This PDF file contains the front matter associated with SPIE Proceedings Volume 9706, including the Title Page, Copyright information, Table of Contents, Introduction, and the Conference Committee listing.

This paper evaluates image enhancement and visualization techniques for pulsed terahertz (THz) images of tissue samples. Specifically, our research objective is to effectively differentiate between heterogeneous regions of breast tissues that contain tumors diagnosed as triple negative infiltrating ductal carcinoma (IDC). Tissue slices and blocks of varying thicknesses were prepared and scanned using our lab’s THz pulsed imaging system. One of the challenges we have encountered in visualizing the obtained images and differentiating between healthy and cancerous regions of the tissues is that most THz images have a low level of details and narrow contrast, making it difficult to accurately identify and visualize the margins around the IDC. To overcome this problem, we have applied and evaluated a number of image processing techniques to the scanned 3D THz images. In particular, we employed various spatial filtering and intensity transformation techniques to emphasize the small details in the images and adjust the image contrast. For each of these methods, we investigated how varying filter sizes and parameters affect the amount of enhancement applied to the images. Our experimentation shows that several image processing techniques are effective in producing THz images of breast tissue samples that contain distinguishable details, making further segmentation of the different image regions promising.

The ability to discern malignant from benign tissue in excised human breast specimens in Breast Conservation Surgery (BCS) was evaluated using a prototype single frequency terahertz radiation. Terahertz (THz) images of the specimens in reflection mode were obtained by employing a gas laser source and mechanical scanning. The images were correlated with optical histological micrographs of the same specimens, and a mean discrimination of 73% was found for five out of six samples using Receiver Operating Characteristic (ROC) analysis. This result is similar to what has previously been obtained using Terahertz pulsed imaging (TPI) techniques. We will discuss the specific advantages of Single frequency THz imaging (SFTI) compared with TPI for potentially allowing the development of much faster, more compact and less expensive cancer imaging systems that could be adapted for employment in the operating room. The system design and characterization of the prototype SFTI system are discussed in detail. The initial results are encouraging but further development of the technology and clinical evaluation is needed to evaluate its feasibility in the clinical environment.

In this presentation, a review and quasioptical imaging system and design considerations for an off-axis parabolic mirror based THz imaging systems are presented. It is shown that off-axis parabolic mirrors introduce off-axis intensity and polarization distortion. When a train of OAPs are used to relay THz beam, each distortions rapidly stacks to produce quite ugly beam and polarization profile. We show that the distortion of field distribution and polarization as a function of mirror curvature and focusing parameters. A brief review of design rules are shown to eliminate these distortions by a symmetric configurations of off-axis parabolic mirror train. The detrimental distortion effects were cancelled out by orienting the final two mirrors in a way to that recovers the original source profile. Comparisons of field profiles between compensated and uncompensated design are shown and imaging performance on characterization targets presented. In addition to benefits in field and polarization distribution the improved design facilitates 1D scanning with minimal change to overall optical path length.

The THz electromagnetic properties of rough surface are explored and their effect on the observed contrast in THz images is quantified. Rough surface scatter is a major source of clutter in THz imaging as the rough features of skin and other tissues result in non-trivial reflection signal modulation. Traditional approaches to data collection utilize dielectric windows to flatten surfaces for THz imaging. However, there is substantial interest surrounding window free imaging as contact measurements are not ideal for a range of candidate diseases and injuries. In this work we investigate the variation in reflected signal in the specular direction from rough surfaces targets with known roughness parameters. Signal to clutter ratios are computed and compared with that predicted by Rayleigh Rough surface scattering theory. It is shown that Rayleigh rough surface scattering theory, developed for rough features larger than the interacting wavelength, holds acceptable at THz frequencies with rough features much smaller than the wavelength. Additionally, we present some biological tissue imaging examples to illustrate the impact of rough surface scattering in image quality.

Recently, some studies reported that the sweat ducts act as a low-Q-factor helical antenna due to their helical structure, and resonate in the terahertz frequency range according to their structural parameters. According to the antenna theory, when the duct works as a helical antenna, the dimension of the helix plays a key role to determine the frequency of resonance. Therefore, the accurate determination of structural parameters of sweat duct is crucially important to obtain the reliable frequency of resonance and modes of operations. Therefore, here we performed the optical coherence tomography (OCT) of human subjects on their palm and foot to investigate the density, distribution and morphological features of sweat ducts. Moreover, we measured the dielectric properties of stratum corneum using terahertz time domain spectroscopy and based upon this information, we determined the frequency of resonance. We recruited 32 subjects for the measurement and the average duct diameter was 95±11μm. Based upon this information on diameter of duct and THz dielectric properties of stratum corneum (ε=5.1±1.3), we have calculated the frequency of resonance of sweat duct. Finally, we determined that the center frequency of resonance was 442±76 GHz. We believe that these findings will facilitate further investigation of the THz-skin interaction and provide guidelines for safety levels with respect to human exposure. We will also report on the EEG measurement while being shined by micro watt order THz waves.

This paper explores the utility of reflective THz imaging to assess the viability of surgical flaps. Flap surgery is a technique where tissue is harvested from a donor site and moved to a recipient while keeping the blood supply intact. This technique is common in head and neck tumor resection surgery where the reconstruction of complex and sensitive anatomic structures is routine following the resection of large and/or invasive tumors. Successful flap surgery results in tissue that is sufficiently perfused with both blood and extracellular water. If insufficient fluid levels are maintained, the flap tissue becomes necrotic and must be excised immediately to prevent infection developing and spreading to the surrounding areas. The goal of this work is to investigate the hydration of surgical flaps and correlate image features to successful graft outcomes. Advancement flaps were created on the abdomens of rat models. One rat model was labeled control and care was taken to ensure a successful flap outcome. The flap on the second rat was compromised with restricted blood flow and allowed to fail. The flaps of both rats were imaged once a day over the course of a week at which point the compromised flap had begun to show signs of necrosis. Significant differences in tissue water content were observed between rats over the experimental period. The results suggest that THz imaging may enable early assessment of flap viability.

Well-regulated corneal water content is critical for ocular health and function and can be adversely affected by a number of diseases and injuries. Current clinical practice limits detection of unhealthy corneal water content levels to central corneal thickness measurements performed by ultrasound or optical coherence tomography. Trends revealing increasing or decreasing corneal thickness are fair indicators of corneal water content by individual measurements are highly inaccurate due to the poorly understood relationship between corneal thickness and natural physiologic variation. Recently the utility of THz imaging to accuarately measure corneal water content has been explored on with rabbit models. Preliminary experiments revealed that contact with dielectric windows confounded imaging data and made it nearly impossible to deconvolve thickness variations due to contact from thickness variations due to water content variation. A follow up study with a new optical design allowed the acquisition of rabbit data and the results suggest that the observed, time varying contrast was due entirely to the water dynamics of the cornea. This paper presents the first ever in vivo images of human cornea. Five volunteers with healthy cornea were recruited and their eyes were imaged three times over the course of a few minutes with our novel imaging system. Noticeable changes in corneal reflectivity were observed and attributed to the drying of the tear film. The results suggest that clinically compatible, non-contact corneal imaging is feasible and indicate that signal acquired from non-contact imaging of the cornea is a complicated coupling of stromal water content and tear film.

This paper explores vasodynamics in response to histamine injection using reflective THz imaging. Histamine is a major contributor to allergic disease. Elevations in tissue histamine levels have been observed during anaphylaxis and experimental allergic responses of the skin, nose, and airways. In the skin specifically, vasodilation, vascular permeability, and pruritus is controlled by the release and resorption of histamine. These properties are leveraged in skin prick testing for allergies where histamine dihydrochloride is injected as a positive control to confirm allergen susceptibility prior to the administration of candidate allergens. Subjective parameters such as skin coloration, irritation, and bulging as a consequence of histamine injection and histamine release are well characterized. However limited quantitative metrics on the body’s edematous response are available due to the lack of imaging diagnostics that can map surface tissue water content (TWC). THz imaging was used to explore the utility of reflective THz imaging to quantify edematous responses to histamine. Rat models were injected with varying concentrations of histamine dihydrochloride and the resultant edematous response arising from perturbed vasodymanics was mapped. Significant build up and dissipation of surface tissue water content was observed and THz frequency contrast was seen to correlate with visual appearance in some cases and in others reveal tissue water content variations not discernable with the naked eye. The results suggest that THz imaging may be a valuable tool in quantifying the degree of allergic responses and assist in detecting hypersensitivity.

During laparoscopic surgery, devices are require to either cut, ablate or coagulate tissue and veins with high precision and controlled lateral damage preferably in an one-for-all modality. The tissue interactions of 3 new treatment modalities were studied using special imaging techniques to obtain a better understanding the working mechanism in view of effective and safe application. The Plasmajet produces a high temperature ionized gas 'flame' directed to the tissue surface at the tip of a 4 mm diameter rigid hand piece. The Lumenis DUO CO2 laser enables endoscopic laser energy delivery through a 1 mm outer diameter flexible hollow waveguide. The 2 µm 'Thulium' laser is delivered by (standard) 400 µm diameter optical fiber. Thermal imaging and Schlieren techniques were used to assess the superficial ablative and coagulation effects these surgical instruments scanning at preset velocities and distances from the surface of biological tissues and phantoms . The CO2 was very effective in tissue ablation even at a distance up to 10 mm due to a very small diverging beam from the hollow waveguide. In contrast, the Thulium laser showed less ablation and increasing coagulation at larger distance to the tissue. The gas 'flame' of the Plasmajet spread the thermal energy over the surface for effective superficial ablation and coagulation. However, the pressure of the gas flow is substantial on the tissue surface creating turbulence and even indirect cooling. The specific ablation and coagulation effects of the three treatment modalities have to be appreciate and the effective and safe application will depend on the preference and skills of the surgeon

This paper describes the use of a 420 nm blue laser diode for possible surgery and hemostasis. The optical absorption of blood-containing tissue is strongly determined by the absorption characteristics of blood. Blood is primarily comprised of plasma (yellowish extracellular fluid that is approximately 95% water by volume) and formed elements: red blood cells (RBCs), white blood cells (WBCs) and platelets. The RBCs (hemoglobin) are the most numerous, and due to the spectral absorption characteristics of hemoglobin, the optical absorption of blood has a strong relative maximum value in the 420 nm blue region of the optical spectrum. Small, low-cost laser diodes emitting at 420 nm with tens of watts of continuous wave (CW) optical power are becoming commercially available. Experiments on the use of such laser diodes for tissue cutting with simultaneous hemostasis were carried out and are here described. It was found that 1 mm deep x 1 mm wide cuts can be achieved in red meat at a focused laser power level of 3 W moving at a velocity of ~ 1 mm/s. The peripheral necrosis and thermal damage zone extended over a width of approximately 0.5 mm adjacent to the cuts. Preliminary hemostasis experiments were carried out with fresh equine blood in Tygon tubing, where it was demonstrated that cauterization can occur in regions of intentional partial tubing puncture.

Protein therapeutics have been developed to treat diseases ranging from arthritis and psoriasis to cancer. A challenge in the development of protein-based drugs is maintaining the protein in the folded state during processing and storage. We are developing a novel processing method, light-assisted drying (LAD), to dehydrate proteins suspended in a sugar (trehalose) solution for storage at supra-zero temperatures. Our technique selectively heats the water in small volume samples using near-IR light to speed dehydration which prevents sugar crystallization that can damage embedded proteins. In this study, we compare the end moisture content (EMC) as a function of processing time of samples dried with two different light sources, Nd:YAG (1064 nm) and Thulium fiber (1850 nm) lasers. EMC is the ratio of water to dry weight in a sample and the lower the EMC the higher the possible storage temperature. LAD with the 1064 and 1850 nm lasers yielded 78% and 65% lower EMC, respectively, than standard air-drying. After 40 minutes of LAD with 1064 and 1850 nm sources, EMCs of 0.27±.27 and 0.15±.05 gH₂O/gDryWeight were reached, which are near the desired value of 0.10 gH₂O/gDryWeight that enables storage in a glassy state without refrigeration. LAD is a promising new technique for the preparation of biologics for anhydrous preservation.

We studied heating drug delivery to vascular wall with Rhodamine B ranging 50 to 70°C ex vivo study. Porcine carotid artery was dipped in the heated Rhodamine B solution in 15 s and then cooled by 37°C saline. Rhodamine B concentration distribution in the vascular wall cross-section was measured by a fluorescence microscope using 550 nm for excitation and 620 nm emission for fluorescence detection. The total amount of measured fluorescence in the vascular wall was calculated as a indication of delivered Rhodamine B quantity. The delivered Rhodamine B quantity was increased with increasing heating temperature with 50 to 70°C. In the cases of 60 to 70°C heating, the delivered Rhodamine B quantity was 3.1 to 23.3 fold by that of 37°C. Defined penetration depth of the delivered Rhodamine B in the vascular wall was also significantly increased with 65°C and 70°C heating. We also studied heating drug delivery to the vascular wall with fluorescence labeled Paclitaxel with 70°C in 15 s and 60 s heating ex vivo. In both contact duration, the delivered Paclitaxel quantity was increased. To understand these drug delivery enhancement effects, we investigated the vascular cross-sectional structure change by the heating. Some holes over 50 nm in diameter appeared on the internal elastic lamina with 70°C heating. We prospected that vascular surface structure change by the heating might enhance drug delivery to the vascular wall.

We have proposed to apply the photosensitization reaction in myocardium interstitial fluid using talaporfin sodium to realize less-heated electrical conduction block for a tachyarrhythmia treatment: PD Ablation^®. The cytotoxicity of the extracellular photosensitization reaction efficiency may change by the talaporfin sodium binding with serum proteins. These binding would change with solution temperature. We investigated the binding behavior of talaporfin sodium with human serum albumin (HSA), high-density lipoprotein (HDL), and low-density lipoprotein (LDL) changing solution temperature from 17 to 37°C. We also studied the photocytotoxicity change by solution temperature of 17 and 37°C measuring cell lethality by WST assay using fetal bovine serum. The binding ratio of talaporfin sodium with HDL and LDL decreased 6.3% and 12.8% with temperature increasing from 17 to 37°C. There was no significant difference in the case of HSA. The cell lethality was increased about 30% with temperature increasing from 17 to 37°C. The myocardium tissue temperature increase was reported that less than 5°C in the case of our PD Ablation^®. We think that the photocytotoxicity change by these temperature increasing would be negligible in our PD Ablation^®. We suggest that the temperature maintaining would be necessary to keep the photocytotoxicity efficiency in the case of the open surgery that would cause the tissue surface temperature decreasing.

We report on development of optical parametric oscillator (OPO) based mid-infrared laser system, which utilizes periodically poled nonlinear crystal that was pumped by near-infrared (NIR) laser. We have obtained 8 W of mid-infrared average output at the injection current of 20A from a quasi-phase-matched OPO using external cavity configuration. The laser tissue ablation efficiency was investigated which is substantially affected by several parameters such as, optical fluence rate, wavelength of the laser source and the optical properties of target tissue. Wavelength and radiant exposure dependent tissue ablation dimension were quantified by using SD-OCT (spectral domain optical coherence tomography) and the ablation efficiency was compared to that of non-converted NIR- laser system.

Cholesteryl esters are main components of atherosclerotic plaques and have an absorption peak at the wavelength of 5.75 μm originated from C=O stretching vibration mode of ester bond. Our group achieved the selective ablation of atherosclerotic lesions 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 considered to be useful for clinical application. However, large thermal effects were observed because the QCL worked as quasicontinuous wave (CW) lasers due to its short pulse interval. Then we tried macro pulse irradiation (irradiation of pulses at intervals) of the QCL and achieved effective ablation with less-thermal effects than conventional quasi-CW irradiation. However, lesion selectivity might be changed by changing pulse structure. Therefore, in this study, irradiation effects of the macro pulse irradiation to rabbit atherosclerotic plaque and normal vessel were compared. The macro pulse width and the macro pulse interval were set to 0.5 and 12 ms, respectively, because the thermal relaxation time of rabbit normal and atherosclerotic aortas in the oscillation wavelength of the QCL was 0.5–12 ms. As a result, cutting difference was achieved between rabbit atherosclerotic and normal aortas by the macro pulse irradiation. Therefore, macro pulse irradiation of a QCL in the 5.7 μm wavelength range is effective for reducing thermal effects and selective ablation of the atherosclerotic plaque. QCLs have the potential of realizing less-invasive laser angioplasty.

Photothermal medical laser treatments are extremely dependent on the generated tissue temperature. It is necessary to reach a certain temperature threshold to achieve successful results, whereas preventing to exceed an upper temperature value is required to avoid thermal damage. One method to overcome this problem is to use previously conducted dosimetry studies as a reference. Nevertheless, these results are acquired in controlled environments using uniform subjects. In the clinical environment, the optical and thermal characteristics (tissue color, composition and hydration level) vary dramatically among different patients. Therefore, the most reliable solution is to use a closed-loop feedback system that monitors the target tissue temperature to control laser exposure. In this study, we present a compact, non-contact temperature measurement system for the control of photothermal medical laser applications that is cost-efficient and simple to use. The temperature measurement is achieved using a focused, commercially available MOEMS infrared thermocouple sensor embedded in an off-axis arrangement on the laser beam delivery hand probe. The spot size of the temperature sensor is ca. 2.5 mm, reasonably smaller than the laser spot sizes used in photothermal medical laser applications. The temperature readout and laser control is realized using a microcontroller for fast operation. The utilization of the developed system may enable the adaptation of several medical laser treatments that are currently conducted only in controlled laboratory environments into the clinic. Laser tissue welding and cartilage reshaping are two of the techniques that are limited to laboratory research at the moment. This system will also ensure the safety and success of laser treatments aiming hyperthermia, coagulation and ablation, as well as LLLT and PDT.

In this study, we present a fast analytical approach for laser induced temperature increase in biological tissue. The whole problem consists of two main steps. These steps are the light propagation and heat transfer in tissue. We first obtain a detailed analytical solution for the diffusion equation based on an integral approach for specific boundary conditions. Secondly, we also solve the Pennes' bio-heat transfer equation analytically using the separation of variables technique and obtain the temperature induced by optical absorption of tissue. Here, heat source term consists of the local absorption and photon density, which will be determined from the diffusion equation. We find a very comprehensive solution for the diffusion equation by using an integral method for the Robin boundary condition. In other words, we obtain a particular Green's function in a different way. Next, we use this solution as a source term in the Pennes’ bio-heat equation by utilizing the heat convection boundary condition. It is important to note that these boundary conditions are good approximations for imaging of biological tissue. As a result, we obtain spatio-temporal temperature distribution inside the medium. First, our approach is validated by a numerical approach using a Finite Element Method (FEM). Next, we also validate our method by performing phantom and tissue experiments. Experimental data corresponding to spatio-temporal temperature distribution are recorded using magnetic resonance thermometry. The analytical results obtained by our method are in a very good agreement with ones obtained by the FEM and experiment.

One major advantage of using gold nanoparticles is the possibility of tuning their absorption peak by modifying their surface plasma resonance. They are proven to be a promising multi-functional platform that can be used for many imaging and therapeutic applications. As a true multi-modality imaging technique, Photo-Magnetic Imaging (PMI) has a great potential to monitor the distribution of gold nanoparticles non-invasively with MR resolution. With a simple addon of a continuous wave laser to an MRI system, PMI uses the laser induced temperature increase, measured by MR Thermometry (MRT), to provide tissue optical absorption maps at MR resolution. PMI utilizes a Finite Element Method (FEM) based algorithm to solve the combined diffusion and bio-heat equations. This system of combined equations models the photon distribution in the tissue and heat generation due to the absorption of the light and consequent heat diffusion. The key characteristic of PMI is that its spatial resolution is preserved at any depth as long as the temperature change within the imaged medium is detectable by MRT. Agar phantoms containing gold nanoparticles are used to validate the ability of PMI in monitoring their distribution. To make PMI suitable for diagnostic purposes, the laser powers has been kept under the American National Standard Institute maximum skin exposure limits in this study.

Glaucoma is the second-leading cause of blindness worldwide and is often associated with elevated intraocular pressure (IOP). Partial-thickness drainage channels can be created with femtosecond laser in the translucent sclera for the potential treatment of glaucoma. We demonstrate the creation of partial-thickness subsurface drainage channels with the femtosecond laser in the cadaver human eyeballs and describe the application of two-photon microscopy and confocal microscopy for noninvasive imaging of the femtosecond laser created partial-thickness scleral channels in cadaver human eyes. A femtosecond laser operating at a wavelength of 1700 nm was scanned along a rectangular raster pattern to create the partial thickness subsurface drainage channels in the sclera of cadaver human eyes. Analysis of the dimensions and location of these channels is important in understanding their effects. We describe the application of two-photon microscopy and confocal microscopy for noninvasive imaging of the femtosecond laser created partial-thickness scleral channels in cadaver human eyes. High-resolution images, hundreds of microns deep in the sclera, were obtained to allow determination of the shape and dimension of such partial thickness subsurface scleral channels. Our studies suggest that the confocal and two-photon microscopy can be used to investigate femtosecond-laser created partial-thickness drainage channels in the sclera of cadaver human eyes.

Experimental results of femtosecond Laser Assisted Bioprinting (LAB) are reported on. Two set-up, used to print different model bioinks and keratinocytes cells line HaCaT, were studied: first one was using a femtosecond laser with low pulse energy and an absorbing gold layer, whereas the second one used high pulse energy enabling the removal of the absorbing layer. Printed drop diameter and resulting height of the bioink jet are then quantified as a function of the LAB parameters such as laser energy, focus spot location or numerical aperture.

Endoscopic resection of early colorectal neoplasms typically employs electrocautery tools, which lack precision and run the risk of full thickness thermal injury to the bowel wall with subsequent perforation. We present a means of endoluminal colonic ablation using picosecond laser pulses as a potential alternative to mitigate these limitations. High intensity ultrashort laser pulses enable nonlinear absorption processes, plasma generation, and as a consequence a predominantly non-thermal ablation regimen.

Robust process parameters for the laser resection are demonstrated using fresh ex vivo pig intestine samples. Square cavities with comparable thickness to early colorectal neoplasms are removed for a wavelength of 1030 nm and 515 nm using a picosecond laser system. The corresponding histology sections exhibit in both cases only minimal collateral damage to the surrounding tissue. The ablation depth can be controlled precisely by means of the pulse energy. Overall, the application of ultrafast lasers for the resection of intestine enables significantly improved precision and reduced thermal damage to the surrounding tissue compared to conventional electrocautery.

Femtosecond laser oscillator systems with low pulse energy (< 1 μJ) and high repetition rate (MHz) are increasingly used for precise, fast and safe eye surgery. Therefore, the laser tissue interaction process is of great interest to optimize and improve established and future surgical protocols. Besides, using faster laser systems leads to unintended self-induced interaction effects, where a femtosecond laser pulse modifies the vicinity in the material in such a way that the focus of following laser pulses is changed. We used a femtosecond oscillator laser system with high repetition rate and 66 nJ pulse energy to produce photodisruption in water. Water was used as phantom material for ocular tissue, because tissue mainly consists of water. A custom made digital-holographic system was used to measure the temporal material modification from picoseconds until seconds after occurrence of the photodisruption. For illumination of the sample we used either a continuously light source or the femtosecond laser pulse itself in a pump-probe configuration. The holographic system provides quantitative data of phase difference Δφ for the full field of view of several tenth of micrometers. Phase difference is equivalent to the laser induced change in the material’s refractive index which can alter focusing conditions of following laser pulses and might impair surgical outcome. We obtained the largest change in Δφ during the first picoseconds, followed by a slow relaxation of Δφ within some milliseconds. The results of time resolved measurements of the laser induced material modification will help to optimize scanning schemes in ocular surgery.

The interaction effect of photodisruption, which is used for dissection of biological tissue with fs-laser pulses, has been intensively studied inside water as prevalent sample medium. In this case, the single effect is highly reproducible and, hence, the method of time-resolved photography is sufficiently applicable. In contrast, the reproducibility significantly decreases analyzing more solid and anisotropic media like biological tissue. Therefore, a high-speed photographic approach is necessary in this case. The presented study introduces a novel technique for high-speed photography based on the principle of chromatic encoding. For illumination of the region of interest within the sample medium, the light paths of up to 12 LEDs with various emission wavelengths are overlaid via optical filters. Here, MOSFET-electronics provide a LED flash with a duration <100 ns; the diodes are externally triggered with a distinct delay for every LED. Furthermore, the different illumination wavelengths are chromatically separated again for detection via camera chip. Thus, the experimental setup enables the generation of a time-sequence of ≤ 12 images of a single cavitation bubble dynamics. In comparison to conventional time-resolved photography, images in sample media like water and HEMA show the significant advantages of this novel illumination technique. In conclusion, the results of this study are of great importance for the fundamental evaluation of the laser-tissue interaction inside anisotropic biological tissue and for the optimization of the surgical process with high-repetition rate fs-lasers. Additionally, this application is also suitable for the investigation of other microscopic, ultra-fast events in transparent inhomogeneous materials.

Lead extraction (LE) is necessary for patients who are suffering from a related infection, or in opening venous occlusions that prevent the insertion of additional lead. In severe cases of fibrous encapsulation of the lead within a vein, laser-based cardiac LE has become one of the foremost methods of removal. In cases where the laser radiation (typically at 308 nm wavelength) interacts with the vein wall rather than with the fibrotic lesion, severe injury and subsequent bleeding may occur. Selective tissue ablation was previously demonstrated by a laser operating in the UV regime; however, it requires the use of sensitizers (e.g.: tetracycline). In this study, we present a preliminary examination of efficacy and safety aspects in the use of a nanosecond-pulsed solid-state laser radiation, at 355 nm wavelength, guided in a catheter consisting of optical fibers, in LE. Specifically, we demonstrate a correlation between the tissue elasticity and the catheter advancement rate, in ex-vivo experiments. Our results indicate a selectivity property for specific parameters of the laser radiation and catheter design. The selectivity is attributed to differences in the mechanical properties of the fibrotic tissue and a normal vein wall, leading to a different photomechanical response of the tissue’s extracellular matrix. Furthermore, we performed successful in-vivo animal trials, providing a basic proof of concept for using the suggested scheme in LE. Selective operation using a 355 nm laser may reduce the risk of blood vessel perforation as well as the incidence of major adverse events.

The Photonic Fence is a system designed to detect mosquitoes and other pestilent flying insects in an active region and to apply lethal doses of laser light to them. Previously, we determined lethal fluence levels for a variety of lasers and pulse conditions on anesthetized Anopheles stephensi mosquitoes. In this work, similar studies were performed while the bugs were freely flying within transparent cages. Dose-response curves were created for various beam diameter, pulse width, and power conditions at 455 nm, 532 nm, 1064nm, and 1540 nm wavelengths. Besides mortality outcomes, the flight behavior of the bugs and the performance of the tracking system were monitored for consistency and to ensure that they had no impact on the mortality outcomes. As in anesthetized experiments, the visible wavelengths required significantly less fluence than near infrared wavelengths to reliably disable bugs. For the visible wavelengths, lethal fluence values were generally equivalent to those found in anesthetized dosing, while near infrared wavelengths required approximately twice the fluence compared with anesthetized experiments. The performance of the optical tracking system remained highly stable throughout the experiments, and it was found not to influence mortality results for pulse widths up to 25 ms. In general, keeping energy constant while decreasing power and increasing pulse width reduced mortality levels. The results of this study further affirm the practicality of using optical approaches to protect people and crops from flying insects.

Nanoporation occurs in cells exposed to high amplitude short duration (< 1μs) electrical pulses. The biophysical mechanism(s) responsible for nanoporation is unknown although several theories exist. Current theories focus exclusively on the electrical field, citing electrostriction, water dipole alignment and/or electrodeformation as the primary mechanisms for pore formation. Our group has shown that mechanical forces of substantial magnitude are also generated during nsEP exposures. We hypothesize that these mechanical forces may contribute to pore formation. In this paper, we report that alteration of the conductivity of the exposure solution also altered the level of mechanical forces generated during a nsEP exposure. By reducing the conductivity of the exposure solutions, we found that we could completely eliminate any pressure transients normally created by nsEP exposure. The data collected for this proceeding does not definitively show that the pressure transients previously identified contribute to nanoporation; however; it indicates that conductivity influences both survival and pressure transient formation.

Cells exposed to nanosecond-pulsed electric fields (nsPEF) exhibit a wide variety of nonspecific effects, including blebbing, swelling, intracellular calcium bursts, apoptotic and necrotic cell death, formation of nanopores, and depletion of phosphatidylinositol 4,5-biphosphate (PIP2) to induce activation of the inositol trisphosphate/diacylglycerol pathway. While several studies have taken place in which multiple pulses were delivered to cells, the effect of pulse repetition rate (PRR) is not well understood. To better understand the effects of PRR, a laser scanning confocal microscope was used to observe CHO-K1 cells exposed to ten 600ns, 200V pulses at varying repetition rates (5Hz up to 500KHz) in the presence of either FM 1-43, YO-PRO-1, or Propidium Iodide (PI) fluorescent dyes, probes frequently used to indicate nanoporation or permeabilization of the plasma membrane. Dye uptake was monitored for 30 seconds after pulse application at a rate of 1 image/second. In addition, a single long pulse of equivalent energy (200V, 6 μs duration) was applied to test the hypothesis that very fast PRR will approximate the biological effects of a single long pulse of equal energy. Upon examination of the data, we found strong variation in the relationship between PRR and uptake in each of the three dyes. In particular, PI uptake showed little frequency dependence, FM 1-43 showed a strong inverse relationship between frequency and internal cell fluorescence, and YO-PRO-1 exhibited a “threshold” point of around 50 KHz, after which the inverse trend observed in FM 1-43 was seen to reverse itself. Further, a very high PRR of 500 KHz only approximated the biological effects of a single 6 μs pulse in cells stained with YO-PRO-1, suggesting that uptake of different dyes may proceed by different physical mechanisms.

Polarization-sensitive optical coherence tomography (PS-OCT) allows imaging of tissue birefringence. In practice, however, PS-OCT images are often confounded by high noise and confusing artifacts. A full understanding of the intrinsic and instrumentation-derived signal and noise properties of PS-OCT has not been developed. In this presentation, we describe a Monte Carlo (MC) simulator of PS-OCT local birefringence imaging that recapitulates the noise and signal properties observed in empirical images and, as such, can be used to understand and improve PS-OCT methods. The MC simulator builds upon a previously described MC methodology that supports arbitrary three-dimensional geometries. To this, we have added support for MC simulation of transverse speckle correlation. This is important because many of the noise sources in PS-OCT are driven by interactions with the speckle field. We have developed a method to support polarization-dependent measurements of birefringent tissues. Both additive (due to finite SNR) and polarization-mode dispersion noise can be incorporated. To demonstrate the utility of the simulator, we use it to reveal a previously unappreciated noise that results from the design of conventional local birefringence extraction algorithms, and we describe an improved method that lowers noise in both simulated and empirical datasets. We anticipate that this simulator will enable new explorations into the fidelity of PS-OCT measurements and accelerate the optimization of PS-OCT methods and algorithms.

Gastrointestinal tumoral pathologies are quite common nowadays. Diseases such as gastric antral vascular ectasia (GAVE) or actinic proctitis may require endoscopic surgery. Argon Plasma Coagulated (APC) or radiofrequency are usually employed. However, they present disadvantages, such as the reduced treated area, magnetic resonance incompatibility, or an uncontrolled ablation depth. Optical surgery could avoid these problems and contribute to a better and controlled treatment result, either ablative or coagulative, in a minimally invasive, non-contact and non-ionizing way. The treatment area could also be increased by adequate optical fiber probe design. In this work laser surgery is analyzed for resection of colonic tumors. A Monte Carlo model is employed to study optical propagation, and an optical ablation approach allows the estimation of the resected volume. The ablation approach is based on plasma-induced ablation, particularly taking into account the freeelectron density generated in the tissue by the pulsed optical source. Several wavelengths, radii and malignant tissue types are considered, either healthy, adenomatous or even coagulated tissues. Optimum source parameters as a function of tumor geometry can be estimated for treatment planning.

In simulations of light transport in biological tissues and organs knowledge of tissue optical properties is imperative for realism of the predicted effects. One factor which is commonly overlooked is the choice of appropriate scattering phase function. Henyey-Greenstein phase function (PF) is often applied due to its suitability for analytical derivations and availability of the corresponding tissue anisotropy factors. At the same time it is known that it doesn't match the angular distribution of scattering in many tissues. In here, we study the influence of the PF in 3D Monte Carlo simulations of hyperspectral imaging (HSI). For a simple geometrical (three-layered) model of skin and a discrete blood vessel, hyperspectral images in the 400–1000 nm spectral range were simulated using Henyey-Greenstein, modified Henyey-Greenstein, and Mie PF, respectively. The results are compared in the spatial and spectral domains. In addition, the effective tissue properties as determined from the simulated HSI using 1D inverse MC are compared with the input parameter values. The results show that the choice of PF assumed in light transport models has a substantial impact on simulated HSI. Using an inappropriate PF can result in significantly altered HSI and considerable artifacts in extracted values of the skin parameters.

Optogenetics provides a tool for modulating activity of specific cell types by light pulses. Different light delivery mechanisms such as single optical fiber implanted on a skull or patterned illumination can be employed to direct light to a target area. For a highly scattering medium such as brain tissue, light distribution significantly depends on the scattering parameters of the tissue as well as the inherent inhomogeneity of the specimen. For in vivo studies, blood vessels which have considerable absorption coefficient in the visible spectrum play a major role in producing such inhomogeneity. Therefore, detailed information about brain optical properties and network of blood vessels which was ignored in previous studies is necessary to accurately predict light distribution and designing light delivery mechanism during optogenetic experiments to achieve the desired optical stimulation. In this paper, light pattern preservation while considering the impact of blood vessels is investigated in a rat cortex. First, the typical optical properties of rat cortical tissue were extracted by employing double integrated sphere technique, and then, optical coherence tomography was employed to obtain structure of blood vessels on the cortex. By combining the extracted optical properties and the network of blood vessels, a three-dimensional model of a rat cortical tissue was developed. Then, a Monte Carlo simulation code was used to predict light distribution in this model for different light source configurations and wavelengths. The results confirm that presence of vessels can significantly impact the light pattern in the tissue and affect the practical depth of penetration.

Simulating propagation and scattering of coherent light in turbid media, such as biological tissues, is a complex problem. Numerical methods for solving Helmholtz or wave equation (e.g. finite-difference or finite-element methods) require large amount of computer memory and long computation time. This makes them impractical for simulating laser beam propagation into deep layers of tissue. Other group of methods, based on radiative transfer equation, allows to simulate only propagation of light averaged over the ensemble of turbid medium realizations. This makes them unuseful for simulating phenomena connected to coherence properties of light. We propose a new method for simulating propagation of coherent light (e.g. laser beam) in biological tissue, that we called Coherent-Wave Monte Carlo method. This method is based on direct computation of optical interaction between scatterers inside the random medium, what allows to reduce amount of memory and computation time required for simulation. We present the theoretical basis of the proposed method and its comparison with finite-difference methods for simulating light propagation in scattering media in Rayleigh approximation regime.

Monte Carlo simulations are widely considered to be the gold standard for studying the propagation of light in turbid media. However, due to the probabilistic nature of these simulations, large numbers of photons are often required in order to generate relevant results. Here, we present methods for reduction in the variance of dose distribution in a computational volume. Dose distribution is computed via tracing of a large number of rays, and tracking the absorption and scattering of the rays within discrete voxels that comprise the volume. Variance reduction is shown here using quasi-random sampling, interaction forcing for weakly scattering media, and dose smoothing via bi-lateral filtering. These methods, along with the corresponding performance enhancements are detailed here.

Monte Carlo simulations are widely considered to be the gold standard for studying the propagation of light in turbid media. However, traditional Monte Carlo methods fail to account for diffraction because they treat light as a particle. This results in converging beams focusing to a point instead of a diffraction limited spot, greatly effecting the accuracy of Monte Carlo simulations near the focal plane. Here, we present a technique capable of simulating a focusing beam in accordance to the rules of Gaussian optics, resulting in a diffraction limited focal spot. This technique can be easily implemented into any traditional Monte Carlo simulation allowing existing models to be converted to include accurate focusing geometries with minimal effort. We will present results for a focusing beam in a layered tissue model, demonstrating that for different scenarios the region of highest intensity, thus the greatest heating, can change from the surface to the focus. The ability to simulate accurate focusing geometries will greatly enhance the usefulness of Monte Carlo for countless applications, including studying laser tissue interactions in medical applications and light propagation through turbid media.

Bone marrow is both the main hematopoietic and important immune organ. Bone marrow lesions (BMLs) may cause a series of severe complications and even myeloma. The traditional diagnosis of BMLs rely on mostly bone marrow biopsy/ puncture, and sometimes MRI, X-ray, and etc., which are either invasive and dangerous, or ionizing and costly. A diagnosis technology with advantages in noninvasive, safe, real-time continuous detection, and low cost is requested. Here we reported our preliminary exploration of feasibility verification of using near-infrared spectroscopy (NIRS) in clinical diagnosis of BMLs by Monte Carlo simulation study. We simulated and visualized the light propagation in the bone marrow quantitatively with a Monte Carlo simulation software for 3D voxelized media and Visible Chinese Human data set, which faithfully represents human anatomy. The results indicate that bone marrow actually has significant effects on light propagation. According to a sequence of simulation and data analysis, the optimal source-detector separation was suggested to be narrowed down to 2.8–3.2cm, at which separation the spatial sensitivity distribution of NIRS cover the most region of bone marrow with high signal-to-noise ratio. The display of the sources and detectors were optimized as well. This study investigated the light transport in spine addressing to the BMLs detection issue and reported the feasibility of NIRS detection of BMLs noninvasively in theory. The optimized probe design of the coming NIRS-based BMLs detector is also provided.

The analysis of statistical moments of time-resolved (TR) diffuse optical signals can be used to evaluate the absorption and scattering coefficients of turbid media; however, this method requires careful measurement of the instrument response function. We propose an alternative approach that avoids this step by estimating the optical properties from the difference of TR measurements acquired at different source-detector separations. The efficiency of this method was validated using simulated data (from analytical model and Monte-Carlo simulations) and tissue-mimicking phantoms. Results for a homogenous and layered medium showed that the subtraction technique can accurately estimate the optical properties. Specifically, our preliminary results show that the method can estimate the optical properties of a homogeneous medium (simulated using μa = 0.1 mm-1, μs’ = 10 mm-1) with an error less than 10 %. Accurate results were obtained at source-detector separations large enough (5 mm or greater) to resolve differences in the moments. Moreover, we also observed that the subtraction method has improved depth sensitivity compared to the classic method of moments. These results suggests that time-resolved subtraction is a simple but effective means of quantifying optical properties of turbid media, in addition to offering a new approach for obtaining spatially sensitive measurements, although additional studies are required to confirm the latter.

With the introduction of more and more new wavelengths, one of the main problems of medical laser users was centered on the study of laser-tissue interactions with the aim of determining the ideal wavelength for their treatments. The aim of this ex vivo study was to determine, by means of the utilization of a supercontinuum source, the amount of transmitted energy of different wavelengths in different organ samples obtained by Sprague Dawley rats. Supercontinuum light is generated by exploiting high optical non-linearity in a material and it combines the broadband attributes of a lamp with the spatial coherence and high brightness of laser. Even if the single transmission measurement does not allow us to separate out the respective contribution of scattering and absorption, it gives us an evaluation of the wavelengths not interacting with the tissue. In this way, being possible to determine what of the laser wavelengths are not useful or active in the different kinds of tissue, physicians may choose the proper device for his clinical treatments.

In this study, light transmittance of MCF-7 tumor cells from 450 nm to 1100 nm has been measured in their growing medium and evaluated. Transmittance differences have been tried to be put forward in cancer cell line on visible (VIS) and near infrared (NIR) spectrum as well as in between different numbers of cells in medium. An absorption-reflection spectrophotometer was used in the experiments. System has a tungsten light source, optical chopper, a monochromator, sample chamber, silicon detectors, lock-in amplifier and computer. System was controlled by software in order to adjust scan range, scan steps and grating configuration. Cells were grown in medium, and measurements were taken from cells while they were in 5 ml medium. According to our findings, there are significant differences between VIS and NIR regions for the same number of cells. There were found no statistical difference among different numbers of cells. Increasing number of cells has not affected the transmittance. Transmittance of medium is not significantly different from different concentration of cells.

In this paper, diffuse reflectance hyperspectral images of a light beam propagating through a semi-infinite homogeneous layer were simulated by a modified version of the open source Monte Carlo (MC) for multi-layered tissues. Subsequently, the optical properties in terms of absorption and reduced scattering coefficients were extracted from the simulated hyperspectral images by an inverse MC model based on a criterion function that exploits the spatially resolved information of hyperspectral images. The method was validated on real hyperspectral images of turbid phantoms with exactly defined optical properties.

The penetration depth of light plays a crucial role in therapeutic medical applications. In order to design effective medical photonic devices, an in-depth understanding of light’s ability to penetrate tissues (including bone, skin, and fat) is necessary. The amount of light energy absorbed or scattered by tissues affects the intensity of light reaching an intended target in vivo. In this study, we examine the transmittance of light through a variety of cranial tissues for the purpose of determining the efficacy of neuro stimulation using a transcranial laser. Tissue samples collected from a pig were irradiated with a pulsed laser. We first determine the optimal irradiation wavelength of the laser to be 808nm. With varying peak and average power of the laser, we found an inverse and logarithmic relationship between the penetration depth and the intensity of the light. After penetrating the skin and skull of the pig, the light decreases in intensity at a rate of approximately 90.8 (±0.4) percent for every 5 mm of brain tissue penetrated. We also found the correlation between the irradiation time and dosage, using three different lasers (with peak power of 500, 1000, and 1500mW respectively). These data will help deduce what laser power is required to achieve a clinically-realistic model for a given irradiation time. This work is fundamental and the experimental data can be used to supplement existing and future research on the effects of laser light on brain tissue for the design of medical devices.

In this paper, the commonly used inverse Monte Carlo model based on absorption and reduced scattering coefficients is extended by a well-known similarity parameter γ (gamma), which carries additional information on the phase function. Sub-diffuse reflectance measurements at five source-detector separations were used to extract the absorption and reduced scattering coefficients and phase function information encapsulated in γ. The performance of the extended inverse Monte Carlo model was evaluated by simulated and experimental reflectance spectra of turbid phantoms. A three-fold increase in the accuracy of the extended inverse Monte Carlo model that incorporates γ was observed.

Accurate determination of in-vivo light fluence rate is critical for preclinical and clinical studies involving photodynamic therapy (PDT). This study compares the longitudinal light fluence distribution inside biological tissue in the central axis of a 1 cm diameter circular uniform light field for a range of in-vivo tissue optical properties (absorption coefficients (μ_a) between 0.01 and 1 cm^-1 and reduced scattering coefficients (μ_s’) between 2 and 40 cm^-1). This was done using Monte-Carlo simulations for a semi-infinite turbid medium in an air-tissue interface. The end goal is to develop an analytical expression that would fit the results from the Monte Carlo simulation for both the 1 cm diameter circular beam and the broad beam. Each of these parameters is expressed as a function of tissue optical properties. These results can then be compared against the existing expressions in the literature for broad beam for analysis in both accuracy and applicable range. Using the 6-parameter model, the range and accuracy for light transport through biological tissue is improved and may be used in the future as a guide in PDT for light fluence distribution for known tissue optical properties.

Lasers have demonstrated widespread applicability in clinical dermatology as minimally invasive instruments that achieve photogenerated responses within tissue. However, before reaching its target, the incident light must first transmit through the surface layer of tissue, which is interspersed with chromophores (e.g. melanin) that preferentially absorb the light and may also generate negative tissue responses. These optical absorbers decrease the efficacy of the procedures. In order to ensure that the target receives a clinically relevant dose, most procedures simply increase the incident energy; however, this tends to exacerbate the negative complications of melanin absorption. Here, we present an alternative solution aimed at increasing epidermal energy uence while mitigating excess absorption by unintended targets. Our technique involves the combination of a waveguide-based contact transmission modality with simultaneous high-frequency ultrasonic pulsation, which alters the optical properties of the tissue through the agglomeration of dissolved gasses into micro-bubbles within the tissue. Doing so effectively creates optically transparent pathways for the light to transmit unobstructed through the tissue, resulting in an increase in forward scattering and a decrease in absorption. To demonstrate this, Q-switched nanosecond-pulsed laser light at 532nm was delivered into pig skin samples using custom glass waveguides clad in titanium and silver. Light transmission through the tissue was measured with a photodiode and integrating sphere for tissue with and without continuous ultrasonic pulsation at 510 kHz. The combination of these techniques has the potential to improve the efficiency of laser procedures while mitigating negative tissue effects caused by undesirable absorption.

Most analytical methods for describing light propagation in turbid medium exhibit low effectiveness in the near-field of a collimated source. Motivated by the Charge Simulation Method in electromagnetic theory as well as the established discrete source based modeling, we have reported on an improved explicit model, referred to as "Virtual Source" (VS) diffuse approximation (DA), to inherit the mathematical simplicity of the DA while considerably extend its validity in modeling the near-field photon migration in low-albedo medium. In this model, the collimated light in the standard DA is analogously approximated as multiple isotropic point sources (VS) distributed along the incident direction. For performance enhancement, a fitting procedure between the calculated and realistic reflectances is adopted in the nearfield to optimize the VS parameters (intensities and locations). To be practically applicable, an explicit 2VS-DA model is established based on close-form derivations of the VS parameters for the typical ranges of the optical parameters. The proposed VS-DA model is validated by comparing with the Monte Carlo simulations, and further introduced in the image reconstruction of the Laminar Optical Tomography system.

Objective: In this study, the metabolic state of the heart tissue is studied in a rodent model of ischemia and reperfusion (IR) in rats exposed to irradiation injury using a cryofluorescence imaging technique. Mitochondrial metabolic state is evaluated by autofluorescence of mitochondrial metabolic coenzymes NADH and FAD. The redox ratio (NADH/FAD) is used as a biochemical/metabolic marker of oxidative stress, before, during and after IR. Materials and methods: Hearts were extracted from non-irradiated (control) and irradiated rats (Irr) given 15 Gy whole thorax irradiation rats (WTI). After 35 days, before the onset of radiation pneumonitis, these two groups of hearts were subjected to one of three treatments; Time control (TC; hearts perfused for the duration of the protocol without ischemia or IR), 25 minutes ischemia with no reperfusion and 25 minutes ischemia followed by 60 minutes reperfusion (IR). Hearts were removed from the Langendorff perfusion system and immediately snap frozen in liquid N2 to preserve the metabolic state after injury; 3-dimensional (3D) cryo-fluorescent imager was used to obtain in fixed time NADH and FAD fluorescence images and their distribution across the entire ventricles. In this study, a 30-μm axial resolution was used resulting in 550 cross-section images per heart. The 3D images of the redox ratio and their respective histograms were calculated in the six groups of hearts. Results: We compared the mean values of the redox ratio in each group, which demonstrate a reduced mitochondrial redox state in both irradiated and non-irradiated ischemic hearts and an oxidized mitochondrial redox state for both irradiated and non-irradiated ischemia-reperfusion hearts compared to control hearts. For non-irradiated hearts, ischemia and IR injuries resulted respectively in 61% increase and 54% decrease in redox ratio when compared with TC. For irradiated hearts, ischemia and IR injuries resulted respectively in 90% increase and 50% decrease in redox ratio when compared to TC. Conclusion: The cryoimager is able to quantify ischemia and IR injuries. Cryoimaging is a unique 3D imaging tools that provides quantitative measurement of tissue metabolic state. Hearts that underwent irradiation indicates a more oxidized metabolic state in the tissue. This change persists across all three treatments.

In order to investigate patency of heart blood vessels by photosensitization reaction shortly after intravenous injection of talaporfin sodium, we performed in vitro endothelial cell lethality study and in vivo study of heart blood vessel patency in canine one week after photosensitization reaction. Cell lethality of human umbilical vein endothelial cells under different albumin concentrations corresponding with blood and interstice concentrations were employed and their lethality 2 hours after the reaction was measured by WST assay in vitro. Almost all cells survived by 40 J/cm² photosensitization reaction with blood albumin concentration. Laser diffuser made of plastic optical fiber with 70 mm in length was used in vivo. Red diode laser of 664nm wavelength was emitted from this diffuser with 17.1-42.9 mW/cm in 10 minutes. We estimated the fluence rate distribution by a ray-trace simulator using pre-measured optical coefficients of myocardium tissue, μa; 0.12 mm^-1 and μs’; 0.36 mm^-1. Almost all blood vessels were patent in every irradiation conditions in canine heart. Coronary artery and vein up to 1 mm diameter were patent in typical myocardium sample with 25.7 mW/cm. We estimated fluence rate distribution of this sample and found that blood vessels were patent even fluence rate over 40 J/cm². This in vivo study could be explained by the result of in vitro study. We suggest that this blood vessel patency after our particular photosensitization reaction might be because of few photosensitizer uptake in the blood endothelial cells and/or reduced oxidation damage by thick albumin concentration in blood.

Observations of RPE disruption and autofluorescence (AF) photobleaching at light levels below the ANSI photochemical maximum permissible exposure (MPE) (Morgan et al., 2008) indicates a demand to modify future light safety standards to protect the retina from harm. To establish safe light exposures, we measured the visible light action spectrum for RPE disruption in an in vivo monkey model with fluorescence adaptive optics retinal imaging. Using this high resolution imaging modality can provide insight into the consequences of light on a cellular level and allow for longitudinal monitoring of retinal changes. The threshold retinal radiant exposures (RRE) for RPE disruption were determined for 4 wavelengths (460, 488, 544, and 594 nm). The anaesthetized macaque retina was exposed to a uniform 0.5° × 0.5° field of view (FOV). Imaging within a 2° × 2° FOV was performed before, immediately after and at 2 week intervals for 10 weeks. At each wavelength, multiple RREs were tested with 4 repetitions each to determine the threshold for RPE disruption. For qualitative analysis, RPE disruption is defined as any detectable change from the pre exposure condition in the cell mosaic in the exposed region relative to the corresponding mosaic in the immediately surrounding area. We have tested several metrics to evaluate the RPE images obtained before and after exposure. The measured action spectrum for photochemical RPE disruption has a shallower slope than the current ANSI photochemical MPE for the same conditions and suggests that longer wavelength light is more hazardous than other measurements would suggest.

Bioeffects of directed-optical-energy encompass a wide range of applications. One aspect of photochemical interactions involves irradiating a photosensitizer with visible light in order to induce protein unfolding and consequent changes in function. In the past, irradiation of several dye-protein combinations has revealed effects on protein structure. Beta lactoglobulin, human serum albumin (HSA) and tubulin have all been photo-modified with meso-tetrakis(4- sulfonatophenyl)porphyrin (TSPP) bound, but only in the case of tubulin has binding caused a verified loss of biological function (loss of ability to form microtubules) as a result of this light-induced structural change. The current work questions if the photo-induced structural changes that occur to HSA, are sufficient to disable its biological function of binding to osteonectin. The albumin-binding protein, osteonectin, is about half the molecular weight of HSA, so the two proteins and their bound product can be separated and quantified by size exclusion high performance liquid chromatography. TSPP was first bound to HSA and irradiated, photo-modifying the structure of HSA. Then native HSA or photo-modified HSA (both with TSPP bound) were compared, to assess loss in HSA’s innate binding ability as a result of light-induced structure modification.

Mammary epithelial cell (MEC) organoids in 3D culture recapitulate features of breast ducts in vivo. OCT has the ability to monitor the evolution of MEC organoids non-invasively and longitudinally. The anti-cancer drug Doxorubicin (Dox) is able to inhibit proliferation of cancer cells and has been widely used for chemotherapy of breast cancers; while environmental toxins implicated in breast cancer such as estrogen regulates mammary tumor growth and stimulates the proliferation and metastatic potential of breast cancers. Here we propose a quantitative method for measuring motility of breast cells in 3D cultures based upon OCT speckle fluctuation spectroscopy. The metrics of the inverse power-law exponent (α) and fractional modulation amplitude (M) were extracted from speckle fluctuation spectra. These were used to quantify the responses of MEC organoids to Dox, and estrogen. We investigated MEC organoids comprised of two different MEC lines: MCF10DCIS.com exposed to Dox, and MCF7 exposed to estrogen. We found an increase (p<0.001) in α of MEC along time (t=0, 1 hour, 24 hours, 48 hours and 6 days) at each dose of Dox (0, 1 μM and 10 μM), indicating lower fluctuation intensity at higher frequencies. We also observed a decrease (p<0.001) in M for increasing time. However, both α and M of MCF7 treated with estrogen (0, 1 nM and 10 nM) exhibited the opposite trend along time. This novel technology provides rapid and non-invasive measurements of the effects of toxicants on MEC motility for understanding breast cancer development and assessing anti-cancer drugs.

Photothermal damage rate processes in biological tissues are usually characterized by a kinetics approach. This stems from experimental data that show how the transformation of a specified biological property of cells or biomolecule (plating efficiency for viability, change in birefringence, tensile strength, etc.) is dependent upon both time and temperature. However, kinetic methods require determination of kinetic rate constants and knowledge of substrate or product concentrations during the reaction. To better understand photothermal damage processes we have identified temperature histories of cultured retinal cells receiving minimum lethal thermal doses for a variety of laser and culture parameters. These “threshold” temperature histories are of interest because they inherently contain information regarding the fundamental thermal dose requirements for damage in individual cells. We introduce the notion of time-integrated temperature (Tint) as an accumulated thermal dose (ATD) with units of °C s. Damaging photothermal exposure raises the rate of ATD accumulation from that of the ambient (e.g. 37 °C) to one that correlates with cell death (e.g. 52 °C). The degree of rapid increase in ATD (ΔATD) during photothermal exposure depends strongly on the laser exposure duration and the ambient temperature.

Deep vein thrombosis (DVT) always induced venous thrombosis. Most cases of venous thrombosis were induced by deep vein thrombosis (DVT), with high incidence rate of >60% in >60 years old people. Near-infrared spectroscopy (NIRS) were reported recently to be an intriguing and potential technique in detecting DVT in clinics. However, the photon transport is still unclear, which is crucial for the image reconstruction of the updated development called as NIRS-based DVT imager. Here we employed the Monte Carlo simulation software for 3D voxelized media (MCVM) and the Visible Chinese Human (VCH) model, which segmentation is finest in the world, to simulate multi-channel photon migration in calf muscle. And the image reconstruction of DVT hemodynamic distribution was achieved. This study, for the first time, provides the most realistic 3-D multichannel photon migration for NIRS study on DVT, and explored the image reconstruction for furtherly developing a NIRS-based DVT imager.

Oblique light stimulation evoked transient retinal phototropism (TRP) has been recently detected in frog and mouse retinas. High resolution microscopy of freshly isolated retinas indicated that the TRP is predominated by rod photoreceptors. Comparative confocal microscopy and optical coherence tomography (OCT) revealed that the TRP predominantly occurred from the photoreceptor outer segment (OS). However, biophysical mechanism of rod OS change is still unknown. In this study, frog retinal slices, which open a cross section of retinal photoreceptor and other functional layers, were used to test the effect of light stimulation on rod OS. Near infrared light microscopy was employed to monitor photoreceptor changes in retinal slices stimulated by a rectangular-shaped visible light flash. Rapid rod OS length change was observed after the stimulation delivery. The magnitude and direction of the rod OS change varied with the position of the rods within the stimulated area. In the center of stimulated region the length of the rod OS shrunk, while in the peripheral region the rod OS tip swung towards center region in the plane perpendicular to the incident stimulus light. Our experimental result and theoretical analysis suggest that the observed TRP may reflect unbalanced disc-shape change due to localized pigment bleaching. Further investigation is required to understand biochemical mechanism of the observed rod OS kinetics. Better study of the TRP may provide a noninvasive biomarker to enable early detection of age-related macular degeneration (AMD) and other diseases that are known to produce retinal photoreceptor dysfunctions.

Medical laser applications are promoted as safe, effective treatments for a multiplicity of concerns, ranging from hyperthermal skin rejuvenation to subcutaneous tumor ablation. Chromophore and structural protein concentration and distribution within a patient’s tissue vary from patient to patient and dictate the interaction of incident radiative energy of a specific wavelength with the target tissue. Laser parameters must be matched to tissue optical and thermal properties in order to achieve the desired therapeutic results without inducing unnecessary tissue damage, although accurate tissue optical properties are not always measured prior to and during laser therapies. A weighted variable step size Monte Carlo simulation of laser irradiation of skin tissue was used to determine the effects of variations in absorption (μ_a) and scattering coefficients (μs) and the degree of anisotropy (g) on the radiant energy transport per mm² in response to steady-state photon propagation. The three parameters were varied in a factorial experimental design for the ranges of 0.25/mm ≤ μ_a ≤ 2.0/mm, 30.0/mm ≤ μ_s ≤ 140.0/mm, and 0.65 ≤ g ≤ 0.99 in order to isolate their impacts on the overall fluence distribution. Box plots of the resulting fluence profiles were created and compared to identify ranges in which optical property variance could be considered to significantly impact the spatial variance of fluence within the simulation volume. Results indicated that accurate prediction of the fluence profiles that will be achieved by any given medical laser treatment is unlikely without pre-treatment assessment of the tissue optical properties of individual patients.

The experimental results presented in this study are the early studies of germination on the example of Picea abies and were aimed at testing the germination of seeds and the development of morphology, caused a therapeutic effect on the laser radiation field in the early stages of development under the action of ultraviolet and red light in the spectral range of 405 nm and 640 nm. A set of seeds irradiated at various energy doses within the same time. The experimental results analyzed in parallel with control group. In all analyzed seeds were studied the germination and growth of seedlings. The results showed that the percentage of germination higher than control group Samanids all of the recurrence options.

We have studied several sensor concepts for biomedical applications operating in the millimeter wave and terahertz range. On one hand, rectangular waveguide structure were designed and extended with microfluidic channels. In this way a simple analysis of aqueous solutions at various waveguide bands is possible. In our case, we focused on the frequency range between 75 GHz and 110 GHz. On the other hand, planar sensor structures for aqueous solutions have been developed based on coplanar waveguides. With these planar sensors it is possible to concentrate the interaction volume on small sensor areas, which achieve a local exposure of the radiation to the sample. When equipping the sensor with microfluidic structures the sample volume could be reduced significantly and enabled a localized interaction with the sensor areas. The sensors are designed to exhibit a broadband behavior up to 300 GHz. Narrow-band operation can also be achieved for potentially increased sensitivity by using resonant structures. Several tests with Glucose dissolved in water show promising results for the distinction of different glucose levels at millimeter wave frequencies. The planar structures can also be used for the exposure of biological cells or cell model systems like liposomes with electromagnetic radiation. Several studies are planned to distinguish on one hand the influence of millimeter wave exposure on biological systems and also to have a spectroscopic method which enables the analysis of cell processes, like membrane transport processes, with millimeter wave and terahertz frequencies by focusing the electric field directly on the analyzing sample.

This work aims to enact a quick and reasonable estimation of breast cancer margin thickness using time of flight analysis of embedded breast cancer tissue. A pulsed terahertz system is used to obtain reflection imaging scans from breast cancer tumors that are formalin-fixed and embedded in paraffin blocks. Time of flight analysis is then used to compare the reflection patterns seen within the block to pathology sections and paraffin-embedded sections that are taken throughout the depth of the tumor in order to estimate the three-dimensional boundaries of the tumor.

This work seeks to obtain the properties of paraffin-embedded breast cancer tumor tissues using transmission imaging and spectroscopy. Formalin-fixed and paraffin-embedded breast tumors are first sectioned into slices of 20 μm and 30 μm and placed between two tsurupica slides. The slides are then scanned in a pulsed terahertz system using transmission imaging. The tissue regions in adjacent pathology section are compared to the transmission imaging scan in order to define a region of points over which to average the electrical properties results from the scan.

Rapid development in nanomaterial synthesis and functionalization has led to advanced studies in actuation and manipulation of cellular functions for biomedical applications. Often these actuation techniques employ externally applied magnetic fields to manipulate magnetic nanomaterials inside cell bodies in order to drive or trigger desired effects. While cellular interactions with low-frequency magnetic fields and nanoparticles have been extensively studied, the fundamental mechanisms behind these interactions remain poorly understood. Additionally, modern investigations on these concurrent exposure conditions have been limited in scope, and difficult to reproduce. This study presents an easily reproducible method of investigating the biological impact of concurrent magnetic field and nanoparticle exposure conditions using an in-vitro CHO-K1 cell line model, with the purpose of establishing grounds for in-depth fundamental studies of the mechanisms driving cellular-level interactions. Cells were cultured under various nanoparticle and magnetic field exposure conditions from 0 to 500 μg/ml nanoparticle concentrations, and DC, 50 Hz, or 100 Hz magnetic fields with 2.0 mT flux density. Cells were then observed by confocal fluorescence microscopy, and subject to biological assays to determine the effects of concurrent extreme-low frequency magnetic field and nanoparticle exposures on cellnanoparticle interactions, such as particle uptake and cell viability by MTT assay. Current results indicate little to no variation in effect on cell cultures based on magnetic field parameters alone; however, it is clear that deleterious synergistic effects of concurrent exposure conditions exist based on a significant decrease in cell viability when exposed to high concentrations of nanoparticles and concurrent magnetic field.