30 Years of Optical Coherence Tomography: introduction to the feature issue.

The guest editors introduce a feature issue commemorating the 30th anniversary of Optical Coherence Tomography.

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.

En-face optical coherence tomography analysis of gold and silver nanoparticles in endodontic irrigating solutions: An in vitro study.

Optical coherence tomography (OCT) is a non-invasive, non-radioactive optical diagnostic method based on low-coherence interferometry, which achieves images with different orientation. In dentistry, its major advantage is represented by the localization and characterization of the smallest defects in hard dental tissues, dental materials and of the smallest details in dental anatomy (supplementary canals, recesses, isthmuses, or intra-radicular connections). The aim of the present in vitro study was to evaluate using c-scan en-face optical coherence tomography, the optical opacity, and the distribution inside the root canal lumen of several extracted human teeth of silver and gold nanoparticles from special irrigating solutions used in endodontic treatment. Twelve root canals from 5 human teeth were instrumented using the ProTaper Universal system after initial negotiation with hand K-files ISO no. 10 and rotary nickel-titanium PathFile instruments. An initial c-scan OCT analysis was performed for each sample to confirm that the root canal lumen was empty from radiopaque materials (Group 1). Teeth were first irrigated with NanoCare Plus (Group 2) and then with NanoCare Gold (Group 3) and C-scans were repeated after each irrigation method. The OCT investigation started at the tooth apex, at a depth of 1 mm from its tip. Subsequently, 100 slices of 10 microns were obtained from each root canal. Images were captured and then analyzed with ImageJ software to calculate the level of grey inside the root canal lumen. The highest values of grey were obtained in the samples irrigated with NanoCare Gold after NanoCare Plus (Group 3). The present study proved that both nanoparticles inserted in root canal irrigants were evidenced through OCT imagistic analysis due to their optical opacity, which allowed their highlighting in an empty root canal lumen, after the endodontic treatment was performed and the root canal was cleaned and shaped using specific protocols.

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.

Nanomedicines and microneedles: a guide to their analysis and application.

The fast-advancing progress in the research of nanomedicine and microneedle applications in the past two decades has suggested that the combination of the two concepts could help to overcome some of the challenges we are facing in healthcare. They include poor patient compliance with medication and the lack of appropriate administration forms that enable the optimal dose to reach the target site. Nanoparticles as drug vesicles can protect their cargo and deliver it to the target site, while evading the body's defence mechanisms. Unfortunately, despite intense research on nanomedicine in the past 20 years, we still haven't answered some crucial questions, e.g. about their colloidal stability in solution and their optimal formulation, which makes the translation of this exciting technology from the lab bench to a viable product difficult. Dissolvable microneedles could be an effective way to maintain and stabilise nano-sized formulations, whilst enhancing the ability of nanoparticles to penetrate the stratum corneum barrier. Both concepts have been individually investigated fairly well and many analytical techniques for tracking the fate of nanomaterials with their precious cargo, both in vitro and in vivo, have been established. Yet, to the best of our knowledge, a comprehensive overview of the analytical tools encompassing the concepts of microneedles and nanoparticles with specific and successful examples is missing. In this review, we have attempted to briefly analyse the challenges associated with nanomedicine itself, but crucially we provide an easy-to-navigate scheme of methods, suitable for characterisation and imaging the physico-chemical properties of the material matrix.

Sub-surface characterisation of latest-generation identification documents using optical coherence tomography.

The identification of individuals, particularly at international border crossings, coupled with the evolving sophistication of identity documents are issues that authorities must contend with. Particularly, the ability to distinguish legitimate from counterfeit documents, with high throughput, sensitivity, and selectivity is an ever-evolving challenge. Over the last decade, an increasing number of security features have been introduced by authorities in identification documents. The latest generation of travel documents (such as passports and national ID cards) forego paper substrates for several layers of polycarbonate, allowing security features to be embedded within the documents. These security features may contain information at either the superficial and sub-surface levels, thus increasing the document's resilience to counterfeiting. As the documents become harder to forge, so does the sophistication of forgery detection. There appears to be an unmet and evolving need to identify such sophisticated forgeries, in a non-destructive, high throughput manner. In this publication, we report on the application of optical coherence tomography (OCT) imaging on assessing security features in specimen passports and national ID cards. OCT allows sub-surface imaging of translucent structures, non-destructively enabling quantitative visualisation of embedded security features.

Endodontic fillings evaluated using en face OCT, microCT and SEM.

OBJECTIVES: The goal of endodontic therapy is mechanically cleaning and shaping the root canal system, the removal of organic and inorganic debris followed by sealing with permanent filling materials. MATERIALS AND METHODS: Therefore, the aim of this in vitro study was to engage three imagistic methods: (i) en face (ef) time domain (TD) optical coherence tomography (OCT), (ii) micro-computed tomography (µCT), and (iii) scanning electron microscopy (SEM), in terms of their efficiency in assessing the quality of endodontic fillings. So far, is settled that efOCT images can identify defects∕voids in several of the investigated root canal fillings and identify gaps of 50 µm. RESULTS: The results delivered by µCT technology also showed several imperfections of the endodontic filling but also at the interfaces formed by the sealer and the root canal walls. SEM investigations highlights the complex form of the interface formed the endodontic filling material and the walls of the root canal, as well as shortcomings of the materials from several samples. Gaps of 50 µm are identified with efOCT. CONCLUSIONS: The net advantage of OCT technology, in respect to the other two technologies consists in its non-invasiveness. The OCT axial resolution is also sufficient to see the material gaps. Another advantage of efOCT investigations is that they allow real-time imaging of complex arrangement at the interfaces of the filling material with dentinal root.

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).

Development of Gold-PAGE: towards the electrophoretic analysis of sulphurous biopolymers.

The prevalence, distinctive reactivity and biological significance of sulphur-based groups in proteins and nucleic acids means that analysis of sulphur is of prime importance in biochemistry, biotechnology, and medicine. We report steps in the development of a method to aid in the detection of these moieties using gold nanoparticles as adjuncts in polyacrylamide gel electrophoresis (Gold-PAGE).

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.

Progress in Multimodal En Face Imaging: feature introduction.

This feature issue contains papers that report on the most recent advances in the field of en face optical coherence tomography (OCT) and of combinations of modalities facilitated by the en face view. Hardware configurations for delivery of en face OCT images are described as well as specific signal and image processing techniques tailored to deliver relevant clinical diagnoses. The value of the en face perspective for enabling multimodality is illustrated by several combination modalities.

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.

Real-time high-resolution mid-infrared optical coherence tomography.

The potential for improving the penetration depth of optical coherence tomography systems by using light sources with longer wavelengths has been known since the inception of the technique in the early 1990s. Nevertheless, the development of mid-infrared optical coherence tomography has long been challenged by the maturity and fidelity of optical components in this spectral region, resulting in slow acquisition, low sensitivity, and poor axial resolution. In this work, a mid-infrared spectral-domain optical coherence tomography system operating at a central wavelength of 4 µm and an axial resolution of 8.6 µm is demonstrated. The system produces two-dimensional cross-sectional images in real time enabled by a high-brightness 0.9- to 4.7-µm mid-infrared supercontinuum source with a pulse repetition rate of 1 MHz for illumination and broadband upconversion of more than 1-µm bandwidth from 3.58-4.63 µm to 820-865 nm, where a standard 800-nm spectrometer can be used for fast detection. The images produced by the mid-infrared system are compared with those delivered by a state-of-the-art ultra-high-resolution near-infrared optical coherence tomography system operating at 1.3 µm, and the potential applications and samples suited for this technology are discussed. In doing so, the first practical mid-infrared optical coherence tomography system is demonstrated, with immediate applications in real-time non-destructive testing for the inspection of defects and thickness measurements in samples that exhibit strong scattering at shorter wavelengths.