This PDF file contains the front matter associated with SPIE Proceedings Volume 9321, including the Title Page, Copyright information, Table of Contents, Introduction (if any), and Conference Committee listing.

Laser-induced formation of pores in cartilage and sclera tissues is a basis of novel technologies for treatment of arthritis and glaucoma. The presented theoretical model describes dynamics of the pore formation and allows optimizing laser settings.

Collagen cross-linking in cornea has the capability of enhancing its mechanical properties and thereby providing an alternative treatment for eye diseases such as keratoconus. Currently, riboflavin assisted UVA light irradiation is a method of choice for cross-link induction in eyes. However, ultrafast pulsed laser interactions may be a powerful alternative enabling in-depth treatment while simultaneously diminishing harmful side effects such as, keratocyte apoptosis. In this study, femtosecond laser is utilized for treatment of bovine cornea slices. It is hypothesized that nonlinear absorption of femtosecond laser pulses plays a major role in the maturation of immature cross-links and the promotion of their growth. Targeted irradiation with tightly focused laser pulses allows for the absence of a photosensitizing agent. Inflation test was conducted on half treated porcine cornea to identify the changes of mechanical properties due to laser treatment. Raman spectroscopy was utilized to study subtle changes in the chemical composition of treated cornea. The effects of treatment are analyzed by observing shifts in Amide I and Amide III bands, which suggest deformation of the collagen structure in cornea due to presence of newly formed cross-links.

Attachment of single cells via hemifusion of cellular membranes using femtosecond laser pulses is reported in this manuscript. This is a method to attach single cells using sub-10 femtosecond laser pulses, with 800 nm central wavelength delivered from a Ti:Sapphire laser is described. A fluorescent dye, Calcein AM, was used to verify that the cell’s cytoplasm did not migrate from a dyed cell to a non-dyed cell, in order to ascertain that the cells did not go through cell-fusion process. An optical tweezer was used in order to assess the mechanical integrity of the attached joint membranes. Hemifusion of cellular membranes was successful without initiating full cell fusion. Attachment efficiency of 95% was achieved, while the cells’ viability was preserved. The attachment was performed via the delivery of one to two trains of sub-10 femtosecond laser pulses lasting 15 milliseconds each. An ultrafast reversible destabilization of the phospholipid molecules in the cellular membranes was induced due to a laser-induced ionization process. The inner phospholipid cell membrane remained intact during the attachment procedure, and cells’ cytoplasm remained isolated from the surrounding medium. The unbounded inner phospholipid molecules bonded to the nearest free phospholipid molecule, forming a joint cellular membrane at the connection point. The cellular membrane hemifusion technique can potentially provide a platform for the creation of engineered tissue and cell cultures.

Laser assisted corneal surgeries often rely on the nonlinear absorption effect of ultrafast lasers to induce features in the interior of the cornea without affecting the surface. In particular, corneal flap formation in femtosecond assisted Laser- Assisted in situ Keratomileusis (LASIK) is based on the bubble creation. This study focuses on the interaction between the tissue and the femtosecond laser. Interior of cornea is treated with tightly focused femtosecond laser pulses. Due to the nature of the process, heating of the tissue within and around the focal volume is practically instantaneous. The affected region is subject to thermoelastic stress that arises with the steep temperature elevation. To predict the size of the region subject to the morphological changes due to the laser treatment, the temperature field is calculated. Cavitation bubble initiation and expansion process, which acts as precursor to the stress induced tissue trauma, is studied as well. Theoretical findings are compared against experimental results. High-speed camera is utilized to assess the laser treatment process, showing the temporal development of the cavitation bubbles. The results obtained in this study facilitate a better understanding of the effects of femtosecond laser assisted corneal surgeries and help in choosing optimal laser parameters.

Transparent ocular tissues such as cornea and crystalline lens can be precisely ablated or dissected using ultrafast ultraviolet, visible, and infrared lasers. In refractive or cataract surgery, cutting of the cornea, lens, and lens capsule is typically produced by dielectric breakdown in the focus of a short-pulse laser which results in explosive vaporization of the interstitial water and mechanically ruptures the surrounding tissue. Here, we report that tissue can also be disrupted below the threshold of bubble appearance using 400 nm femtosecond pulses with minimal mechanical damage. Using gel electrophoresis and liquid chromatography/mass spectrometry, we assessed photodissociation of proteins and polypeptides by 400 nm femtosecond pulses both below and above the cavitation bubble threshold. Negligible protein dissociation was observed with 800 nm femtosecond lasers even above the threshold of dielectric breakdown. Scanning electron microscopy of the cut edges in porcine lens capsule demonstrated that plasma-mediated cutting results in the formation of grooves. Below the cavitation bubble threshold, precise cutting could still be produced with 400 nm femtosecond pulses, possibly due to molecular photodissociation of the tissue structural proteins.

Thresholds for microcavitation of isolated bovine retinal melanosomes were determined as a function of temperature using single nanosecond laser pulses at 532 nm and 1064 nm. Melanosomes were irradiated using a 1064-nm Qswitched Nd:YAG (doubled for 532-nm irradiation). Time-resolved microscopy was accomplished by varying the delay between the irradiation beam and an illumination beam allowing stroboscopic imaging of microcavitation events. Results indicated a decrease in microcavitation fluence threshold with increasing sample temperature for both 532-nm and 1064-nm single pulse exposures. The nucleation temperature at both wavelengths was extrapolated through the linear relationship between the temperature increases and the decrease in fluence threshold. In addition, absorption coefficients of melanosomes for visible and near-infrared wavelengths were estimated using the calculated nucleation temperatures.

In laser tissue soldering (LTS) protein solutions are used for closing of incisions or fixation of wound dressings. During coagulation and thermal denaturation of the protein solutions their morphology changes significantly such that light is strongly scattered. When scattering becomes major component extinction increases and the optical penetration depth shrinks which could lead to unsufficient coagulation and bonding. For adaption of extinction during coagulation we are investigating a two-wavelength approach. A strongly absorbed laser wavelength (1540 nm) and weakly absorbed wavelength (980 nm) can be applied simultaneously. Simulation of beam propagation is performed in natural and coagulated state of the solder. The model describes a three-layer system consisting of membrane, solder and phantom. The optical properties are determined by spectrometric measurements both in natural and coagulated state. The absorption coefficient μ_a, scattering coefficient μ_s and anisotropy factor γ are determined by numerical analysis from the spectrometric data. Beam propagation is simulated for 980 nm and 1540 nm radiation with ZEMAX^® software based on the Monte Carlo method. For both wavelengths the beginning of the process with a clear solder layer, and the final state characterized by a coagulated solder layer are examined. The optical penetration depth depends mainly on the optical properties of the solder, which change in the course of coagulation process. The coagulation depth can be varied between 1.5 mm to 3.5 mm by changing the proportion of both laser sources. This leads to concepts for minimizing heat input while maintaining a constant coagulation depth.

Multiple femtosecond lasers have now been cleared for use for ophthalmic surgery, including for creation of corneal flaps in LASIK surgery. Preliminary study indicated that during typical surgical use, laser energy may pass beyond the cornea with potential effects on the iris. As a model for laser exposure of the iris during femtosecond corneal surgery, we simulated the temperature rise in porcine cadaver iris during direct illumination by the femtosecond laser. Additionally, ex-vivo iris heating due to femtosecond laser irradiation was measured with an infrared thermal camera (Fluke corp. Everett, WA) as a validation of the simulation.

Atherosclerotic plaques mainly consist of cholesteryl esters. Cholesteryl esters have an absorption peak at the wavelength of 5.75 μm originated from C=O stretching vibration mode of ester bond. Our group achieved making cutting difference between atherosclerotic lesions and normal vessels using a quantum cascade laser (QCL) in the 5.7 μm wavelength range. QCLs are relatively new types of semiconductor lasers that can emit mid-infrared range. They are sufficiently compact and have recently achieved their high-power emission. However, large thermal damage was observed because the QCL worked as a quasi-continuous wave laser due to its short pulse interval. To realize less invasive ablation by the QCL, reducing thermal effects to normal vessels is needed. In this study, we tried improving the thermal effects by changing the pulse structure. First, irradiation effects to rabbit atherosclerotic aortas by macro pulse irradiation (irradiation of pulses at intervals) and conventional continuous pulse irradiation were compared. The macro pulse width and the macro pulse interval were set to 0.54 and 12 ms, respectively, because the thermal relaxation time of rabbit normal and atherosclerotic aortas in the oscillation wavelength was 0.54-12 ms. As a result, ablation depth became longer and coagulation width became shorter by the macro pulse irradiation. In addition, cutting difference between rabbit normal and atherosclerotic aortas was observed by the macro pulse irradiation. Therefore, the macro pulse irradiation achieved the improvement of thermal effects by the QCL in the 5.7 μm wavelength range. The QCL has the potential of realizing less-invasive laser angioplasty.

The viscoelasticity of skin is an important indicator of its healthy condition. However, monitoring the mechanical properties is usually invasive and destructive. In this report, we employed Brillouin microspectroscopy to assess changes of viscoelastic properties of various skin samples. To induce skin injuries, we used the high-power laser irradiation. Brillouin spectra were collected by an earlier developed background free virtually imaged phased array (VIPA) spectrometer. Raman spectroscopy measurements were used to supplement local viscoelastic assessment of skin injuries.

The time-temperature effects of laser radiation exposure are investigated as a function of wavelength. We experimentally measure the thermal response of tissue to laser radiation ranging in wavelength from 1100 nm to 1550 nm. Simulations were then performed to estimate damage thresholds.

Laser damage thresholds were determined for exposure to 2.5-ms 532-nm pulses in an established in vitro retinal model. Single and multiple pulses (10, 100, 1000) were delivered to the cultured cells at three different pulse repetition frequency (PRF) values, and overt damage (membrane breach) was scored 1 hr post laser exposure. Trends in the damage data within and across the PRF range identified significant thermal additivity as PRF was increased, as evidenced by drastically reduced threshold values (< 40% of single-pulse value). Microthermography data that were collected in real time during each exposure also provided evidence of thermal additivity between successive laser pulses. Using thermal profiles simulated at high temporal resolution, damage threshold values were predicted by an in-house computational model. Our simulated ED₅₀ value for a single 2.5-ms pulse was in very good agreement with experimental results, but ED₅₀ predictions for multiple-pulse trains will require more refinement.

Melanin particles often present as an aggregate of smaller melanin pigment granules and have a heterogeneous surface morphology. When irradiated with light within the absorption spectrum of melanin, these heterogeneities produce measurable concentrations of the electric field that result in temperature gradients from thermal effects that are not seen with spherical or ellipsoidal modeling of melanin. Modeling melanin without taking into consideration the heterogeneous surface morphology yields results that underestimate the strongest signals or over{estimate their spatial extent. We present a new technique to image phase changes induced by heating using a computational model of melanin that exhibits these surface heterogeneities. From this analysis, we demonstrate the heterogeneous energy absorption and resulting heating that occurs at the surface of the melanin granule that is consistent with three{photon absorption. Using the three{photon dluorescence as a beacon, we propose a method for detecting the extents of the melanin granule using photothermal microscopy to measure the phase changes resulting from the heating of the melanin.

We studied the myocardial cell necrosis measuring cell lethality with various albumin concentrations against oxidative stress by extracellular photosensitization reaction using talaporfin sodium. We have proposed to induce the photosensitization reaction in interstitial space of myocardium with short drug-light interval to realize the immediate cell damage by myocardial cell membrane for tachyarrhythmia treatment. To understand the effect of the particular extracellular photosensitization reaction in interstitial space, we measured the myocardial cell lethality 2 hours after the photosensitization reaction by WST assay varying the talaporfin sodium concentration, radiant exposure, and wide albumin concentration of 0-15 mg/ml. The cell lethality was decreased with albumin concentration increasing over 85 % in binding ratio between talaporfin sodium and albumin. The cell lethality didn’t change between 0-85% in binding ratio between talaporfin sodium and albumin. The calculated deposited energy to the talaporfin sodium solution was nearly constant of 8.4±1.6 J/well in average varying albumin concentration. We think the cell killing effect by the extracellular photosensitization reaction has a threshold between 85-100% in the binding ratio between talaporfin sodium and albumin in the case of the talaporfin sodium concentration of 40 μg/ml and radiant exposure of 0-40 J/cm².

Low intensity laser irradiation of photoactive ligands bound non-covalently to proteins can generate a structural change in the proteins, which is detectable spectroscopically. This light induced protein modification could help to study the structure/function relationship in proteins or to prompt non-native protein properties. That is, only if we can determine if and how protein function is effected. Much work has shown small light-induced secondary and tertiary structural changes to albumin have occurred when the protein is bound to a porphyrin such as protoporphyrin IX or meso-tetra(4- sulfonatophenyl)porphyrin (TSPP) and irradiated. This Affinity-Depletion study aims to explore the conformational change of TSPP-bound albumin after visible-light irradiation by testing its ability to bind the biologically relevant albumin receptor, osteonectin. Osteonectin has been covalently attached to magnetic beads, forming an affinity column, but after ten trials (of varied protocol) no substantial albumin-to-osteonectin binding could be achieved.

Cellular delivery of pulsed IR laser energy has been shown to stimulate action potentials in neurons. The mechanism for this stimulation is not completely understood. Certain hypotheses suggest the rise in temperature from IR exposure could activate temperature- or pressure-sensitive channels, or create pores in the cellular outer membrane. Studies using intensity-based Ca^2+-responsive dyes show changes in Ca²⁺ levels after various IR stimulation parameters; however, determination of the origin of this signal proved difficult. An influx of larger, typically plasma-membrane-impermeant ions has been demonstrated, which suggests that Ca²⁺ may originate from the external solution. However, activation of intracellular signaling pathways, possibly indicating a more complex role of increasing Ca²⁺ concentration, has also been shown. By usingCa²⁺ sensitive dye Fura-2 and a high-speed ratiometric imaging system that rapidly alternates the excitation wavelengths, we have quantified the Ca²⁺ mobilization in terms of influx from the external solution and efflux from intracellular organelles. CHO-K1 cells, which lack voltage-gated Ca²⁺ channels, and NG-108 neuroblastoma cells, which do not produce action potentials in an early undifferentiated state, are used to determine the origin of the Ca²⁺ signals and investigate the role these mechanisms may play in IR neural stimulation.

We studied time response of electrical conduction (EC) block in a novel cardiomyocyte wire by extra-cellular photosensitization reaction (EPR) at various irradiances. This EC block using the EPR has been studied to develop a non-thermal arrhythmia therapeutic methodology. Despite the EC block in acute phase is needed to judge therapeutic endpoint in clinical arrhythmia therapy, time response of the EC block by the EPR in acute phase hasn’t been studied. We measured the time to EC block occurrence by the EPR with intra-cellular Ca²⁺ concentration change using Fluo-4 AM fluorescence measurement by a confocal laser microscope system. The pattern cultivation cover glass with 10 mm Φ which had 60 μm width cultivation areas with 300 μm separations was used to form the cardiomyocyte wires. Rat cardiomyocyte with 10.8×105 cells was disseminated to the cover glasses installed in a 35 mmΦ dish. After 3 days from the dissemination, the EPR was operated to the cardiomyocyte wires for 10 min varying 3-120 mW/cm² in 663 nm laser irradiances with 20 μg/ml talaporfin sodium. An irradiation area was approximately 60×340 μm² on each wires. Cross correlation functions (CCF) in measured fluorescence images in every 10 s were calculated across the irradiation area. The time to EC block occurrence was defined as the time of the max difference between adjacent CCFs. By decreasing irradiances in 30-6 mW/cm², the time to EC block occurrence became longer from 294 to 434 s. In 30-120 mW/cm², the time to EC block occurrence was nearly constant in 300 s.

Terahertz (THz) spectroscopy and imaging of biomedical samples is expected to be an important application of THz analysis techniques. Identification and localization of tumor tissue, imaging of biological samples, and analysis of DNA by THz spectroscopy have been reported. THz time-domain spectroscopy (TDS) is useful for obtaining the refractive index over a broad frequency range. However, THz-TDS spectra of fresh tissue samples are sensitive to procedures such as sample preparation, and a standardized measurement protocol is required. Therefore, in this work, we establish a protocol for measurements of THz spectra of fresh tissue and demonstrate reliable detection of rat brain tumor tissue. We use a reflection THz-TDS system to measure the refractive index spectra of the samples mounted on a quartz plate. The tissue samples were measured immediately after sectioning to avoid sample denaturalization during storage. Special care was taken in THz data processing to eliminate parasitic reflections and reduce noise. The error level in our refractive index measurements was as low as 0.02 in the frequency range 0.8–1.5 THz. With increasing frequency, the refractive index in the tumor and normal regions monotonically decreased, similarly to water, and it was 0.02 higher in the tumor regions. The spectral data suggest that the tumor regions have higher water content. Hematoxylin-eosin stained images showed that increased cell density was also responsible for the observed spectral features. A set of samples from 10 rats showed consistent results. Our results suggest that reliable tumor detection in fresh tissue without pretreatment is possible with THz spectroscopy measurements. THz spectroscopy has the potential to become a real-time in vivo diagnostic method.

In recent years, a surge in the development of many terahertz (THz) sensing and imaging technologies occurred leading to increased use in military and civil operations. Therefore, understanding the biological effects associated with exposures to this radiation is becoming increasingly important. Previous studies have speculated that cells exposed to different frequencies of THz radiation may exhibit differential responses. However, empirical studies to confirm such differences have not been performed. The question of whether cells exposed to different THz frequencies exhibited specific biological responses remains unclear. In this study, we exposed human keratinocytes to a THz laser tuned to several different THz frequencies using our recently developed THz exposure system. This system consists of an optically pumped molecular gas THz laser source coupled to a modified cell culture incubator permitting THz radiation exposures under controlled standard tissue culture conditions. For all frequencies, we matched the THz exposure duration and irradiance. During THz exposure, we monitored the power as DC voltage-logged values (LabVIEW™ IV log). To determine the temperature changes by THz exposure, we collected temperature readings from the unexposed and THz-exposed cells using thermocouples. We assessed cellular viability after exposure using MTT colorimetric assays. We compared the changes in gene expression profiles using messenger RNA (mRNA) microarrays, and we identified the THz-induced signaling pathways for each frequency using bioinformatics. Our data provide valuable new insights that give a comparative picture of the genes and intracellular signaling pathways triggered in cells exposed to THz radiation at different frequencies.

Optical clearing allows the reduction of light scattering in biological tissue, enabling 3D morphological information to be obtained deep within tissue using techniques such as optical projection tomography and light sheet microscopy. However, the extent of the clearing is dependent on the technique that is used. There is therefore a need for methods to quantify the quality of the clearing process and thereby to compare clearing techniques. In this study, we developed such a method using a custom spectroscopy system and applied it to compare three techniques that were applied to mouse brain: BABB (Murray’s clear), pBABB (a modification of BABB which includes the use of hydrogen peroxide), and passive CLARITY.

We are studying the transmission of LED array-emitted near-infrared (NIR) light through human tissues. Herein, we simulated and measured transcranial NIR penetration in highly scattering human head tissues. Using finite element analysis, we simulated photon diffusion in a multilayered 3D human head model that consists of scalp, skull, cerebral spinal fluid, gray matter and white matter. The optical properties of each layer, namely scattering and absorption coefficient, correspond to the 850 nm NIR light. The geometry of the model is minimally modified from the IEEE standard and the multiple LED emitters in an array were evenly distributed on the scalp. Our results show that photon distribution produced by the array exhibits little variation at similar brain depth, suggesting that due to strong scattering effects of the tissues, discrete spatial arrangements of LED emitters in an array has the potential to create a quasi-radially symmetrical illumination field. Measurements on cadaveric human head tissues excised from occipital, parietal, frontal and temporal regions show that illumination with an 850 nm LED emitter rendered a photon flux that closely follows simulation results. In addition, prolonged illumination of LED emitted NIR showed minimal thermal effects on the brain.

Under the assumption of high scattering and weak absorbing media, diffusion approximation holds in the radiative transport equation to model propagation of light. Diffusion approximation is valid deep inside the medium, not near the boundary. So, we need to implement accurate boundary conditions. Diffuse reflectance close to the source, majorly, depends on the source model inside the medium and boundary conditions used to derive the analytical solution. We have implemented partial current boundary condition and extrapolated boundary condition with extended isotropic source (exponentially decaying) model. Our model predicts diffuse reflectance close to the source at distance less than one mean free path is more accurate than the other methods. Monte-carlo simulation is the standard model to provide diffuse reflectance close to source most accurately. In this report, partial current, extrapolated boundary condition and a unified boundary condition have been compared for accuracy at different regions from the source. It is found that different boundary conditions work in different regimes and the relative error is less with extended source compared to point source.

Optical contrast agents introduce distinct features to induce detectable changes in native tissue properties [1]. In ultrasound imaging, microbubbles (MBs) – a gas-core shell-encapsulated agent – are used clinically as contrast agents. The working hypothesis of this study is that microbubbles can be employed as an intravascular contrast agent in optical imaging systems. Microbubbles can produce a refractive index mismatch which makes it distinguishable from surrounding media. In this work, the interaction of collimated light and microbubbles in a [1] biological phantom solution was investigated. The biological medium was comprised of intralipid and human blood which was constructed to cover the range of soft tissue optical properties. The effect of microbubbles on the optical properties such as reduced scattering and absorption coefficients were considered. Diffuse reflectance (DR) and total transmittance (TT) of a biological phantom solution were measured using a spectroscopic integrating sphere system in the absence and presence of Definity® microbubbles. The optical properties were computed using the inverse adding doubling (IAD) software. The presence of microbubbles increased DR and decreased TT of the phantom. In the presence of MB’s (2.5% volume concentration), the reflectance of the phantom increased by 25% in the optical window. There is no absorption event and only scattering happened after light-microbubbles interactions. The reduced scattering coefficient increased significantly (30%) indicating the potential use of MBs as optical contrast agents. In conclusion, reflectance of a media can be enhanced by adding microbubbles to increase scattering properties and more light was detected returning to the surface of tissue.

Clinical protocols are recommended in device guidelines outlined for treating many diseases on empirical basis. However, effects of low-intensity infrared lasers at fluences used in clinical protocols on DNA are controversial. Excitation of endogenous chromophores in tissues and free radicals generation could be described as a consequence of laser used. DNA lesions induced by free radicals cause changes in DNA structure, chromatin organization, ploidy degrees and cell death. In this work, we investigated whether low-intensity infrared laser therapy could alter the fibroblasts nuclei characteristics and induce DNA fragmentation. Tendons of Wistar rats were exposed to low-intensity infrared laser (830 nm), at different fluences (1, 5 and 10 J/cm²), in continuous wave (power output of 10mW, power density of 79.6 mW/cm²). Different frequencies were analyzed for the higher fluence (10 J/cm²), at pulsed emission mode (2.5, 250 and 2500 Hz), with the laser source at surface of skin. Geometric, densitometric and textural parameters obtained for Feulgen-stained nuclei by image analysis were used to define nuclear phenotypes. Significant differences were observed on the nuclear phenotype of tendons after exposure to laser, as well as, high cell death percentages was observed for all fluences and frequencies analyzed here, exception 1 J/cm² fluence. Our results indicate that low-intensity infrared laser can alter geometric, densitometric and textural parameters in tendon fibroblasts nuclei. Laser can also induce DNA fragmentation, chromatin lost and consequently cell death, using fluences, frequencies and emission modes took out from clinical protocols.

Application of light spectroscopy based techniques for the detection of cancers have emerged as a promising approach for tumor diagnostics. In-vivo or freshly excised samples are normally used for point spectroscopic studies. However, ethical issues related to in-vivo studies, rapid decay of surgically excised tissues and sample availability puts a limitation on in-vivo and in-vitro studies. There has been a few studies reported on the application of formalin fixed samples with good discrimination capability. Usually formalin fixation is performed to prevent degradation of tissues after surgical resection. Fixing tissues in formalin prevents cell death by forming cross-linkages with proteins. Previous investigations have revealed that washing tissues fixed in formalin using phosphate buffered saline is known to reduce the effects of formalin during spectroscopic measurements. But this could not be the case with reflectance measurements. Hemoglobin is a principal absorbing medium in biological tissues in the visible range. Formalin fixation causes hemoglobin to seep out from red blood cells. Also, there could be alterations in the refractive index of tissues when fixed in formalin. In this study, we propose to investigate the changes in tissue optical properties between freshly excised and formalin fixed brain tissues. The results indicate a complete change in the spectral profile in the visible range where hemoglobin has its maximum absorption peaks. The characteristic bands of oxy-hemoglobin at 540, 580 nm and deoxy-hemoglobin at 555 nm disappear in the case of samples fixed in formalin. In addition, an increased spectral intensity was observed for the wavelengths greater than 650 nm where scattering phenomena are presumed to dominate.

The goal of this study was to evaluate the dynamic process of water-mediated hard dental tissue ablation induced by Ho:YAG laser with high-speed camera. Human molars in vitro of yellow race were cut into tooth sections and irradiated with pulsed Ho:YAG laser with a wavelength of 2.08μm. The pulse repetition rate was 3 Hz and laser energy ranged from 300 to 2000 mJ. The frame rate of high-speed camera used in the experiment was 50525 fps. Based on the observation by high-speed camera, the dynamic process of the oscillating cavitation bubble and water-mediated ablation induced by Ho:YAG laser was efficiently recorded and graphically described. The pulsation period, the maximum length and width of vapor channel increased with laser energy. The results showed that the external water played multiple roles in laser ablation of hard dental tissue, not only acting as a channel to transmit laser energy, but also helping to improve the regularity of the ablation shape.

The aim of this in vitro study was to evaluate the influence of low-level laser irradiation (LLLI) on the chondrocytes proliferation cultured in different concentration of fetal bovine serum (FBS) using 658 nm, 785 nm and 830 nm diode lasers. The role of energy density (10-70 mJ·cm^-2) on chondrocytes proliferation following irradiation with 658 nm laser for 2 days was firstly investigated to find out the best laser energy density. Then the effect of LLLI on the proliferation of chondrocytes cultured with fetal bovine serum at 0%, 2%, 5% and 10% was also evaluated. The results showed that there was no or little photobiostimulation on the proliferation of chondrocytes cultured with 0% FBS and 10% FBS; the cell proliferation at 2% and 5% FBS was significantly modulated by LLLI.

Retinal laser photocoagulation is an established and successful treatment for a variety of retinal diseases. While being a valuable treatment modality, laser photocoagulation shows the drawback of employing high energy lasers which are capable of physically destroying the neural retina. For reliable therapy, it is therefore crucial to closely monitor the therapy effects caused in the retinal tissue. A depth resolved representation of optical tissue properties as provided by optical coherence tomography may provide valuable information about the treatment effects in the retinal layers if recorded simultaneously to laser coagulation. Therefore, in this work, the use of ultra-high resolution optical coherence tomography to represent tissue changes caused by conventional and selective retinal photocoagulation is investigated. Laser lesions were placed on porcine retina ex-vivo using a 577 nm laser as well as a pulsed laser at 527 nm built for selective treatment of the retinal pigment epithelium. Applied energies were varied to generate lesions best representing the span from under- to overtreatment. The lesions were examined using a custom-designed optical coherence tomography system with an axial resolution of 1.78 μm and 70 kHz Ascan rate. Optical coherence tomography scans included volume scans before and after irradiation, as well as time lapse scans (Mscan) of the lesions. Results show OCT lesion visibility thresholds to be below the thresholds of ophthalmoscopic inspection. With the ultra-high resolution OCT, 42% - 44% of ophthalmoscopically invisible lesions could be detected and lesions that were under- or overexposed could be distinguished using the OCT data.

The objective of this work is to assess oxidative stress levels in salt-sensitive hypertension animal model using 3D optical cryoimager to image mitochondrial redox ratio. We studied Dahl salt-induced (SS) rats, and compared the results with a consomic SS rat strain (SSBN13). The SSBN13 strain was developed by the introgression of chromosome from the Brown Norway (BN) rat into the salt-sensitive (SS) genetic background and exhibits significant protection from salt induced hypertension¹ . These two groups were fed on a high salt diet of 8.0% NaCl for one week. Mitochondrial redox ratio (NADH/FAD=NADH RR), was used as a quantitative marker of the oxidative stress in kidney tissue. Maximum intensity projected images and their corresponding histograms in each group were acquired from each kidney group. The result showed a 49% decrease in mitochondrial redox ratio of SS compared to SSBN13 translated to an increase in the level of oxidative stress of the tissue. Therefore, the results quantify oxidative stress levels and its effect on mitochondrial redox in salt sensitive hypertension.

Low-level therapy laser is a phototherapy treatment that involves the application of low power light in the red or infrared wavelengths in various diseases such as arthritis. In this work, we investigated whether low-intensity infrared laser therapy could cause death by caspase-6 apoptosis or DNA damage pathways in cartilage cells after zymosaninduced articular inflammatory process. Inflammatory process was induced in C57BL/6 mouse by intra-articular injection of zymosan into rear tibio-tarsal joints. Thirty animals were divided in five groups: (I) control, (II) laser, (III) zymosan-induced, (IV) zymosan-induced + laser and (V). Laser exposure was performed after zymosan administration with low-intensity infrared laser (830 nm), power 10 mW, fluence 3.0 J/cm² at continuous mode emission, in five doses. Twenty-four hours after last irradiation, the animals were sacrificed and the right joints fixed and demineralized. Morphological analysis was observed by hematoxylin and eosin stain, pro-apoptotic (caspase-6) was analyzed by immunocytochemistry and DNA fragmentation was performed by TUNEL assay in articular cartilage cells. Inflammatory process was observed in connective tissue near to articular cartilage, in IV and V groups, indicating zymosan effect. This process was decreased in both groups after laser treatment and dexamethasone. Although groups III and IV presented higher caspase-6 and DNA fragmentation percentages, statistical differences were not observed when compared to groups I and II. Our results suggest that therapies based on low-intensity infrared lasers could reduce inflammatory process and could not cause death by caspase-6 apoptosis or DNA damage pathways in cartilage cells after zymosan-induced articular inflammatory process.

In order to develop minimally invasive, fast and precise diagnostic and therapeutic methods in medicine by using optical methods, first step is to examine how the light propagates, scatters and transmitted through medium. So as to find out appropriate wavelengths, it is required to correctly determine the optical properties of tissues. The aim of this study is to measure the optical properties of both cancerous and normal ex-vivo pancreatic tissues. Results will be compared to detect how cancerous and normal tissues respond to different wavelengths. Double-integrating-sphere system and computational technique inverse adding doubling method (IAD) were used in the study. Absorption and reduced scattering coefficients of normal and cancerous pancreatic tissues have been measured within the range of 500-650 nm. Statistical significant differences between cancerous and normal tissues have been obtained at 550 nm and 630 nm for absorption coefficients. On the other hand; there were no statistical difference found for scattering coefficients at any wavelength.