Polarization Z-Scan Studies Revealing Plasmon Coupling Enhancement Due to Dimer Formation of Gold Nanoparticles in Nematic Liquid Crystals.

We investigate the plasmon coupling of gold nanoparticle (AuNP) dimers dispersed in a nematic liquid crystal matrix using the polarization z-scan technique. Our experimental setup includes the precise control of incident light polarization through polarization angles of 0°, 45°, and 90°. Two distinct cell orientations are examined: parallel and twisted nematic cells. In parallel-oriented cells, where liquid crystal molecules and AuNPs align with the rubbing direction, we observe a remarkable 2-3-fold increase in the nonlinear absorption coefficient when the polarization of the incident light is parallel to the rubbing direction. Additionally, a linear decrease in the third-order nonlinear absorption coefficient is noted as the polarization angle varies from 0° to 90°. In the case of twisted nematic cells, the NPs do not have any preferred orientation, and the enhancement remains consistent across all polarization angles. These findings conclusively establish that the observed enhancement in the nonlinear absorption coefficient is a direct consequence of plasmon coupling, shedding light on the intricate interplay between plasmonic nanostructures and liquid crystal matrices.

Plasmon enhancement of third-order nonlinear optical absorption of gold nanoparticles dispersed in planar oriented nematic liquid crystals.

We report structural and nonlinear optical properties of 20 nm gold (Au) nanoparticles (NPs) that are dispersed in planar degenerate (non-oriented) and planar oriented nematic liquid crystals (LCs) (4'-Pentyl-4-biphenylcarbonitrile-5CB). Taking advantage of elastic forces in the planar oriented nematic LC, we aligned AuNPs parallel to the 5CB director axis. In the case of planar degenerate, 5CB is not aligned and has no preferred orientation, forcing the AuNPs to disperse randomly. Results show that the linear optical absorption coefficient for the planar oriented 5CB/AuNPs mixture is larger than the corresponding planar degenerate sample. The nonlinear absorption coefficients are greatly enhanced in planar oriented samples at relatively high concentrations which can be attributed to plasmon coupling between the aligned AuNPs. This study demonstrates the utility of LCs for developing the assembly of NPs with enhanced optical properties which may offer important insight and technological advancement for novel applications, including photonic nanomaterials and optoelectronic devices.

Electronic Characterization of Vis-NIR Absorbing Noninnocent Ruthenium Oxyquinolate-merocyanine Complexes and their Photoacoustic Emission in Aqueous Micellar Solution†.

The electronic and photophysical properties for a series of ruthenium(II) polypyridyl dyes are presented where a π-accepting 5-(vinyl-cyanine)-8-oxyquinolate class of ligand is incorporated to yield an improved vis-NIR absorption. A combination of computational, UV-vis-NIR absorption, phosphorescence emission and cyclic voltammetry studies are used to probe the influence of these ligands on complex electronic and photophysical properties. To assess their potential as vis-NIR photoacoustic contrast agents, select complexes were formulated in a PBS buffer/Tween® 20 solvent system. The p-quinolin-1-ium, 1,3,3-trimethyl-3H-indol-1-ium and 1,1,3-trimethyl-1H-benzo[e]indol-3-ium acceptor groups each impart a strong 680 nm optical absorption and photoacoustic emission on par with the performance exhibited by both the methylene blue and cryptocyanine commercial dyes.

Characterization of Triphenylamine and Ferrocenyl Donor-π-donor Vinyl BODIPY Derivatives as Photoacoustic Contrast Agents.

The photophysical and electrochemical properties for a series of BODIPY dyes with incremental 3- and 3,5-vinyl conjugation, as well as incremental electron-donating groups (anisole < triphenylamine < ferrocenyl), are presented. Insight into the influence of each vinyl-conjugated electron-donating group on both vis-NIR absorption and fluorescence emission properties is provided. These trends are further corroborated by density functional theory computational analysis. Two of this series containing the 3,5-bis(vinyltriphenylamine) and 3,5-bis(vinylferrocenyl) substituents exhibit significant absorption cross sections in the biological transparency window justifying further investigation of their photoacoustic emission properties via both optical photoacoustic z-scan and photoacoustic tomography experiments. Both the 3,5-bis(vinyltriphenylamine) and 3,5-bis(vinylferrocenyl) substituted BODIPY dyes exhibit quantitative photoacoustic quantum yields. Relative to the commercially available methylene blue and indocyanine green molecular photoacoustic contrast agents, the 3,5-bis(vinyltriphenylamine)-derived BODIPY exhibits the greatest photoacoustic emission and contrast upon excited-state absorption at 685 nm excitation at a low power laser fluence (<20 mJ cm-2 ).

Protease Degradation of Protein Coronas and Its Impact on Cancer Cells and Drug Payload Release.

The effect of matrix metalloproteinases (MMPs) on preformed protein coronas around spherical gold nanoparticles (AuNPs) was studied. Protein coronas of different compositions (human serum, human serum albumin, and collagen IV) were formed around AuNPs and characterized. The protease MMP-9 had different effects on the corona depending on the corona composition, resulting in different changes to the corona hydrodynamic diameter ( DH). When incubated with PANC-1 cells, the corona showed evidence of both increases as well as decreases in DH. Varying the composition of the corona influenced the MMP-9 activity. Furthermore, the corona was influenced not only by the protease activity of the MMP-9 but also by its ability to exchange with proteins in the preformed corona. This exchange could also occur with proteins in the media. Thus, the net effect of the MMP-9 was a combination of the MMP-9 protease activity and also exchange. Time scales for the exchange varied depending on the nature that make up the protein corona (weakly vs strongly bound corona proteins). Mass spectrometry was used to probe the protein corona composition and supported the exchange and degradation model. Together, these results indicate that the mechanism of protease activity on AuNP coronas involves both rearrangement and exchange, followed by degradation.

Photophysical and Photoacoustic Properties of π-Extended Curcumin Dyes. Effects of the Terminal Dimethylamino Electron-donor and the Bridging Aryl Ring.

The synthesis, photophysical and photoacoustic characterization for a series of nine π-extended quadrupolar curcumin dyes is presented. A systematic evaluation of the π-bridging unit including the p-phenyl, naphth-4-yl, thien-2-yl and hybrid 4-naphthathien-2-yl groups is presented. Furthermore, evaluation of the strongly donating donor-π-acceptor-π-donor quadrupolar dimethylamino terminated derivatives is also included. Select dyes exhibit excited state absorption at increased laser fluence which translates to the production of a nonlinear enhanced photoacoustic response. In particular, the bis-4-dimethylaminonaphtha-2-thien-5-yl curcuminBF2 contrast agent DMA-5 exhibits an excellent molar photoacoustics (PA) emission at both low (9.4 × 103  V M-1 ) and high (1.47 × 105  V M-1 ) laser fluence which is confirmed by its strong contrast by photoacoustic tomography (PAT). In summary, the strong absorbance and enhanced photoacoustic properties of naphthyl and thienyl curcuminoids here presented provides great promise for future photoacoustic imaging applications as demonstrated by preliminary PAT studies.

Characterization of a NIR absorbing thienyl curcumin contrast agent for photoacoustic imaging.

The synthesis and characterization of a bis(2-dimethylaminothien-5-yl)curcumin boron difluoride chromophore is presented. Photophysical, electrochemical and computational investigations establish the properties of its absorption in the Vis-NIR spectral range relative to established curcumin dyes. Application of this thienyl curcumin dye as a photoacoustic contrast agent is investigated against the dicarbocyanine Cy5 dye in the 675-735 nm excitation range.

Sex matters: females in proestrus show greater diazepam anxiolysis and brain-derived neurotrophin factor- and parvalbumin-positive neurons than males.

In humans and animal models, sex differences are reported for anxiety-like behavior and response to anxiogenic stimuli. In the current work, we studied anxiety-like behavior and response to the prototypical anti-anxiety drug, diazepam. We used 6th generation outbred lines of adult Long Evans rats with high and low anxiety-like behavior phenotypes to investigate the impact of proestrus on the baseline and diazepam-induced behavior. At three doses of diazepam (0, 0.1, and 1.0 mg/kg, i.p.), we measured anxiogenic responses on the elevated plus maze of adult male and female rats. We assessed parvalbumin and brain-derived neurotrophin protein levels in forebrain and limbic structures implicated in anxiety/stress using immunohistochemistry. At baseline, we saw significant differences between anxiety lines, with high anxiety lines displaying less time on the open arms of the elevated plus maze, and less open arm entries, regardless of sex. During proestrus, high anxiety females showed less anxiety-like behavior at 0.1 mg/kg, while low anxiety females displayed less anxiety-like behavior at 0.1 and 1.0 doses, relative to males. Brain-derived neurotrophin protein was elevated in females in the medial prefrontal cortex and central amygdala, while parvalbumin-immunoreactive cells were greater in males in the medial prefrontal cortex. Parvalbumin-positive cells in high anxiety females were higher in CA2 and dentate gyrus relative to males from the same line. In sum, when tested in proestrus, females showed greater anxiolytic effects of diazepam relative to males, and this correlated with increases in neurotrophin and parvalbumin neuron density in corticolimbic structures.

Photophysical and Photoacoustic Properties of Quadrupolar Borondifluoride Curcuminoid Dyes.

The synthesis and characterization of a series of donor-π-acceptor-π-donor (D-A-D) curcuminoid molecules is presented herein that incorporates π-extended aryl and electron-donating amino terminal functionalization. Computational evaluation shows these molecules possess quadrupolar character with the lowest energy transitions displaying high molar extinction coefficients with broad tunability through manipulation of terminal donating groups. Consistent with their quadrupolar nature, these molecules show varying degrees of solvatochromic behavior in both their absorption and emission spectra, which has been analyzed by Lippert-Mataga and Kamlet-Taft analysis. Photophysical and photoacoustic (PA) properties of these molecules have been investigated by the optical photoacoustic z-scan (OPAZ) method. Selected curcuminoid molecules display nonlinear behavior at a high laser fluence through excited state absorption that translates to the production of an enhanced photoacoustic emission. A relative comparison of "molar PA emission" is also presented with the crystal violet linear optical absorbing/linear PA emitting system being utilized as a standard reference material for OPAZ experiments. Furthermore, PA tomography experiments are presented to illustrate the enhanced PA contrast obtainable via an excited state absorption.

Digital holographic microscopy for longitudinal volumetric imaging of growth and treatment response in three-dimensional tumor models.

We report the use of digital holographic microscopy (DHM) as a viable microscopy approach for quantitative, nondestructive longitudinal imaging of in vitro three-dimensional (3-D) tumor models. Following established methods, we prepared 3-D cultures of pancreatic cancer cells in overlay geometry on extracellular matrix beds and obtained digital holograms at multiple time points throughout the duration of growth. The holograms were digitally processed and the unwrapped phase images were obtained to quantify the nodule thickness over time under normal growth and in cultures subject to chemotherapy treatment. In this manner, total nodule volumes are rapidly estimated and demonstrated here to show contrasting time-dependent changes during growth and in response to treatment. This work suggests the utility of DHM to quantify changes in 3-D structure over time and suggests the further development of this approach for time-lapse monitoring of 3-D morphological changes during growth and in response to treatment that would otherwise be impractical to visualize.

Shining light on the dark side of imaging: excited state absorption enhancement of a bis-styryl BODIPY photoacoustic contrast agent.

A first approach toward understanding the targeted design of molecular photoacoustic contrast agents (MPACs) is presented. Optical and photoacoustic Z-scan spectroscopy was used to identify how nonlinear (excited-state) absorption contributes to enhancing the photoacoustic emission of the curcuminBF2 and bis-styryl (MeOPh)2BODIPY dyes relative to Cy3.

Nonlinear optical properties of multipyrrole dyes.

The nonlinear optical properties of a series of pyrrolic compounds consisting of BODIPY and aza-BODIPY systems are investigated using 532 nm nanosecond laser and the Z-scan technique. Results show that 3,5-distyryl extension of BODIPY to the red shifted MeO2BODIPY dye has a dramatic impact on its nonlinear absorption properties changing it from a saturable absorber to an efficient reverse saturable absorbing material with a nonlinear absorption coefficient of 4.64 × 10-10 m/W. When plotted on a concentration scale per mole of dye in solution MeO2BODIPY far outperforms the recognized zinc(II) phthalocyanine dye and is comparable to that of zinc(II) tetraphenylporphyrin.

Digital holographic microscopy discriminates sex differences in medial prefrontal cortex GABA neurons following amphetamine sensitization.

Sex differences have been noted in patterns of drug use and relapse, and in particular with amphetamine abuse, implicating estradiol in mediating female neurobehavioral responses. To investigate the interaction of estradiol with amphetamine-induced hyperactivity, we compared male, intact female (INTACT), ovariectomized (OVX) and ovariectomized estradiol-treated (OVX+EB) female rats receiving repeated amphetamine (AMPH) treatment. All rats received intermittent AMPH injections for three days, and baseline and post-injection locomotor activity as well as fine-motor movements were recorded. Upon completion of behavioral experiments, immunohistochemistry was performed to assess parvalbumin-immunoreactive (PV-IR) GABAergic neurons in the medial prefrontal cortex (mPFC). Results indicate that AMPH induced greater behavioral response during habituation among the INTACT animals, and post-injection hyperactivity was apparent on days 2 and 3, among INTACT and OVX+EB females. For INTACT animals, the hyperactivity was most pronounced when estrogen levels were high. Immunohistochemical analysis using digital holographic microscopy revealed INTACT and OVX+EB females had less expression and smaller somatic area of PV-IR neurons in the mPFC. These data provide evidence for rapid development of sex differences in response to AMPH that correlates with sexually dimorphic alterations in a subset of mPFC GABAergic neurons implicated in modulating forebrain dopamine projections.

Parallel-quadrature phase-shifting digital holographic microscopy using polarization beam splitter.

We present a digital holography microscopy technique based on parallel-quadrature phase-shifting method. Two π/2 phase-shifted holograms are recorded simultaneously using polarization phase-shifting principle, slightly off-axis recording geometry, and two identical CCD sensors. The parallel phase-shifting is realized by combining circularly polarized object beam with a 45° degree polarized reference beam through a polarizing beam splitter. DC term is eliminated by subtracting the two holograms from each other and the object information is reconstructed after selecting the frequency spectrum of the real image. Both amplitude and phase object reconstruction results are presented. Simultaneous recording eliminates phase errors caused by mechanical vibrations and air turbulences. The slightly off-axis recording geometry with phase-shifting allows a much larger dimension of the spatial filter for reconstruction of the object information. This leads to better reconstruction capability than traditional off-axis holography.

Quantitative phase microscopy using dual-plane in-line digital holography.

We present detailed theoretical evaluation and thorough experimental investigation of quantitative phase imaging using our previously demonstrated dual-plane in-line digital holographic microscopy technique [Opt. Lett. 35, 3426 (2010)]. This evaluation is based on the recording of two interferograms at slightly different planes and numerically reconstructing the object information. The zero-order diffracted wave is eliminated by using the method of subtraction of average intensity of the entire hologram, and the twin-image diffracted wave is removed by Fourier domain processing of the two recorded holograms. Experiments are performed using controlled amplitude and phase objects and human muscle cells to demonstrate the potential of this technique.

Dual-channel in-line digital holographic double random phase encryption.

We present a robust encryption method for the encoding of 2D/3D objects using digital holography and virtual optics. Using our recently developed dual-plane in-line digital holography technique, two in-line digital holograms are recorded at two different planes and are encrypted using two different double random phase encryption configurations, independently. The process of using two mutually exclusive encryption channels makes the system more robust against attacks since both the channels should be decrypted accurately in order to get a recognizable reconstruction. Results show that the reconstructed object is unrecognizable even when the portion of the correct phase keys used during decryption is close to 75%. The system is verified against blind decryptions by evaluating the SNR and MSE. Validation of the proposed method and sensitivities of the associated parameters are quantitatively analyzed and illustrated.

Dual plane in-line digital holographic microscopy.

We report a dual plane in-line digital holographic microscopy technique that exploits the method of subtraction of average intensity of the entire hologram to suppress the zero-order diffracted wave. Two interferograms are recorded at different planes to eliminate the conjugate image. The experimental results demonstrate successful reconstruction of phase objects as well as of amplitude objects. The two interferograms can be recorded simultaneously, using two CCD or CMOS sensors, in order to increase the acquisition rate. This enhanced acquisition rate, together with the improved reconstruction capability of the proposed method, may find applications in biomedical research for visualization of rapid dynamic processes at the cellular level.

Nonlinear photoacoustics for measuring the nonlinear optical absorption coefficient.

We report a novel photoacoustic Z-scan (PAZ-scan) technique that combines the advantages offered by the conventional Z-scan method and the sensitivity of the photoacoustic detection. The sample is scanned through the focused laser beam and the generated photoacoustic signal is recorded using a 10 MHz focused ultrasound transducer. Since the signal strength is directly proportional to the optical absorption, PAZ-scan displays nonlinear behavior depicting the nonlinear optical absorption of the material. Among many advantages, our experiments on mouse blood show that PAZ-scan can potentially be used as a standard technique to calibrate contrast agents used in theranostics in general and photoacoustics in particular.

Common-path multimodal optical microscopy.

We have developed a common-path multimodal optical microscopy system that is capable of using a single optical source and a single camera to image amplitude, phase, and fluorescence features of a biological specimen. This is achieved by varying either contrast enhancement filters at the Fourier plane and/or neutral density/fluorescence filters in front of the CCD camera. The feasibility of the technique is demonstrated by obtaining brightfield, fluorescence, phase-contrast, spatially filtered, brightfield+fluorescence, phase+fluorescence, and edge-enhanced+fluorescence images of the same Drosophila embryo without the need for image registration and fusion. This comprehensive microscope has the capability of providing both structural and functional information and may be used for applications such as studying live-cell dynamics and in high throughput microscopy and automated microscopy.

Coherent population oscillations and superluminal light in a protein complex.

We observed superluminal light in aqueous solution of the protein complex bacteriorhodopsin (bR) at 647.1 nm wavelength where it exhibits reverse saturable behavior, exploiting the technique of coherent population oscillations (CPO). With a modulation frequency of 10 Hz, the signal pulse through a 1 cm path cell is ahead by 3 msec relative to the reference pulse, corresponding to a group velocity of -3.3 m/sec. Following our early work on slow light in the same sample at the saturable wavelength 568.2 nm, we now explicitly observed the narrow spectral hole in the absorption band of the stable B state and further, demonstrated a close correlation between the profile of the hole and the corresponding pulse delay for various modulation frequencies. A similar behavior is observed for superluminal light versus antihole blown in the absorption band.