We show that fluorescence lifetime imaging microscopy (FLIM) of green fluorescent protein (GFP) molecules in cells can be used to report on the local refractive index of intracellular GFP. We expressed GFP fusion constructs of Rac2 and gp91(phox), which are both subunits of the phagocyte NADPH oxidase enzyme, in human myeloid PLB-985 cells and showed by high-resolution confocal fluorescence microscopy that GFP-Rac2 and GFP-gp91(phox) are targeted to the cytosol and to membranes, respectively. Frequency-domain FLIM experiments on these PLB-985 cells resulted in average fluorescence lifetimes of 2.70 ns for cytosolic GFP-Rac2 and 2.31 ns for membrane-bound GFP-gp91(phox). By comparing these lifetimes with a calibration curve obtained by measuring GFP lifetimes in PBS/glycerol mixtures of known refractive index, we found that the local refractive indices of cytosolic GFP-Rac2 and membrane-targeted GFP-gp91(phox) are approximately 1.38 and approximately 1.46, respectively, which is in good correspondence with reported values for the cytosol and plasma membrane measured by other techniques. The ability to measure the local refractive index of proteins in living cells by FLIM may be important in revealing intracellular spatial heterogeneities within organelles such as the plasma and phagosomal membrane.

The microwave dielectric properties of (BaPb)(ZrTi)O (BPZT) and ZnO thin films with thicknesses below were investigated. No significant dielectric relaxation was observed for both BPZT and ZnO up to 30 GHz. The intrinsic dielectric constant of BPZT was as high as 980 at 30 GHz. The absence of strong dielectric dispersion and loss peaks in the studied frequency range can be linked to the small grain diameters in these ultrathin films.

Lifetime and reliability in realistic operating conditions are important parameters for the application of thin-film piezoelectric microelectromechanical systems (piezoMEMS) based on lead zirconate titanate (PZT). Humidity can induce time-dependent dielectric breakdown at a higher rate compared to dry conditions, and significantly alter the dynamic behavior of piezoMEMS-devices. Here we assess the lifetime and reliability of PZT-based micromirrors with and without humidity barriers operated at 23 • C in an ambient of 0 and 95 % relative humidity. The correlation of the dynamic response, as well as the ferroelectric, dielectric, and leakage properties, with degradation time was investigated. In humid conditions, the median timeto-failure was increased from 2.7×10 4 [1.9×10 4 −4.0×10 4 ] s to 1.1×10 6 [0.9×10 6 −1.5×10 6 ] s at 20 V AC continuous unipolar actuation, by using a 40 nm thick Al 2 O 3 humidity barrier. However, the initial maximum angular deflection, polarization, and dielectric permittivity decreased by about 6, 11, and 12 %, respectively, for Al 2 O 3 capped devices. For both bare and encapsulated devices, the onset of electrothermal breakdown-events was the dominant cause of degradation. Severe distortions in the device's dynamic behavior, together with failure from loss of angular deflection, preceded time-dependent dielectric breakdown in 95% relative humidity. Moreover, due to the film-substrate stress transfer sensitivity of thin-film devices, water-induced degradation affects the reliability of thin-film piezoMEMS differently than bulk piezoMEMS. [2020-0228]

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