828 kHz retinal imaging with an 840†nm Fourier domain mode locked laser.

This paper presents a Fourier domain mode locked (FDML) laser centered around 840 nm. It features a bidirectional sweep repetition rate of 828 kHz and a spectral bandwidth of 40 nm. An axial resolution of ∼9.9 µm in water and a 1.4 cm sensitivity roll-off are achieved. Utilizing a complex master-slave (CMS) recalibration method and due to a sufficiently high sensitivity of 84.6 dB, retinal layers of the human eye in-vivo can be resolved during optical coherence tomography (OCT) examination. The developed FDML laser enables acquisition rates of 3D-volumes with a size of 200 × 100 × 256 voxels in under 100 milliseconds. Detailed information on the FDML implementation, its challenging design tasks, and OCT images obtained with the laser are presented in this paper.

Effect of an Anaerobic Fermentation Process on 3D-Printed PLA Materials of a Biogas-Generating Reactor.

3D-printed materials are present in numerous applications, from medicine to engineering. The aim of this study is to assess their suitability for an application of interest today, that of testing of 3D-printed polylactic acid (PLA)-based reactors for biogas production using anaerobic digestion. The impact of temperature, pH, and aqueous phase on the tested bioreactor is investigated, together with the effect of the gaseous phase (i.e., produced biogas). Two batches of materials used separately, one after another inside the bioreactor were considered, in a realistic situation. Two essential parameters inside the reactor (i.e., pH and temperature) were continuously monitored during a time interval of 25 to 30 days for each of the two biogas-generating processes. To understand the impact of these processes on the walls of the bioreactor, samples of 3D-printed material were placed at three levels: at the top (i.e., outside the substrate), in the middle, and at the bottom of the bioreactor. The samples were analyzed using a non-destructive imaging method, Optical Coherence Tomography (OCT). An in-house developed swept-source (SS) OCT system, master-slave (MS) enhanced, operating at a central wavelength of 1310 nm was utilized. The 3D OCT images related to the degradation level of the material of the PLA samples were validated using Scanning Electron Microscopy (SEM). The differences between the impact of the substrate on samples situated at the three considered levels inside the reactor were determined and analyzed using their OCT B-scans (optical cross-section images). Thus, the impact of the biogas-generating process on the interior of the bioreactor was demonstrated and quantified, as well as the capability of OCT to perform such assessments. Therefore, future work may target OCT for in situ investigations of such bioreactors.

Enhanced resolution optoacoustic microscopy using a picosecond high repetition rate Q-switched microchip laser.

Conventional optoacoustic microscopy (OAM) instruments have at their core a nanosecond pulse duration laser. If lasers with a shorter pulse duration are used, broader, higher frequency ultrasound waves are expected to be generated and as a result, the axial resolution of the instrument is improved. Here, we exploit the advantage offered by a picosecond duration pulse laser to enhance the axial resolution of an OAM instrument. In comparison to an instrument equipped with a 2-ns pulse duration laser, an improvement in the axial resolution of 50% is experimentally demonstrated by using excitation pulses of only 85 ps. To illustrate the capability of the instrument to generate high-quality optoacoustic images, en-face, in-vivo images of the brain of Xenopus laevis tadpole are presented with a lateral resolution of 3.8 µ m throughout the entire axial imaging range.

Two octaves spanning photoacoustic microscopy.

In this study, for the first time, a Photoacoustic Microscopy instrument driven by a single optical source operating over a wide spectral range (475-2400 nm), covering slightly more than two octaves is demonstrated. Xenopus laevis tadpoles were imaged in vivo using the whole spectral range of 2000 nm of a supercontinuum optical source, and a novel technique of mapping absorbers is also demonstrated, based on the supposition that only one chromophore contributes to the photoacoustic signal of each individual voxel in the 3D photoacoustic image. By using a narrow spectral window (of 25 nm bandwidth) within the broad spectrum of the supercontinuum source at a time, in vivo hyper-spectral Photoacoustic images of tadpoles are obtained. By post-processing pairs of images obtained using different spectral windows, maps of five endogenous contrast agents (hemoglobin, melanin, collagen, glucose and lipids) are produced.

Endoscopic en-face optical coherence tomography and fluorescence imaging using correlation-based probe tracking.

Forward-viewing endoscopic optical coherence tomography (OCT) provides 3D imaging in vivo, and can be combined with widefield fluorescence imaging by use of a double-clad fiber. However, it is technically challenging to build a high-performance miniaturized 2D scanning system with a large field-of-view. In this paper we demonstrate how a 1D scanning probe, which produces cross-sectional OCT images (B-scans) and 1D fluorescence T-scans, can be transformed into a 2D scanning probe by manual scanning along the second axis. OCT volumes are assembled from the B-scans using speckle decorrelation measurements to estimate the out-of-plane motion along the manual scan direction. Motion within the plane of the B-scans is corrected using image registration by normalized cross correlation. En-face OCT slices and fluorescence images, corrected for probe motion in 3D, can be displayed in real-time during the scan. For a B-scan frame rate of 250 Hz, and an OCT lateral resolution of approximately 20 µ m , the approach can handle out-of-plane motion at speeds of up to 4 mm/s.

Optimization of X-ray Investigations in Dentistry Using Optical Coherence Tomography.

The most common imaging technique for dental diagnoses and treatment monitoring is X-ray imaging, which evolved from the first intraoral radiographs to high-quality three-dimensional (3D) Cone Beam Computed Tomography (CBCT). Other imaging techniques have shown potential, such as Optical Coherence Tomography (OCT). We have recently reported on the boundaries of these two types of techniques, regarding. the dental fields where each one is more appropriate or where they should be both used. The aim of the present study is to explore the unique capabilities of the OCT technique to optimize X-ray units imaging (i.e., in terms of image resolution, radiation dose, or contrast). Two types of commercially available and widely used X-ray units are considered. To adjust their parameters, a protocol is developed to employ OCT images of dental conditions that are documented on high (i.e., less than 10 µm) resolution OCT images (both B-scans/cross sections and 3D reconstructions) but are hardly identified on the 200 to 75 µm resolution panoramic or CBCT radiographs. The optimized calibration of the X-ray unit includes choosing appropriate values for the anode voltage and current intensity of the X-ray tube, as well as the patient's positioning, in order to reach the highest possible X-rays resolution at a radiation dose that is safe for the patient. The optimization protocol is developed in vitro on OCT images of extracted teeth and is further applied in vivo for each type of dental investigation. Optimized radiographic results are compared with un-optimized previously performed radiographs. Also, we show that OCT can permit a rigorous comparison between two (types of) X-ray units. In conclusion, high-quality dental images are possible using low radiation doses if an optimized protocol, developed using OCT, is applied for each type of dental investigation. Also, there are situations when the X-ray technology has drawbacks for dental diagnosis or treatment assessment. In such situations, OCT proves capable to provide qualitative images.

Dental Diagnosis and Treatment Assessments: Between X-rays Radiography and Optical Coherence Tomography.

A correct diagnosis in dental medicine is typically provided only after clinical and radiological evaluations. They are also required for treatment assessments. The aim of this study is to establish the boundaries from which a modern, although established, imaging technique, Optical Coherence Tomography (OCT), is more suitable than the common X-ray radiography to assess dental issues and treatments. The most common methods for daily-basis clinical imaging are utilized in this study for extracted teeth (but also for other dental samples and materials), i.e., panoramic, intraoral radiography, and three-dimensional (3D) cone beam computed tomography (CBCT). The advantages of using OCT as an imaging method in dentistry are discussed, with a focus on its superior image resolution. Drawbacks related to its limited penetration depth and Field-of-View (FOV) are pointed out. High-quality radiological investigations are performed, measurements are done, and data collected. The same teeth and samples are also imaged (mostly) with an in-house developed Swept Source (SS)-OCT system, Master-Slave enhanced. Some of the OCT investigations employed two other in-house developed OCT systems, Spectral Domain (SD) and Time Domain (TD). Dedicated toolbars from Romexis software (Planmeca, Helsinki, Finland) are used to perform measurements using both radiography and OCT. Clinical conclusions are drawn from the investigations. Upsides and downsides of the two medical imaging techniques are concluded for each type of considered diagnosis. For treatment assessments, it is concluded that OCT is more appropriate than radiography in all applications, except bone-related investigations and periodontitis that demand data from higher-penetration depths than possible with the current level of OCT technology.

All-fibre supercontinuum laser for in vivo multispectral photoacoustic microscopy of lipids in the extended near-infrared region.

Among the numerous endogenous biological molecules, information on lipids is highly coveted for understanding both aspects of developmental biology and research in fatal chronic diseases. Due to the pronounced absorption features of lipids in the extended near-infrared region (1650-1850 nm), visualisation and identification of lipids become possible using multi-spectral photoacoustic (optoacoustic) microscopy. However, the spectroscopic studies in this spectral region require lasers that can produce high pulse energies over a broad spectral bandwidth to efficiently excite strong photoacoustic signals. The most well-known laser sources capable of satisfying the multi-spectral photoacoustic microscopy requirements (tunability and pulse energy) are tunable nanosecond optical parametric oscillators. However, these lasers have an inherently large footprint, thus preventing their use in compact microscopy systems. Besides, they exhibit low-repetition rates. Here, we demonstrate a compact all-fibre, high pulse energy supercontinuum laser that covers a spectral range from 1440 to 1870 nm with a 7 ns pulse duration and total energy of 18.3 µJ at a repetition rate of 100 kHz. Using the developed high-pulse energy source, we perform multi-spectral photoacoustic microscopy imaging of lipids, both ex vivo on adipose tissue and in vivo to study the development of Xenopus laevis tadpoles, using six different excitation bands over the first overtone transition of C-H vibration bonds (1650-1850 nm).

Quantitative assessment of rat bone regeneration using complex master-slave optical coherence tomography.

BACKGROUND: The need for hard and soft tissues in oral implantology determined the development of methods and techniques to increase bone volume and their quality with different alternative materials used as substituents of patient's natural bone. In addition, laser radiation can be used to accelerate the repair of fractures and to produce an increased volume of formed callus, as well as an increased bone mineral density. METHODS: The aim of this work is to evaluate the capability of an in-house developed multimodal complex master slave (CMS) enhanced swept source (SS) optical coherence tomography (OCT) imaging instrument to analyze the increase in the quantity and the improvement of the quality of newly-formed bone using low level laser therapy (LLLT). Bone formation is quantitatively assessed in 5 mm cylindrical defects made in the calvaria part of the skull of living rats. Samples are divided in three study groups: A, a negative control group, for which the natural healing process of the defect is investigated; B, a positive control group, for which bovine graft is used to stimulate bone formation, and C, a study group, in which bovine graft is added to the created defects and LLLT is applied throughout the entire healing period. The animals are sacrificed after 14, 21, and 30 days, and the samples are imaged using the multimodal CMS/SS-OCT instrument. RESULTS: The method allows for the simultaneous monitoring of the bone tissue via two perpendicular cross-sections and nine en-face images taken at adjustable depths into the sample. A global image with course axial resolution allows for the positioning of the field-of-view of the system on the area of interest on the tissue. The quantitative assessment of the process of bone formation is completed using the differences in brightness between the native bone, the artificial bone graft, and the newly-formed bone. CONCLUSIONS: Group C is demonstrated to have a higher volume of newly-formed bone than Group B, which is better from this point of view than Group A. By analyzing the evolution of this volume of new bone in time, the most significant difference was after 21 days, therefore approximately after two thirds of the total time interval analyzed. After 30 days, the volumes of bone tend to move closer, as they begin to fill the available gap. The study demonstrates that OCT can assess quantitatively the positive impact of LLLT on bone regeneration.

En-face optical coherence tomography/fluorescence endomicroscopy for minimally invasive imaging using a robotic scanner.

We report a compact rigid instrument capable of delivering en-face optical coherence tomography (OCT) images alongside (epi)-fluorescence endomicroscopy (FEM) images by means of a robotic scanning device. Two working imaging channels are included: one for a one-dimensional scanning, forward-viewing OCT probe and another for a fiber bundle used for the FEM system. The robotic scanning system provides the second axis of scanning for the OCT channel while allowing the field of view (FoV) of the FEM channel to be increased by mosaicking. The OCT channel has resolutions of 25 / 60 µm (axial/lateral) and can provide en-face images with an FoV of 1.6 × 2.7 mm2. The FEM channel has a lateral resolution of better than 8 µm and can generate an FoV of 0.53 × 3.25 mm2 through mosaicking. The reproducibility of the scanning was determined using phantoms to be better than the lateral resolution of the OCT channel. Combined OCT and FEM imaging were validated with ex-vivo ovine and porcine tissues, with the instrument mounted on an arm to ensure constant contact of the probe with the tissue. The OCT imaging system alone was validated for in-vivo human dermal imaging with the handheld instrument. In both cases, the instrument was capable of resolving fine features such as the sweat glands in human dermal tissue and the alveoli in porcine lung tissue.

Fast spectrally encoded Mueller optical scanning microscopy.

Mueller microscopes enable imaging of the optical anisotropic properties of biological or non-biological samples, in phase and amplitude, at sub-micrometre scale. However, the development of Mueller microscopes poses an instrumental challenge: the production of polarimetric parameters must be sufficiently quick to ensure fast imaging, so that the evolution of these parameters can be visualised in real-time, allowing the operator to adjust the microscope while constantly monitoring them. In this report, a full Mueller scanning microscope based on spectral encoding of polarization is presented. The spectrum, collected every 10 µs for each position of the optical beam on the specimen, incorporates all the information needed to produce the full Mueller matrix, which allows simultaneous display of all the polarimetric parameters, at the unequalled rate of 1.5 Hz (for an image of 256 × 256 pixels). The design of the optical blocks allows for the real-time display of linear birefringent images which serve as guidance for the operator. In addition, the instrument has the capability to easily switch its functionality from a Mueller to a Second Harmonic Generation (SHG) microscope, providing a pixel-to-pixel matching of the images produced by the two modalities. The device performance is illustrated by imaging various unstained biological specimens.

Optical Coherence Tomography Investigations and Modeling of the Sintering of Ceramic Crowns.

Dental prostheses are sintered in ovens that sometimes suffer from a loss of calibration. This can lead to variations of the sintering temperature outside the range recommended by the manufacturer. Stress and even fractures in dental ceramics may occur, and this leads to the necessity to rebuild the dental construct. The aim of this work is to monitor the quality of sintering processes using an established biomedical imaging technique-optical coherence tomography (OCT). Conventional current procedures imply the fabrication of supplemental samples that add to the expenses and are only evaluated visually. To our knowledge, we were the first to propose the use of OCT, a non-destructive method that brings objectivity for such assessments, focusing, in a previous study, on metal ceramic dental prostheses. Here, a different material, pressed ceramics, is considered, while we propose a quantitative assessment of the results-using reflectivity profiles of en-face (i.e., constant-depth) OCT images of sintered samples. The results for both the pressed ceramics and metal ceramics prostheses are discussed by obtaining the analytic functions of their reflectivity profiles. A multi-parametric analysis demonstrates the best parameter to characterize the loss of calibration of dental ovens. Rules-of-thumb are extracted; producing dental prostheses with defects can thus be avoided.

Down-conversion en-face optical coherence tomography.

We present an optical coherence tomography (OCT) method that can deliver an en-face OCT image from a sample in real-time, irrespective of the tuning speed of the swept source. The method, based on the master slave interferometry technique, implements a coherence gate principle by requiring that the optical path difference (OPD) between the arms of an imaging interferometer is the same with the OPD in an interrogating interferometer. In this way, a real-time en-face OCT image can originate from a depth in the sample placed in the imaging interferometer, selected by actuating on the OPD in the interrogating interferometer, while laterally scanning the incident beam over the sample. The generation of the en-face image resembles time domain OCT, with the difference that here the signal is processed based on spectral domain OCT. The optoelectronic processor operates down-conversion of the chirped radio frequency signal delivered by the photo-detector. The down-conversion factor is equal to the ratio of the maximum frequency of the photo-detected signal due to an OPD value matching the coherence length of the swept source, to the sweeping rate. This factor can exceed 106 for long coherence swept sources.

Recovering distance information in spectral domain interferometry.

This work evaluates the performance of the Complex Master Slave (CMS) method, that processes the spectra at the interferometer output of a spectral domain interferometry device without involving Fourier transforms (FT) after data acquisition. Reliability and performance of CMS are compared side by side with the conventional method based on FT, phase calibration with dispersion compensation (PCDC). We demonstrate that both methods provide similar results in terms of resolution and sensitivity drop-off. The mathematical operations required to produce CMS results are highly parallelizable, allowing real-time, simultaneous delivery of data from several points of different optical path differences in the interferometer, not possible via PCDC.

Group refractive index and group velocity dispersion measurement by complex master slave interferometry.

This paper demonstrates that the complex master slave interferometry (CMSI) method used in spectral domain interferometry (SDI) can efficiently be used for accurate refractive index and group velocity dispersion measurements of optically transparent samples. For the first time, we demonstrate the relevance of the phase information delivered by CMSI for dispersion evaluations with no need to linearize data. The technique proposed here has been used to accurately measure the group refractive index and the group velocity dispersion of a strong dispersive sample (SF6 glass), and a weak dispersive one (distilled water). The robustness of the technique is demonstrated through the manipulation of several sets of experimental data.

Passive optical module for polarization-sensitive optical coherence tomography systems.

The paper presents a proof-of-concept polarization-sensitive swept source optical coherence tomography (OCT) system that performs measurements of the retardance as well as of the axis orientation of a linear birefringent sample. The system performs single input state polarization-sensitive OCT and employs an optical module based on optically passive elements such as two beam displacers and a Faraday rotator. Our implementation of the PS-OCT system does not need any calibration step to compensate for the polarimetric effect of the fibers, and its operation does not require a balanced polarization-diversity detector. The optical module allows measurement of the two polarization properties of the sample via two measurements which are performed simultaneously.

Optimization of excitation of fiber Fabry-Perot tunable filters used in swept lasers using a phase-correction method.

In this paper we investigate a phase-correction method for compensation of the nonlinearity of conventional wavelength-swept laser sources based on a fiber Fabry-Perot tunable filter as a wavelength selective element. A triangular waveform signal is commonly used to drive the filter. We, however, extract the zero crossings from the interferograms and modify the shape of the triangular signal accordingly. This algorithm was tested for different values of the optical path length difference in the interferometer setup. Significant compensation for the nonlinearity of the filter was obtained.

Gabor fusion master slave optical coherence tomography.

This paper describes the application of the Gabor filtering protocol to a Master/Slave (MS) swept source optical coherence tomography (SS)-OCT system at 1300 nm. The MS-OCT system delivers information from selected depths, a property that allows operation similar to that of a time domain OCT system, where dynamic focusing is possible. The Gabor filtering processing following collection of multiple data from different focus positions is different from that utilized by a conventional swept source OCT system using a Fast Fourier transform (FFT) to produce an A-scan. Instead of selecting the bright parts of A-scans for each focus position, to be placed in a final B-scan image (or in a final volume), and discarding the rest, the MS principle can be employed to advantageously deliver signal from the depths within each focus range only. The MS procedure is illustrated on creating volumes of data of constant transversal resolution from a cucumber and from an insect by repeating data acquisition for 4 different focus positions. In addition, advantage is taken from the tolerance to dispersion of the MS principle that allows automatic compensation for dispersion created by layers above the object of interest. By combining the two techniques, Gabor filtering and Master/Slave, a powerful imaging instrument is demonstrated. The master/slave technique allows simultaneous display of three categories of images in one frame: multiple depth en-face OCT images, two cross-sectional OCT images and a confocal like image obtained by averaging the en-face ones. We also demonstrate the superiority of MS-OCT over its FFT based counterpart when used with a Gabor filtering OCT instrument in terms of the speed of assembling the fused volume. For our case, we show that when more than 4 focus positions are required to produce the final volume, MS is faster than the conventional FFT based procedure.

Master/slave optical coherence tomography imaging of eyelid basal cell carcinoma.

Optical coherence tomography (OCT) is fast emerging as an additional non-interventional modality for skin tumor detection and diagnosis. A master/slave flying spot OCT configuration was assembled to detect periocular basal cell carcinomas (BCC). A swept source at 1300 nm and sweeping speed of 50 kHz were used. A three-step process was involved. First, 384 channeled spectra using a mirror were stored for 384 optical path differences at the master stage. Then, the stored channeled spectra (masks) were correlated with the channeled spectrum from the BCC tissue to produce 384 en face OCT images (200×200 pixels) for the optical path difference values used to acquire the masks. Finally, these en face slices were stacked to form a volume to cross-reference BCC tumor margins in the orthogonal plane. Per each eyelid sample, several en face images of 200×200 lateral pixels are produced in the time to scan laterally a complete raster of 1.6 s. Combination of the en face views with the cross-sectioning views allow for better discrimination of BCCs comparable to using cross-sectional imaging alone, as previously reported using the conventional fast-Fourier-transform-based OCT techniques.

Master/slave interferometry - ideal tool for coherence revival swept source optical coherence tomography.

In this paper, we demonstrate that the master slave (MS) interferometry method can significantly simplify the practice of coherence revival swept source optical coherence tomography (OCT) technique. Previous implementations of the coherence revival technique required considerable resources on dispersion compensation and data resampling. The total tolerance of the MS method to nonlinear tuning, to dispersion in the interferometer and to dispersion due to the laser cavity, makes the MS ideally suited to the practice of coherence revival. In addition, enhanced versatility is allowed by the MS method in displaying shorter axial range images than that determined by the digital sampling of the data. This brings an immediate improvement in the speed of displaying cross-sectional images at high rates without the need of extra hardware such as graphics processing units or field programmable gate arrays. The long axial range of the coherence revival regime is proven with images of the anterior segment of healthy human volunteers.