low threshold lasing with a spontaneous emission factor of 0.7. Less frequent carrier escape from the QDs, which was primarily favored by high potential barrier energy, enabled low threshold lasing up to 90 K.
Subject(s)
Arsenicals/chemistry , Gallium/chemistry , Indium/chemistry , Lasers, Semiconductor , Nanotechnology/instrumentation , Quantum Dots , Equipment Design , Equipment Failure Analysis , Photons , Quality Control , TemperatureABSTRACT
Modulation-doped GaAs v-groove quantum wires (QWRs) have been fabricated with novel electrical contacts made to two-dimensional electron-gas (2DEG) reservoirs. Here, we present longitudinal photocurrent (photoconductivity/PC) spectroscopy measurements of a single QWR. We clearly observe conductance in the ground-state one-dimensional subbands; in addition, a highly temperature-dependent response is seen from other structures within the v-groove. The latter phenomenon is attributed to the effects of structural topography and localization on carrier relaxation. The results of power-dependent PC measurements suggest that the QWR behaves as a series of weakly interacting localized states, at low temperatures.
