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Optoacoustic (OA) microscopy using an all-optical system based on the probe beam deflection technique (PBDT) for detection of laser-induced acoustic signals was investigated as an alternative to conventional piezoelectric transducers. PBDT provides a number of advantages for OA microscopy including (i) efficient coupling of laser excitation energy to the samples being imaged through the probing laser beam, (ii) undistorted coupling of acoustic waves to the detector without the need for separation of the optical and acoustic paths, (iii) high sensitivity and (iv) ultrawide bandwidth. Because of the unimpeded optical path in PBDT, diffraction-limited lateral resolution can be readily achieved. The sensitivity of the current PBDT sensor of 22 µV/Pa and its noise equivalent pressure (NEP) of 11.4 Pa are comparable with these parameters of the optical micro-ring resonator and commercial piezoelectric ultrasonic transducers. Benefits of the present prototype OA microscope were demonstrated by successfully resolving micron-size details in histological sections of cardiac muscle.

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The mechanism(s) responsible for the breakdown (nanoporation) of cell plasma membranes after nanosecond pulse (nsEP) exposure remains poorly understood. Current theories focus exclusively on the electrical field, citing electrostriction, water dipole alignment and/or electrodeformation as the primary mechanisms for pore formation. However, the delivery of a high-voltage nsEP to cells by tungsten electrodes creates a multitude of biophysical phenomena, including electrohydraulic cavitation, electrochemical interactions, thermoelastic expansion, and others. To date, very limited research has investigated non-electric phenomena occurring during nsEP exposures and their potential effect on cell nanoporation. Of primary interest is the production of acoustic shock waves during nsEP exposure, as it is known that acoustic shock waves can cause membrane poration (sonoporation). Based on these observations, our group characterized the acoustic pressure transients generated by nsEP and determined if such transients played any role in nanoporation. In this paper, we show that nsEP exposures, equivalent to those used in cellular studies, are capable of generating high-frequency (2.5 MHz), high-intensity (>13 kPa) pressure transients. Using confocal microscopy to measure cell uptake of YO-PRO®-1 (indicator of nanoporation of the plasma membrane) and changing the electrode geometry, we determined that acoustic waves alone are not responsible for poration of the membrane.

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In this work, the assembly of gold nanoparticles of (AuNPs) is used to detect the presence of the biomolecule glutathione (GSH) using a novel technique called "all-optical photoacoustic spectroscopy" (AOPAS). The AOPAS technique coupled with AuNPs forms the basis of a biosensing technique capable of probing the dynamic evolution of nano-bio interfaces within a microscopic volume. Dynamic Light Scattering (DLS) and ultraviolet-visible (UV-vis) spectra were measured to describe the kinetics governing the interparticle interactions by monitoring the AuNPs assembly and evolution of the surface plasmon resonance (SPR) band. A comparison of the same dynamic evolution of AuNPs assembly was performed using the AOPAS technique to confirm the validity of this method. The fundamental study is complemented by a demonstration of the performance of this biosensing technique in the presence of cell culture medium containing fetal bovine serum (FBS), which forms a protein corona on the surface of the AuNPs. This work demonstrates that the in vitro monitoring capabilities of the AOPAS provides sensitive measurement at the microscopic level and low nanoparticle concentrations without the artifacts limiting the use of conventional biosensing methods, such as fluorescent indicators. The AOPAS technique not only provides a facile approach for in vitro biosensing, but also shed a light on the real-time detection of thiol containing oxidative stress biomarkers in live systems using AuNPs.

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The goal of this paper is to demonstrate the unique capability of measuring the vector or angular information of propagating acoustic waves using an optical sensor. Acoustic waves were generated using photoacoustic interaction and detected by the probe beam deflection technique. Experiments and simulations were performed to study the interaction of acoustic emissions with an optical sensor in a coupling medium. The simulated results predict the probe beam and wavefront interaction and produced simulated signals that are verified by experiment.

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Optoacoustic imaging is an emerging medical technology that uniquely combines the absorption contrast of optical imaging and the penetration depth of ultrasound. While it is not currently employed as a clinical imaging modality, the results of current research strongly support the use of optoacoustic-based methods in medical imaging. One such application is the diagnosis of the presence of soft tissue foreign bodies. Because many radiolucent foreign bodies have sufficient contrast for imaging in the optical domain, laser-induced optoacoustic imaging could be advantageous for the detection of such objects. Common foreign bodies have been scanned over a range of visible and near infrared wavelengths by using an optoacoustic method to obtain the spectroscopic properties of the materials commonly associated with these foreign bodies. The derived optical absorption spectra compared quite closely to the absorption spectra generated when using a conventional spectrophotometer. By using the probe-beam deflection technique, a novel, pressure-wave detection method, we successfully generated optoacoustic spectroscopic plots of a wooden foreign body embedded in a tissue phantom, which closely resembled the spectrum of the same object obtained in isolation. A practical application of such spectra is to assemble a library of spectroscopic data for radiolucent materials, from which specific characteristic wavelengths can be selected for use in optimizing imaging instrumentation and provide a basis for the identification of the material properties of particular foreign bodies.

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Fluorescent nanodiamonds (FNDs) have drawn much attention in recent years for biomedical imaging applications due to their desired physical properties including excellent photostability, high biocompatibility, extended far-red fluorescence emission, and ease of surface functionalization. Here we explore a new feature of FNDs, i.e. their photoacoustic emission capability, which may lead to potential applications of using FNDs as a dual imaging contrast agent for combined fluorescence and photoacoustic imaging modalities. We observed significant enhancement of photoacoustic emission from FNDs when they were conjugated with gold nanoparticles (GNPs).

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BACKGROUND AND PURPOSE: Lower-pole ureteronephroscopy requires transmission of holmium:YAG energy along a deflected fiber. Current ureteroscopes are capable of high degrees of deflection, which may stress laser fibers beyond safe limits during lower-pole use. We hypothesized that optical fiber and safety measures differ among manufacturers. MATERIALS AND METHODS: Small (200-273-microm) and medium-diameter (300-400-microm) Ho:YAG fibers were tested in a straight and 180 degrees bent configuration. Energy transmission was measured by an energy detector. Fiber durability was assessed by firing the laser in sequentially tighter bending diameters. The fibers were bent to 180 degrees with a diameter of 6 cm and run at 200- to 4000-mJ pulse energy to determine the minimum energy required to fracture the fiber. The bending diameter was decreased by 1-cm increments and testing repeated until a bending diameter of 1 cm was reached. The maximum deflection of the ACMI DUR-8E ureteroscope with each fiber in the working channel was recorded. The flow rate through the working channel of the DUR-8E was measured for each fiber. RESULTS: The mean energy transmission differed among fibers (P < 0.001). The Lumenis SL 200 and the InnovaQuartz 400 were the best small and medium-diameter fibers, respectively, in resisting thermal breakdown (P < 0.01). The Dornier Lightguide Super 200 fractured repeatedly at a bend diameter of 2 cm and with the lowest energy (200 mJ). The other small fibers fractured only at a bend diameter of 1 cm. The Sharplan 200 and InnovaQuartz Sureflex 273T were the most flexible fibers, the Lumenis SL 365 the least. The flow rate was inversely proportional to four times the power of the diameter of the fiber. CONCLUSIONS: Optical performance and safety differ among fibers. Fibers transmit various amounts of energy to their cladding when bent. During lower-pole nephroscopy with the fiber deflected, there is a risk of fiber fracture from thermal breakdown and laser-energy transmission to the endoscope. Some available laser fibers carry a risk of ureteroscope damage.

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PURPOSE: The purpose of this study was to evaluate the toxicity and photodynamic activity of indocyanine green (ICG) and trypan blue (TryB) on cultured human lensepithelial cells (LECs). DESIGN: Experimental study. METHODS: Lens epithelial cell viability was assessed after treatment with ICG and TryB concentrations ranging from 0.025 to 5.0 mg/ml, and exposure to 806 nm diode laser. RESULTS: At ICG concentrations below 0.5 mg/ml, there was > or =75% cell viability; at higher ICG concentrations there was dose-dependent cytotoxicity in addition to loss of cellular viability due to ICG photosensitization. TryB had little cytotoxicity to the LECs: >80% cells were viable irrespective of the dye concentration or laser treatment. CONCLUSIONS: These data indicate that ICG may have application as a photosensitizer in the selective eradication of residual LECs after cataract surgery to reduce the incidence of posterior capsule opacification.

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Laser-induced heating in an ocular phantom is measured with magnetic resonance thermography (MRT) using temperature-dependent phase changes in proton resonance frequency. The ocular phantom contains a layer of melanosomes isolated from bovine retinal pigment epithelium. The phantom is heated by the 806-nm output of a continuous wave diode laser with an irradiance of 2.4 to 21.6 W/cm2 in a beam radius of 0.8 or 2.4 mm, depending on the experiment. MRT is performed with a 2 T magnet, and a two-turn, 6-cm-diam, circular radio frequency coil. Two-dimensional temperature gradients are measured within the plane of the melanin layer, as well as normal to it, with a temperature resolution of 1 degrees C or better. The temperature gradients extending within the melanin layer are broader than those orthogonal to the layer, consistent with the higher optical absorption and consequent heating in the melanin. The temperature gradients in the phantom measured by MRT closely approximate the predictions of a classical heat diffusion model. Three-dimensional temperature maps with a spatial resolution of 0.25 mm in all directions are also made. Although the temporal resolution is limited in the prototype system (22.9 s for a single image "slice"), improvements in future implementations are likely. These results indicate that MRT has sufficient spatial and temperature resolution to monitor target tissue temperature during transpupillary thermotherapy in the human eye.

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BACKGROUND AND OBJECTIVE: To demonstrate suitability of moiré interferometry to assess and quantify laser-induced shrinkage of scleral collagen for buckling procedures. MATERIALS AND METHODS: Scleral buckling of human cadaver eyes was investigated using a Coherent Ultrapulse CO2 laser. Projection moiré interferometry was employed to determine the out-of plane displacement produced by laser exposure, and in-situ optical microscopy of reference markers on the eye was used to measure in-plane shrinkage. RESULTS: Measurements based on moiré interferometry allow a three dimensional view of shape changes in the eye surface as laser treatment proceeds. Out-of-plane displacement reaches up to 1.5 mm with a single laser spot exposure. In-plane shrinkage reached a maximum of around 30%, which is similar to that reported by Sasoh et al (Ophthalmic Surg Lasers. 1998;29:410) for a Tm:YAG laser. CONCLUSION: The moiré technique is found to be suitable for quantifying the effects of CO2 laser scleral shrinkage and buckling. This can be further developed to provide a standardized method for experimental investigations of other laser sources for scleral shrinkage.

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