Showing papers in "Biomedical Optics Express in 2013"

High-resolution, lensless endoscope based on digital scanning through a multimode optical fiber

Abstract: We propose and experimentally demonstrate an ultra-thin rigid endoscope (450 μm diameter) based on a passive multimode optical fiber. We use digital phase conjugation to overcome the modal scrambling of the fiber to tightly focus and scan the laser light at its distal end. By exploiting the maximum number of modes available, sub-micron resolution, high quality fluorescence images of neuronal cells were acquired. The imaging system is evaluated in terms of fluorescence collection efficiency, resolution and field of view. The small diameter of the proposed endoscope, along with its high quality images offer an opportunity for minimally invasive medical endoscopic imaging and diagnosis based on cellular phenotype via direct tissue penetration.

Retinal layer segmentation of macular OCT images using boundary classification

Abstract: Optical coherence tomography (OCT) has proven to be an essential imaging modality for ophthalmology and is proving to be very important in neurology OCT enables high resolution imaging of the retina, both at the optic nerve head and the macula Macular retinal layer thicknesses provide useful diagnostic information and have been shown to correlate well with measures of disease severity in several diseases Since manual segmentation of these layers is time consuming and prone to bias, automatic segmentation methods are critical for full utilization of this technology In this work, we build a random forest classifier to segment eight retinal layers in macular cube images acquired by OCT The random forest classifier learns the boundary pixels between layers, producing an accurate probability map for each boundary, which is then processed to finalize the boundaries Using this algorithm, we can accurately segment the entire retina contained in the macular cube to an accuracy of at least 43 microns for any of the nine boundaries Experiments were carried out on both healthy and multiple sclerosis subjects, with no difference in the accuracy of our algorithm found between the groups

Autoregressive model based algorithm for correcting motion and serially correlated errors in fNIRS

Abstract: Systemic physiology and motion-induced artifacts represent two major sources of confounding noise in functional near infrared spectroscopy (fNIRS) imaging that can reduce the performance of analyses and inflate false positive rates (i.e., type I errors) of detecting evoked hemodynamic responses. In this work, we demonstrated a general algorithm for solving the general linear model (GLM) for both deconvolution (finite impulse response) and canonical regression models based on designing optimal pre-whitening filters using autoregressive models and employing iteratively reweighted least squares. We evaluated the performance of the new method by performing receiver operating characteristic (ROC) analyses using synthetic data, in which serial correlations, motion artifacts, and evoked responses were controlled via simulations, as well as using experimental data from children (3–5 years old) as a source baseline physiological noise and motion artifacts. The new method outperformed ordinary least squares (OLS) with no motion correction, wavelet based motion correction, or spline interpolation based motion correction in the presence of physiological and motion related noise. In the experimental data, false positive rates were as high as 37% when the estimated p-value was 0.05 for the OLS methods. The false positive rate was reduced to 5–9% with the proposed method. Overall, the method improves control of type I errors and increases performance when motion artifacts are present.

Blood flow velocity quantification using split-spectrum amplitude-decorrelation angiography with optical coherence tomography

Abstract: The split-spectrum amplitude-decorrelation angiography (SSADA) algorithm was recently developed as a method for imaging blood flow in the human retina without the use of phase information. In order to enable absolute blood velocity quantification, in vitro phantom experiments are performed to correlate the SSADA signal at multiple time scales with various preset velocities. A linear model relating SSADA measurements to absolute flow velocities is derived using the phantom data. The operating range for the linear model is discussed along with its implication for velocity quantification with SSADA in a clinical setting.

Multi-MHz retinal OCT

Abstract: We analyze the benefits and problems of in vivo optical coherence tomography (OCT) imaging of the human retina at A-scan rates in excess of 1 MHz, using a 1050 nm Fourier-domain mode-locked (FDML) laser. Different scanning strategies enabled by MHz OCT line rates are investigated, and a simple multi-volume data processing approach is presented. In-vivo OCT of the human ocular fundus is performed at different axial scan rates of up to 6.7 MHz. High quality non-mydriatic retinal imaging over an ultra-wide field is achieved by a combination of several key improvements compared to previous setups. For the FDML laser, long coherence lengths and 72 nm wavelength tuning range are achieved using a chirped fiber Bragg grating in a laser cavity at 419.1 kHz fundamental tuning rate. Very large data sets can be acquired with sustained data transfer from the data acquisition card to host computer memory, enabling high-quality averaging of many frames and of multiple aligned data sets. Three imaging modes are investigated: Alignment and averaging of 24 data sets at 1.68 MHz axial line rate, ultra-dense transverse sampling at 3.35 MHz line rate, and dual-beam imaging with two laser spots on the retina at an effective line rate of 6.7 MHz.

Re-scan confocal microscopy: scanning twice for better resolution

Abstract: We present a new super-resolution technique, Re-scan Confocal Microscopy (RCM), based on standard confocal microscopy extended with an optical (re-scanning) unit that projects the image directly on a CCD-camera. This new microscope has improved lateral resolution and strongly improved sensitivity while maintaining the sectioning capability of a standard confocal microscope. This simple technology is typically useful for biological applications where the combination high-resolution and high-sensitivity is required.

Automated non-rigid registration and mosaicing for robust imaging of distinct retinal capillary beds using speckle variance optical coherence tomography

Abstract: Variance processing methods in Fourier domain optical coherence tomography (FD-OCT) have enabled depth-resolved visualization of the capillary beds in the retina due to the development of imaging systems capable of acquiring A-scan data in the 100 kHz regime. However, acquisition of volumetric variance data sets still requires several seconds of acquisition time, even with high speed systems. Movement of the subject during this time span is sufficient to corrupt visualization of the vasculature. We demonstrate a method to eliminate motion artifacts in speckle variance FD-OCT images of the retinal vasculature by creating a composite image from multiple volumes of data acquired sequentially. Slight changes in the orientation of the subject’s eye relative to the optical system between acquired volumes may result in non-rigid warping of the image. Thus, we use a B-spline based free form deformation method to automatically register variance images from multiple volumes to obtain a motion-free composite image of the retinal vessels. We extend this technique to automatically mosaic individual vascular images into a widefield image of the retinal vasculature.

Terahertz imaging of excised oral cancer at frozen temperature

Abstract: The feasibility of terahertz (THz) imaging at frozen temperature for the clinical application of oral cancer detection was investigated by analyzing seven oral tissues resected from four patients. The size, shape, and internal position of the oral cancers were mapped by THz radiation in the frequency range of 0.2–1.2 THz at −20 °C and 20 °C, and compared with those identified in the histological examination. THz imaging of frozen tissue was found to offer greater sensitivity in distinguishing cancerous areas from surrounding tissue and a larger THz-frequency spectral difference between the oral cancer and normal mucosa than room-temperature THz imaging. A cancerous tumor hidden inside tissue was also detected using this method by observing the THz temporal domain waveform. The histological analysis showed that these findings resulted from cell structure deformations involving the invasion of oral tumor and neoplastic transformations of mucous cells. Therefore, a cytological approach using THz radiation at a frozen temperature might be applied to detect oral cancer.

Intense THz pulses cause H2AX phosphorylation and activate DNA damage response in human skin tissue

Abstract: Recent emergence and growing use of terahertz (THz) radiation for medical imaging and public security screening raise questions on reasonable levels of exposure and health consequences of this form of electromagnetic radiation. In particular, picosecond-duration THz pulses have shown promise for novel diagnostic imaging techniques. However, the effects of THz pulses on human cells and tissues thus far remain largely unknown. We report on the investigation of the biological effects of pulsed THz radiation on artificial human skin tissues. We observe that exposure to intense THz pulses for ten minutes leads to a significant induction of H2AX phosphorylation, indicating that THz pulse irradiation may cause DNA damage in exposed skin tissue. At the same time, we find a THz-pulse-induced increase in the levels of several proteins responsible for cell-cycle regulation and tumor suppression, suggesting that DNA damage repair mechanisms are quickly activated. Furthermore, we find that the cellular response to pulsed THz radiation is significantly different from that induced by exposure to UVA (400 nm).

Ultrahigh speed endoscopic optical coherence tomography using micromotor imaging catheter and VCSEL technology

Abstract: We developed a micromotor based miniature catheter with an outer diameter of 3.2 mm for ultrahigh speed endoscopic swept source optical coherence tomography (OCT) using a vertical cavity surface-emitting laser (VCSEL) at a 1 MHz axial scan rate. The micromotor can rotate a micro-prism at several hundred frames per second with less than 5 V drive voltage to provide fast and stable scanning, which is not sensitive to the bending of the catheter. The side-viewing probe can be pulled back to acquire a three-dimensional (3D) data set covering a large area on the specimen. The VCSEL provides a high axial scan rate to support dense sampling under high frame rate operation. Using a high speed data acquisition system, in vivo 3D-OCT imaging in the rabbit GI tract and ex vivo imaging of a human colon specimen with 8 μm axial resolution, 8 μm lateral resolution and 1.2 mm depth range in tissue at a frame rate of 400 fps was demonstrated.

In vivo dark-field imaging of the retinal pigment epithelium cell mosaic.

Abstract: Non-invasive reflectance imaging of the human RPE cell mosaic is demonstrated using a modified confocal adaptive optics scanning light ophthalmoscope (AOSLO). The confocal circular aperture in front of the imaging detector was replaced with a combination of a circular aperture 4 to 16 Airy disks in diameter and an opaque filament, 1 or 3 Airy disks thick. This arrangement reveals the RPE cell mosaic by dramatically attenuating the light backscattered by the photoreceptors. The RPE cell mosaic was visualized in all 7 recruited subjects at multiple retinal locations with varying degrees of contrast and cross-talk from the photoreceptors. Various experimental settings were explored for improving the visualization of the RPE cell boundaries including: pinhole diameter, filament thickness, illumination and imaging pupil apodization, unmatched imaging and illumination focus, wavelength and polarization. None of these offered an obvious path for enhancing image contrast. The demonstrated implementation of dark-field AOSLO imaging using 790 nm light requires low light exposures relative to light safety standards and it is more comfortable for the subject than the traditional autofluorescence RPE imaging with visible light. Both these factors make RPE dark-field imaging appealing for studying mechanisms of eye disease, as well as a clinical tool for screening and monitoring disease progression.

Real-time eye motion correction in phase-resolved OCT angiography with tracking SLO

Abstract: In phase-resolved OCT angiography blood flow is detected from phase changes in between A-scans that are obtained from the same location. In ophthalmology, this technique is vulnerable to eye motion. We address this problem by combining inter-B-scan phase-resolved OCT angiography with real-time eye tracking. A tracking scanning laser ophthalmoscope (TSLO) at 840 nm provided eye tracking functionality and was combined with a phase-stabilized optical frequency domain imaging (OFDI) system at 1040 nm. Real-time eye tracking corrected eye drift and prevented discontinuity artifacts from (micro)saccadic eye motion in OCT angiograms. This improved the OCT spot stability on the retina and consequently reduced the phase-noise, thereby enabling the detection of slower blood flows by extending the inter-B-scan time interval. In addition, eye tracking enabled the easy compounding of multiple data sets from the fovea of a healthy volunteer to create high-quality eye motion artifact-free angiograms. High-quality images are presented of two distinct layers of vasculature in the retina and the dense vasculature of the choroid. Additionally we present, for the first time, a phase-resolved OCT angiogram of the mesh-like network of the choriocapillaris containing typical pore openings.

Automated tissue characterization of in vivo atherosclerotic plaques by intravascular optical coherence tomography images

Abstract: Intravascular optical coherence tomography (IVOCT) is rapidly becoming the method of choice for the in vivo investigation of coronary artery disease. While IVOCT visualizes atherosclerotic plaques with a resolution <20µm, image analysis in terms of tissue composition is currently performed by a time-consuming manual procedure based on the qualitative interpretation of image features. We illustrate an algorithm for the automated and systematic characterization of IVOCT atherosclerotic tissue. The proposed method consists in a supervised classification of image pixels according to textural features combined with the estimated value of the optical attenuation coefficient. IVOCT images of 64 plaques, from 49 in vivo IVOCT data sets, constituted the algorithm’s training and testing data sets. Validation was obtained by comparing automated analysis results to the manual assessment of atherosclerotic plaques. An overall pixel-wise accuracy of 81.5% with a classification feasibility of 76.5% and per-class accuracy of 89.5%, 72.1% and 79.5% for fibrotic, calcified and lipid-rich tissue respectively, was found. Moreover, measured optical properties were in agreement with previous results reported in literature. As such, an algorithm for automated tissue characterization was developed and validated using in vivo human data, suggesting that it can be applied to clinical IVOCT data. This might be an important step towards the integration of IVOCT in cardiovascular research and routine clinical practice.

Automatic segmentation of choroidal thickness in optical coherence tomography

Abstract: The assessment of choroidal thickness from optical coherence tomography (OCT) images of the human choroid is an important clinical and research task, since it provides valuable information regarding the eye’s normal anatomy and physiology, and changes associated with various eye diseases and the development of refractive error. Due to the time consuming and subjective nature of manual image analysis, there is a need for the development of reliable objective automated methods of image segmentation to derive choroidal thickness measures. However, the detection of the two boundaries which delineate the choroid is a complicated and challenging task, in particular the detection of the outer choroidal boundary, due to a number of issues including: (i) the vascular ocular tissue is non-uniform and rich in non-homogeneous features, and (ii) the boundary can have a low contrast. In this paper, an automatic segmentation technique based on graph-search theory is presented to segment the inner choroidal boundary (ICB) and the outer choroidal boundary (OCB) to obtain the choroid thickness profile from OCT images. Before the segmentation, the B-scan is pre-processed to enhance the two boundaries of interest and to minimize the artifacts produced by surrounding features. The algorithm to detect the ICB is based on a simple edge filter and a directional weighted map penalty, while the algorithm to detect the OCB is based on OCT image enhancement and a dual brightness probability gradient. The method was tested on a large data set of images from a pediatric (1083 B-scans) and an adult (90 B-scans) population, which were previously manually segmented by an experienced observer. The results demonstrate the proposed method provides robust detection of the boundaries of interest and is a useful tool to extract clinical data.

3D static elastography at the micrometer scale using Full Field OCT.

Abstract: Full-Field OCT (FF-OCT) is able to image biological tissues in 3D with micrometer resolution. In this study we add elastographic contrast to the FF-OCT modality. By combining FF-OCT with elastography, we create a virtual palpation map at the micrometer scale. We present here a proof of concept on multi-layer phantoms and preliminary results on ex vivo biological samples such as porcine cornea, human breast tissues and rat heart. The 3D digital volume correlation that is used in connection with the 3D stack of images allows to access to the full 3D strain tensor and to reveal stiffness anisotropy.

Simple buffers for 3D STORM microscopy

Abstract: 3D STORM is one of the leading methods for super-resolution imaging, with resolution down to 10 nm in the lateral direction, and 30–50 nm in the axial direction. However, there is one important requirement to perform this type of imaging: making dye molecules blink. This usually relies on the utilization of complex buffers, containing different chemicals and sensitive enzymatic systems, limiting the reproducibility of the method. We report here that the commercial mounting medium Vectashield can be used for STORM of Alexa-647, and yields images comparable or superior to those obtained with more complex buffers, especially for 3D imaging. We expect that this advance will promote the versatile utilization of 3D STORM by removing one of its entry barriers, as well as provide a more reproducible way to compare optical setups and data processing algorithms.

Single snapshot imaging of optical properties.

Abstract: A novel acquisition and processing method that enables single snapshot wide field imaging of optical properties in the Spatial Frequency Domain (SFD) is described. This method makes use of a Fourier transform performed on a single image and processing in the frequency space to extract two spatial frequency images at once. The performance of the method is compared to the standard six image SFD acquisition method, assessed on tissue mimicking phantoms and in vivo. Overall both methods perform similarly in extracting optical properties.

Low-cost laser speckle contrast imaging of blood flow using a webcam.

Abstract: Laser speckle contrast imaging has become a widely used tool for dynamic imaging of blood flow, both in animal models and in the clinic. Typically, laser speckle contrast imaging is performed using scientific-grade instrumentation. However, due to recent advances in camera technology, these expensive components may not be necessary to produce accurate images. In this paper, we demonstrate that a consumer-grade webcam can be used to visualize changes in flow, both in a microfluidic flow phantom and in vivo in a mouse model. A two-camera setup was used to simultaneously image with a high performance monochrome CCD camera and the webcam for direct comparison. The webcam was also tested with inexpensive aspheric lenses and a laser pointer for a complete low-cost, compact setup ($90, 5.6 cm length, 25 g). The CCD and webcam showed excellent agreement with the two-camera setup, and the inexpensive setup was used to image dynamic blood flow changes before and after a targeted cerebral occlusion.

In vivo imaging of cerebral energy metabolism with two-photon fluorescence lifetime microscopy of NADH.

Abstract: Minimally invasive, specific measurement of cellular energy metabolism is crucial for understanding cerebral pathophysiology. Here, we present high-resolution, in vivo observations of autofluorescence lifetime as a biomarker of cerebral energy metabolism in exposed rat cortices. We describe a customized two-photon imaging system with time correlated single photon counting detection and specialized software for modeling multiple-component fits of fluorescence decay and monitoring their transient behaviors. In vivo cerebral NADH fluorescence suggests the presence of four distinct components, which respond differently to brief periods of anoxia and likely indicate different enzymatic formulations. Individual components show potential as indicators of specific molecular pathways involved in oxidative metabolism.

In vivo imaging of retinal pigment epithelium cells in age related macular degeneration

Abstract: Morgan and colleagues demonstrated that the RPE cell mosaic can be resolved in the living human eye non-invasively by imaging the short-wavelength autofluorescence using an adaptive optics (AO) ophthalmoscope. This method, based on the assumption that all subjects have the same longitudinal chromatic aberration (LCA) correction, has proved difficult to use in diseased eyes, and in particular those affected by age-related macular degeneration (AMD). In this work, we improve Morgan’s method by accounting for chromatic aberration variations by optimizing the confocal aperture axial and transverse placement through an automated iterative maximization of image intensity. The increase in image intensity after algorithmic aperture placement varied depending upon patient and aperture position prior to optimization but increases as large as a factor of 10 were observed. When using a confocal aperture of 3.4 Airy disks in diameter, images were obtained using retinal radiant exposures of less than 2.44 J/cm2, which is ~22 times below the current ANSI maximum permissible exposure. RPE cell morphologies that were strikingly similar to those seen in postmortem histological studies were observed in AMD eyes, even in areas where the pattern of fluorescence appeared normal in commercial fundus autofluorescence (FAF) images. This new method can be used to study RPE morphology in AMD and other diseases, providing a powerful tool for understanding disease pathogenesis and progression, and offering a new means to assess the efficacy of treatments designed to restore RPE health.

Automatic segmentation of the choroid in enhanced depth imaging optical coherence tomography images

Abstract: Enhanced Depth Imaging (EDI) optical coherence tomography (OCT) provides high-definition cross-sectional images of the choroid in vivo, and hence is used in many clinical studies However, the quantification of the choroid depends on the manual labelings of two boundaries, Bruch's membrane and the choroidal-scleral interface This labeling process is tedious and subjective of inter-observer differences, hence, automatic segmentation of the choroid layer is highly desirable In this paper, we present a fast and accurate algorithm that could segment the choroid automatically Bruch's membrane is detected by searching the pixel with the biggest gradient value above the retinal pigment epithelium (RPE) and the choroidal-scleral interface is delineated by finding the shortest path of the graph formed by valley pixels using Dijkstra's algorithm The experiments comparing automatic segmentation results with the manual labelings are conducted on 45 EDI-OCT images and the average of Dice's Coefficient is 905%, which shows good consistency of the algorithm with the manual labelings The processing time for each image is about 125 seconds

Label-free high-throughput cell screening in flow.

Abstract: Flow cytometry is a powerful tool for cell counting and biomarker detection in biotechnology and medicine especially with regards to blood analysis. Standard flow cytometers perform cell type classification both by estimating size and granularity of cells using forward- and side-scattered light signals and through the collection of emission spectra of fluorescently-labeled cells. However, cell surface labeling as a means of marking cells is often undesirable as many reagents negatively impact cellular viability or provide activating/inhibitory signals, which can alter the behavior of the desired cellular subtypes for downstream applications or analysis. To eliminate the need for labeling, we introduce a label-free imaging-based flow cytometer that measures size and cell protein concentration simultaneously either as a stand-alone instrument or as an add-on to conventional flow cytometers. Cell protein concentration adds a parameter to cell classification, which improves the specificity and sensitivity of flow cytometers without the requirement of cell labeling. This system uses coherent dispersive Fourier transform to perform phase imaging at flow speeds as high as a few meters per second.

Short-lag spatial coherence beamforming of photoacoustic images for enhanced visualization of prostate brachytherapy seeds

Abstract: Prostate brachytherapy, administered by implanting tiny radioactive seeds to treat prostate cancer, currently relies on transrectal ultrasound imaging for intraoperative visualization of the metallic seeds. Photoacoustic (PA) imaging has been suggested as a feasible alternative to ultrasound imaging due to its superior sensitivity to metal surrounded by tissue. However, PA images suffer from poor contrast when seeds are distant from the light source. We propose a transperineal light delivery method and investigate the application of a short-lag spatial coherence (SLSC) beamformer to enhance low-contrast photoacoustic signals that are distant from this type of light source. Performance is compared to a conventional delay-and-sum beamformer. A pure gelatin phantom was implanted with black ink-coated brachytherapy seeds and the mean contrast was improved by 3–25 dB with the SLSC beamformer for fiber-seed distances ranging 0.6–6.3 cm, when approximately 10% of the receive aperture elements were included in the short-lag sum. For fiber-seed distances greater than 3–4 cm, the mean contrast-to-noise ratio (CNR) was approximately doubled with the SLSC beamformer, while mean signal-to-noise ratios (SNR) were mostly similar with both beamformers. Lateral resolution was decreased by 2 mm, but improved with larger short-lag values at the expense of poorer CNR and SNR. Similar contrast and CNR improvements were achieved with an uncoated brachytherapy seed implanted in ex vivo tissue. Results indicate that the SLSC beamformer has potential to enhance the visualization of prostate brachytherapy seeds that are distant from the light source.

Improvement of axial resolution and contrast in temporally focused widefield two-photon microscopy with structured light illumination

Abstract: Although temporally focused wide-field two-photon microscopy (TFM) can perform depth resolved wide field imaging, it cannot avoid the image degradation due to scattering of excitation and emission photons when imaging in a turbid medium. Further, its axial resolution is inferior to standard point-scanning two-photon microscopy. We implemented a structured light illumination for TFM and have shown that it can effectively reject the out-of-focus scattered emission photons improving image contrast. Further, the depth resolution of the improved system is dictated by the spatial frequency of the structure light with the potential of attaining depth resolution better than point-scanning two-photon microscopy.

Imaging myocardial fiber orientation using polarization sensitive optical coherence tomography.

Abstract: Knowledge of myocardial fiber architecture is essential towards understanding heart functions. We demonstrated in this study a method to map cardiac muscle structure using the local optical axis obtained from polarization-sensitive optical coherence tomography (PSOCT). An algorithm was developed to extract the true local depth-resolved optical axis, retardance, and diattenuation from conventional round-trip results obtained in a Jones matrix-based PSOCT system. This method was applied to image the myocardial fiber orientation in a bovine heart muscle sample.

Multi-modal approach using Raman spectroscopy and optical coherence tomography for the discrimination of colonic adenocarcinoma from normal colon

Abstract: We report a multimodal optical approach using both Raman spectroscopy and optical coherence tomography (OCT) in tandem to discriminate between colonic adenocarcinoma and normal colon. Although both of these non-invasive techniques are capable of discriminating between normal and tumour tissues, they are unable individually to provide both the high specificity and high sensitivity required for disease diagnosis. We combine the chemical information derived from Raman spectroscopy with the texture parameters extracted from OCT images. The sensitivity obtained using Raman spectroscopy and OCT individually was 89% and 78% respectively and the specificity was 77% and 74% respectively. Combining the information derived using the two techniques increased both sensitivity and specificity to 94% demonstrating that combining complementary optical information enhances diagnostic accuracy. These data demonstrate that multimodal optical analysis has the potential to achieve accurate non-invasive cancer diagnosis.

Chiral imaging of collagen by second-harmonic generation circular dichroism

Abstract: We provide evidence that the chirality of collagen can give rise to strong second-harmonic generation circular dichroism (SHG-CD) responses in nonlinear microscopy. Although chirality is an intrinsic structural property of collagen, most of the previous studies ignore that property. We demonstrate chiral imaging of individual collagen fibers by using a laser scanning microscope and type-I collagen from pig ligaments. 100% contrast level of SHG-CD is achieved with sub-micrometer spatial resolution. As a new contrast mechanism for imaging chiral structures in bio-tissues, this technique provides information about collagen morphology and three-dimensional orientation of collagen molecules.

Flexible delivery of Er:YAG radiation at 2.94 µm with negative curvature silica glass fibers: a new solution for minimally invasive surgical procedures.

Abstract: We present the delivery of high energy microsecond pulses through a hollow-core negative-curvature fiber at 2.94 µm. The energy densities delivered far exceed those required for biological tissue manipulation and are of the order of 2300 J/cm2. Tissue ablation was demonstrated on hard and soft tissue in dry and aqueous conditions with no detrimental effects to the fiber or catastrophic damage to the end facets. The energy is guided in a well confined single mode allowing for a small and controllable focused spot delivered flexibly to the point of operation. Hence, a mechanically and chemically robust alternative to the existing Er:YAG delivery systems is proposed which paves the way for new routes for minimally invasive surgical laser procedures.

Gold nanoparticle targeted photoacoustic cavitation for potential deep tissue imaging and therapy.

Abstract: The laser generation of vapor bubbles around plasmonic nanoparticles can be enhanced through the application of an ultrasound field; a technique referred to as photoacoustic cavitation. The combination of light and ultrasound allows for bubble formation at lower laser fluence and peak negative ultrasound pressure than can be achieved using either modality alone. The growth and collapse of these bubbles leads to local mechanical disruption and acoustic emission, and can potentially be used to induce and monitor tissue therapy. Photoacoustic cavitation is investigated for a broad range of ultrasound pressures and nanoparticle concentrations for gold nanorods and nanospheres. The cavitation threshold fluences for both nanoparticle types are found to drastically reduce in the presence of an ultrasound field. The results indicate that photoacoustic cavitation can potentially be produced at depth in biological tissue without exceeding the safety limits for ultrasound or laser radiation at the tissue surface.

Two-photon excitation in scattering media by spatiotemporally shaped beams and their application in optogenetic stimulation

Abstract: The use of wavefront shaping to generate extended optical excitation patterns which are confined to a predetermined volume has become commonplace on various microscopy applications. For multiphoton excitation, three-dimensional confinement can be achieved by combining the technique of temporal focusing of ultra-short pulses with different approaches for lateral light shaping, including computer generated holography or generalized phase contrast. Here we present a theoretical and experimental study on the effect of scattering on the propagation of holographic beams with and without temporal focusing. Results from fixed and acute cortical slices show that temporally focused spatial patterns are extremely robust against the effects of scattering and this permits their three-dimensionally confined excitation for depths more than 500 µm. Finally we prove the efficiency of using temporally focused holographic beams in two-photon stimulation of neurons expressing the red-shifted optogenetic channel C1V1.