The Auger recombination coefficient in quasi-bulk InxGa1−xN (x∼9%–15%) layers grown on GaN (0001) is measured by a photoluminescence technique. The samples vary in InN composition, thickness, and threading dislocation density. Throughout this sample set, the measured Auger coefficient ranges from 1.4×10−30to2.0×10−30cm6s−1. The authors argue that an Auger coefficient of this magnitude, combined with the high carrier densities reached in blue and green InGaN∕GaN (0001) quantum well light-emitting diodes (LEDs), is the reason why the maximum external quantum efficiency in these devices is observed at very low current densities. Thus, Auger recombination is the primary nonradiative path for carriers at typical LED operating currents and is the reason behind the drop in efficiency with increasing current even under room-temperature (short-pulsed, low-duty-factor) injection conditions.

Auger recombination in InGaN measured by photoluminescence

Nitride-based light-emitting diodes (LEDs) suffer from a reduction (droop) of the internal quantum efficiency with increasing injection current. This droop phenomenon is currently the subject of intense research worldwide, as it delays general lighting applications of GaN-based LEDs. Several explanations of the efficiency droop have been proposed in recent years, but none is widely accepted. This feature article provides a snapshot of the present state of droop research, reviews currently discussed droop mechanisms, contextualizes them, and proposes a simple yet unified model for the LED efficiency droop.

Illustration of LED efficiency droop (details in Fig. 13).

Efficiency droop in nitride-based light-emitting diodes

A major challenge for fluorescence imaging of living mammalian cells is maintaining viability following prolonged exposure to excitation illumination. We have monitored the dynamics of mitochondrial distribution in hamster embryos at frequent intervals over 24 h using two-photon microscopy (1,047 nm) while maintaining blastocyst, and even fetal, developmental competence. In contrast, confocal imaging for only 8 h inhibits development, even without fluorophore excitation. Photo-induced production of H2O2 may account, in part, for this inhibition. Thus, two-photon microscopy, but not confocal microscopy, has permitted long-term fluorescence observations of the dynamics of three-dimensional cytoarchitecture in highly photosensitive specimens such as mammalian embryos.

Long-term two-photon fluorescence imaging of mammalian embryos without compromising viability

Fluorescence spectroscopy is widely used in biological research. Until recently, essentially all fluorescence experiments were performed using optical energy which has radiated to the far-field. By far-field we mean at least several wavelengths from the fluorophore, but propagating far-field radiation is usually detected at larger macroscopic distances from the sample. In recent years there has been a growing interest in the interactions of fluorophores with metallic surfaces or particles. Near-field interactions are those occurring within a wavelength distance of an excited fluorophore. The spectral properties of fluorophores can be dramatically altered by near-field interactions with the electron clouds present in metals. These interactions modify the emission in ways not seen in classical fluorescence experiments. In this review we provide an intuitive description of the complex physics of plasmons and near-field interactions. Additionally, we summarize the recent work on metal-fluorophore interactions and suggest how these effects will result in new classes of experimental procedures, novel probes, bioassays and devices.

Plasmon-controlled fluorescence: a new paradigm in fluorescence spectroscopy

Auger recombination is determined to be the limiting factor for quantum efficiency for InGaN–GaN (0001) light-emitting diodes (LEDs) at high current density. High-power double-heterostructure (DH) LEDs are grown by metal-organic chemical vapor deposition. By increasing the active layer thickness, DH LEDs can reach a maximum in quantum efficiency at current densities above 200A∕cm2. Encapsulated thin-film flip-chip DH LEDs with peak wavelength of 432nm have an external quantum efficiency of 40% and output power of 2.3W at 2A.

Blue-emitting InGaN–GaN double-heterostructure light-emitting diodes reaching maximum quantum efficiency above 200A∕cm2

Pulsed light–current characteristics of InGaN/GaN quantum welllight-emitting diodes have been measured as a function of temperature, with sublinear behavior observed over the whole temperature range, 130–330 K. A distinctive temperature dependence is also noted where the light output, at a fixed current, initially increases with temperature, before reaching a maximum at 250 K and then decreases with subsequent increases in temperature. On the basis of a drift diffusion model, we can explain the sublinear light–current characteristics and the temperature dependence by the influence of the large acceptor ionization energy in Mg-doped GaN together with a triangular density of states function characteristic of localized states. Without the incorporation of localization effects, we are unable to reproduce the temperature dependence whilst maintaining emission at the observed wavelength. This highlights the importance of localization effects on device performance.

/pdf/carrier-leakage-in-ingan-quantum-well-light-emitting-diodes-423ge7zggy.pdf

Carrier leakage in InGaN quantum well light-emitting diodes emitting at 480 nm

In many studies, the value of the experimentally determined internal piezoelectric field has been reported to be significantly smaller than theoretical values. We believe this is due to an inappropriate approximation for the electric field within the depletion region, which is used in the analysis of experimental data, and we propose an alternative method. Using this alternative, we have measured the strength of the internal field of InGaN p-i-n structures, using reverse bias photocurrent absorption spectroscopy and by fitting the bias dependent peak energy using microscopic theory based on the screened Hartree-Fock approximation. The results agree with those using material constants interpolated from binary values.

/pdf/determination-of-the-piezoelectric-field-in-ingan-quantum-4uigzl1rvk.pdf

Determination of the piezoelectric field in InGaN quantum wells

Mammalian oocytes contain lipid droplets that are a store of fatty acids, whose metabolism plays a substantial role in pre-implantation development. Fluorescent staining has previously been used to image lipid droplets in mammalian oocytes and embryos, but this method is not quantitative and often incompatible with live cell imaging and subsequent development. Here we have applied chemically specific, label-free coherent anti-Stokes Raman scattering (CARS) microscopy to mouse oocytes and pre-implantation embryos. We show that CARS imaging can quantify the size, number and spatial distribution of lipid droplets in living mouse oocytes and embryos up to the blastocyst stage. Notably, it can be used in a way that does not compromise oocyte maturation or embryo development. We have also correlated CARS with two-photon fluorescence microscopy simultaneously acquired using fluorescent lipid probes on fixed samples, and found only a partial degree of correlation, depending on the lipid probe, clearly exemplifying the limitation of lipid labelling. In addition, we show that differences in the chemical composition of lipid droplets in living oocytes matured in media supplemented with different saturated and unsaturated fatty acids can be detected using CARS hyperspectral imaging. These results demonstrate that CARS microscopy provides a novel non-invasive method of quantifying lipid content, type and spatial distribution with sub-micron resolution in living mammalian oocytes and embryos.

/pdf/quantitative-imaging-of-lipids-in-live-mouse-oocytes-and-1zqrhl739n.pdf

Quantitative imaging of lipids in live mouse oocytes and early embryos using CARS microscopy

Nanoparticles have attracted enormous attention for biomedical applications as optical labels, drug-delivery vehicles and contrast agents in vivo. In the quest for superior photostability and biocompatibility, nanodiamonds are considered one of the best choices due to their unique structural, chemical, mechanical and optical properties. So far, mainly fluorescent nanodiamonds have been utilized for cell imaging. However, their use is limited by the efficiency and costs in reliably producing fluorescent defect centres with stable optical properties. Here, we show that single non-fluorescing nanodiamonds exhibit strong coherent anti-Stokes Raman scattering (CARS) at the sp3 vibrational resonance of diamond. Using correlative light and electron microscopy, the relationship between CARS signal strength and nanodiamond size is quantified. The calibrated CARS signal in turn enables the analysis of the number and size of nanodiamonds internalized in living cells in situ, which opens the exciting prospect of following complex cellular trafficking pathways quantitatively.

/pdf/coherent-anti-stokes-raman-scattering-microscopy-of-single-qt4cp1ko6i.pdf

Coherent anti-Stokes Raman scattering microscopy of single nanodiamonds

We have developed a multimodal multiphoton laser-scanning microscope for cell imaging featuring simultaneous acquisition of differential Coherent Antistokes Raman Scattering (D-CARS), two-photon fluorescence (TPF) and second harmonic generation (SHG) using a single 5 fs Ti:Sa broadband (660-970 nm) laser The spectral and temporal pulse requirements of these modalities were optimized independently by splitting the laser spectrum into three parts: TPF/SHG excitation (> 900 nm), CARS Pump excitation (< 730 nm), and CARS Stokes excitation (730-900 nm) In particular, by applying an equal linear chirp to pump and Stokes pulses using glass dispersion we achieved a CARS spectral resolution of 10 cm(-1), and acquired CARS images over the 1200-3800 cm(-1) vibrational range selected by the time delay between pump and Stokes A prism pulse compressor in the TPF/SHG excitation was used to achieve Fourier limited 30 fs pulses at the sample for optimum TPF and SHG D-CARS was implemented with few passive optical elements and enabled simultaneous excitation and detection of two vibrational frequencies with a separation adjustable from 20 cm(-1) to 150 cm(-1) for selective chemical contrast and background suppression The excitation/detection set-up using beam-scanning was built around a commercial inverted microscope stand providing conventional bright-field, differential interference contrast and epi-fluorescence for user-friendly characterization of biological samples Examples of CARS hyperspectral images and simultaneous acquisition of D-CARS, TPF and SHG images in both forward and epi-direction are shown on HeLa cells, stem-cell derived human adipocytes and mouse tissues

Iestyn Pope

Papers

Carrier leakage in InGaN quantum well light-emitting diodes emitting at 480 nm

Determination of the piezoelectric field in InGaN quantum wells

Quantitative imaging of lipids in live mouse oocytes and early embryos using CARS microscopy

Coherent anti-Stokes Raman scattering microscopy of single nanodiamonds

Simultaneous hyperspectral differential-CARS, TPF and SHG microscopy with a single 5 fs Ti:Sa laser