Two optical coherence tomography systems detect topical gold nanoshells in hair follicles,sweat ducts and measure epidermis

Optical coherence tomography (OCT) is an established imaging technology for in vivo skin investigation. Topical application of gold nanoshells (GNS) provides contrast enhancement in OCT by generating a strong hyperreflective signal from hair follicles and sweat glands, which are the natural skin openings. This study explores the utility of 150 nm diameter GNS as contrast agent for OCT imaging. GNS was massaged into skin and examined in four skin areas of 11 healthy volunteers. A commercial OCT system and a prototype with 3 μm resolution (UHR‐OCT) were employed to detect potential benefits of increased resolution and variability in intensity generated by the GNS. In both OCT‐systems GNS enhanced contrast from hair follicles and sweat ducts. Highest average penetration depth of GNS was in armpit 0.64 mm ± SD 0.17, maximum penetration depth was 1.20 mm in hair follicles and 15 to 40 μm in sweat ducts. Pixel intensity generated from GNS in hair follicles was significantly higher in UHR‐OCT images (P = .002) and epidermal thickness significantly lower 0.14 vs 0.16 mm (P = .027). This study suggests that GNSs are interesting candidates for increasing sensitivity in OCT diagnosis of hair and sweat gland disorders and demonstrates that choice of OCT systems influences results.      

Depth-dependent in vivo human skin backscattering spectra extraction from full-field optical coherence tomography

With homemade active crystalline fibers, we generated bright and broadband light sources for full-field optical coherence tomography, offering deep penetration into skin tissues with cellular resolution at a high frame rate. Extraction of backscattered spectra from the tissue has potential applications in biomedicine. The hysteresis nonlinearity of the piezoelectric transducer actuating the Mirau interferometer has been greatly reduced by a feedforward compensation approach. The linearized hysteresis response enables us to extract depth-dependent spectra accurately. To validate, the complex dispersion of a fused silica plate was characterized with 2% error. Further validation on an in vitro setting, the backscattered spectra from indocyanine green pigment and nonpigmented microspheres were obtained and verified. For in vivo skin measurement, the backscattered spectra show depth-dependent spectral shift and bandwidth variation due to the complex skin anatomy and pigment absorption. Such a high-speed spectra acquisition of in vivo deep tissue backscattering could lead to disease diagnosis in clinical settings.     

Investigation of the human tympanic membrane oscillation ex vivo by Doppler optical coherence tomography

Investigations of the tympanic membrane (TM) can have an important impact on understanding the sound conduction in the ear and can therefore support the diagnosis and treatment of diseases in the middle ear. High‐speed Doppler optical coherence tomography (OCT) has the potential to describe the oscillatory behaviour of the TM surface in a phase‐sensitive manner and additionally allows acquiring a three‐dimensional image of the underlying structure. With repeated sound stimuli from 0.4 kHz to 6.4 kHz, the whole TM can be set in vibration and the spatially resolved frequency response functions (FRFs) of the tympanic membrane can be recorded. Typical points, such as the umbo or the manubrium of malleus, can be studied separately as well as the TM surface with all stationary and wave‐like vibrations. Thus, the OCT methodology can be a promising technique to distinguish between normal and pathological TMs and support the differentiation between ossicular and membrane diseases. (© 2014 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

In vivo monitoring of thrombosis in mice by optical coherence tomography

The objective of this study is to establish a novel method for continuously monitoring thrombus progression with various outcome measures and to assess the efficacy of antithrombotic drugs in murine thrombosis model in mice. In the study, thrombus was induced in the femoral vein of mice by FeCl₃ and monitored over time by spectral‐domain optical coherence tomography (OCT). Three‐dimensional images of thrombi with or without heparin as an antithrombotic agent were obtained from OCT angiography. In addition, several parameters of thrombi were analyzed and compared between control and anticoagulant groups. By using OCT, we were able to trace thrombus generation in the same mouse in real time. We found that in our model heparin reduced thrombus size by ~60% and thrombus cross‐sectional area by 50%. OCT results also show that both time to thrombus size (>0.02mm³) and time to occlusion (>30%) were significantly reduced after heparin addition. This study demonstrates that OCT reliably monitors thrombus generation and progression from various aspects including thrombus size. This enables us to measure the kinetic of thrombosis more accurately, and effectively evaluate the efficacy and activities of antithrombotic drugs. This model may represent a useful tool in antithrombotic drug discoveries in preclinical studies.      

Pilot feasibility study of in vivo intraoperative quantitative optical coherence tomography of human brain tissue during glioma resection

This study investigates the feasibility of in vivo quantitative optical coherence tomography (OCT) of human brain tissue during glioma resection surgery in six patients. High‐resolution detection of glioma tissue may allow precise and thorough tumor resection while preserving functional brain areas, and improving overall survival. In this study, in vivo 3D OCT datasets were collected during standard surgical procedure, before and after partial resection of the tumor, both from glioma tissue and normal parenchyma. Subsequently, the attenuation coefficient was extracted from the OCT datasets using an automated and validated algorithm. The cortical measurements yield a mean attenuation coefficient of 3.8 ± 1.2 mm^?1 for normal brain tissue and 3.6 ± 1.1 mm^?1 for glioma tissue. The subcortical measurements yield a mean attenuation coefficient of 5.7 ± 2.1 and 4.5 ± 1.6 mm^?1 for, respectively, normal brain tissue and glioma. Although the results are inconclusive with respect to trends in attenuation coefficient between normal and glioma tissue due to the small sample size, the results are in the range of previously reported values. Therefore, we conclude that the proposed method for quantitative in vivo OCT of human brain tissue is feasible during glioma resection surgery.     

In vivo dynamic characterization of the human tympanic membrane using pneumatic optical coherence tomography

Decreased mobility of the human eardrum, the tympanic membrane (TM), is an essential indicator of a prevalent middle ear infection. The current diagnostic method to assess TM mobility is via pneumatic otoscopy, which provides subjective and qualitative information of subtle motion. In this study, a handheld spectral-domain pneumatic optical coherence tomography system was developed to simultaneously measure the displacement of the TM, air pressure inputs applied to a sealed ear canal, and to perform digital pneumatic otoscopy. A novel approach based on quantitative parameters is presented to characterize spatial and temporal variations of the dynamic TM motion. Furthermore, the TM motions of normal middle ears are compared with those of ears with middle ear infections. The capability of noninvasively measuring the rapid motion of the TM is beneficial to understand the complex dynamics of the human TM, and can ultimately lead to improved diagnosis and management of middle ear infections.     

Characterization of the human myocardium by optical coherence tomography

Imaging of cardiac tissue structure plays a critical role in the treatment and understanding of cardiovascular disease. Optical coherence tomography (OCT) offers the potential to provide valuable, high‐resolution imaging of cardiac tissue. However, there is a lack of comprehensive OCT imaging data of the human heart, which could improve identification of structural substrates underlying cardiac abnormalities. The objective of this study was to provide qualitative and quantitative analysis of OCT image features throughout the human heart. Fifty human hearts were acquired, and tissues from all chambers were imaged with OCT. Histology was obtained to verify tissue composition. Statistical differences between OCT image features corresponding to different tissue types and chambers were estimated using analysis of variance. OCT imaging provided features that were able to distinguish structures such as thickened collagen, as well as adipose tissue and fibrotic myocardium. Statistically significant differences were found between atria and ventricles in attenuation coefficient, and between adipose and all other tissue types. This study provides an overview of OCT image features throughout the human heart, which can be used for guiding future applications such as OCT‐integrated catheter‐based treatments or ex vivo investigation of structural substrates.     

Cross‐scanning optical coherence tomography angiography for eye motion correction

We propose a cross‐scanning optical coherence tomography (CS‐OCT) system to correct eye motion artifacts in OCT angiography images. This system employs a dual‐illumination configuration with two orthogonally polarized beams, each of which simultaneously perform raster scanning in perpendicular direction with each other over the same area. In the reference arm, a polarization delay unit is used to acquire the two orthogonally polarized interferograms with a single photo detector by introducing different optical delay lines. The two cross‐scanned volume data are affected by the same eye motion but in two orthogonal directions. We developed a motion correction algorithm, which removes artifacts in the slow axis of each angiogram using the other and merges them through a nonrigid registration algorithm. In this manner, we obtained a motion‐corrected angiogram within a single volume scanning time without additional eye‐tracking devices.     

Hyperspectral optical coherence tomography for in vivo visualization of melanin in the retinal pigment epithelium

Previous studies for melanin visualization in the retinal pigment epithelium (RPE) have exploited either its absorption properties (using photoacoustic tomography or photothermal optical coherence tomography [OCT]) or its depolarization properties (using polarization sensitive OCT). However, these methods are only suitable when the melanin concentration is sufficiently high. In this work, we present the concept of hyperspectral OCT for melanin visualization in the RPE when the concentration is low. Based on white light OCT, a hyperspectral stack of 27 wavelengths (440‐700 nm) was created in post‐processing for each depth‐resolved image. Owing to the size and shape of the melanin granules in the RPE, the variations in backscattering coefficient as a function of wavelength could be identified—a result which is to be expected from Mie theory. This effect was successfully identified both in eumelanin‐containing phantoms and in vivo in the low‐concentration Brown Norway rat RPE.      

In vivo pump‐probe optical coherence tomography imaging in Xenopus laevis

Currently, optical coherence tomography (OCT), is not capable of obtaining molecular information often crucial for identification of disease. To enable molecular imaging with OCT, we have further developed a technique that harnesses transient changes in light absorption in the sample to garner molecular information. A Fourier‐domain Pump‐Probe OCT (PPOCT) system utilizing a 532 nm pump and 830 nm probe has been developed for imaging hemoglobin. Methylene blue, a biological dye with well‐know photophysics, was used to characterize the system before investigating the origin of the hemoglobin PPOCT signal. The first in vivo PPOCT images were recorded of the vasculature in Xenopus laevis. The technique was shown to work equally well in flowing and nonflowing vessels. Furthermore, PPOCT was compared with other OCT extensions which require flow, such as Doppler OCT and phase‐variance OCT. PPOCT was shown to better delineate tortuous vessels, where nodes often restrict Doppler and phase‐variance reconstruction. (© 2013 WILEY‐VCH Verlag GmbH &Co. KGaA, Weinheim)     

Quantifying dental biofilm growth using cross-polarization optical coherence tomography

Aims: Quantifying the ex vivo growth of complex multispecies dental biofilms using cross‐polarization 1310‐nm optical coherence tomography (CP‐OCT) system was investigated. Methods and Results: Bacterial microcosms, which were derived from plaque samples of paediatric subjects, were incubated in a biofilm reactor system containing discs of different dental materials for 72 h with daily sucrose pulsing (5×). CP‐OCT analysis of biofilm mass was validated with crystal violet (CV) assays at various growth stages of these complex biofilms. CP‐OCT was able to filter out the back‐reflected signals of water layers in the hydrated biofilm and allowed for direct biofilm quantification. The overall depth‐resolved scattering intensity of the biofilm showed very strong positive correlation with CV assay quantification (Spearman’s ρ = 0·92) during the growth phase of the biofilm. Conclusion: CP‐OCT was able to quantify the mass of the biofilm by measuring the overall depth‐resolved scattering of the biofilm. Significance and Impact of the Study: CP‐OCT has the ability to nondestructively monitor biofilm growth and elucidate the growth characteristics of these microcosms on different dental material compositions.     

Measuring polarization changes in the human outer retina with polarization‐sensitive optical coherence tomography

Morphological changes in the outer retina such as drusen are established biomarkers to diagnose age‐related macular degeneration. However, earlier diagnosis might be possible by taking advantage of more subtle changes that accompany tissues that bear polarization‐altering properties. To test this hypothesis, we developed a method based on polarization‐sensitive optical coherence tomography with which volumetric data sets of the macula were obtained from 10 young (<25 years) and 10 older (>54 years) subjects. All young subjects and 5 of the older subjects had retardance values induced by the retinal pigment epithelium and Bruch's membrane (RPE‐BM) complex that were just above the noise floor measurement (5°‐13° at 840 nm). In contrast, elevated retardance, up to 180°, was observed in the other 5 older subjects. Analysis of the degree of polarization uniformity (DOPU) demonstrates that reduced DOPU (<0.4) in the RPE is associated with elevated double pass phase retardation (DPPR) below the RPE‐BM complex, suggesting that the observed elevated DPPR in older subjects is the result of increased scattering or polarization scrambling. Collectively, our measurements show that the outer retina can undergo dramatic change in its polarization properties with age, and in some cases still retain its clinically normal appearance.      

Improving cerebral microvascular image quality of optical coherence tomography angiography with deep learning-based segmentation

Optical coherence tomography angiography (OCTA) can map the microvascular networks of the cerebral cortices with micrometer resolution and millimeter penetration. However, the high scattering of the skull and the strong noise in the deep imaging region will distort the vasculature projections and decrease the OCTA image quality. Here, we proposed a deep learning-based segmentation method based on a U-Net convolutional neural network to extract the cortical region from the OCT image. The vascular networks were then visualized by three OCTA algorithms. The image quality of the vasculature projections was assessed by two metrics, including the peak signal-to-noise ratio (PSNR) and the contrast-to-noise ratio (CNR). The results show the accuracy of the cortical segmentation was 96.07%. The PSNR and CNR values increased significantly in the projections of the selected cortical regions. The OCTA incorporating the deep learning-based cortical segmentation can efficiently improve the image quality and enhance the vasculature clarity.     

The AC conductivity of human sweat ducts as the dominant factor in the sub-THz reflection coefficient of skin

The helical nature of human sweat ducts, combined with the morphological and dielectric properties of skin, suggests electromagnetic activity in the sub-THz frequency band. A detailed electromagnetic simulation model of the skin, with embedded sweat ducts, was created. The model includes realistic dielectric properties based on the measured water content of each layer of skin, derived from Raman Spectroscopy. The model was verified by comparing it to measurements of the reflection coefficient of the palms of 13 volunteers in the frequency band 350–410 GHz. They were subjected to a measurement protocol intended to induce mental stress, thereby also activating the sweat glands. The Galvanic Skin Response was concurrently measured. Using the simulation model the optimal ac-conductivity for each measurement was found. The range of variation for all subjects was found to be from 100 S/m to a maximum value of 6000 S/m with averages of 1000 S/m. These are one order of magnitude increase from the accepted values for water at these frequencies (~100 s/m at 100 GHz). Considering the known biochemical mechanism for inducing perspiration, we conclude that these ac-conductivity levels are probably valid, even though the real time measurements of sweat ac-conductivity levels inside the duct are inaccessible.     

Comparative study of optical coherence tomography angiography algorithms for rodent retinal imaging

Optical coherence tomography angiography (OCTA) is a functional extension of optical coherence tomography for non-invasive in vivo three-dimensional imaging of the microvasculature of biological tissues. Several algorithms have been developed to construct OCTA images from the measured optical coherence tomography signals. In this study, we compared the performance of three OCTA algorithms that are based on the variance of phase, amplitude, and the complex representations of the optical coherence tomography signals for rodent retinal imaging, namely the phase variance, improved speckle contrast, and optical microangiography. The performance of the different algorithms was evaluated by comparing the quality of the OCTA images regarding how well the vasculature network can be resolved. Quantities that are widely used in ophthalmic studies including blood vessel density, vessel diameter index, vessel perimeter index, vessel complexity index were also compared. Results showed that both the improved speckle contrast and optical microangiography algorithms are more robust than phase variance, and they can reveal similar vasculature features while there are statistical differences in the calculated quantities.     

General model for depth‐resolved estimation of the optical attenuation coefficients in optical coherence tomography

We present the proof of concept of a general model that uses the tissue sample transmittance as input to estimate the depth‐resolved attenuation coefficient of tissue samples using optical coherence tomography (OCT). This method allows us to obtain an image of tissue optical properties instead of intensity contrast, guiding diagnosis and tissues differentiation, extending its application from thick to thin samples. The performance of our method was simulated and tested with the assistance of a home built single‐layered and multilayered phantoms (~100 μm each layer) with known attenuation coefficient on the range of 0.9 to 2.32 mm^?1. It is shown that the estimated depth‐resolved attenuation coefficient recovers the reference values, measured by using an integrating sphere followed by the inverse adding doubling processing technique. That was corroborated for all situations when the correct transmittance value is used with an average difference of 7%. Finally, we applied the proposed method to estimate the depth‐resolved attenuation coefficient for a thin biological sample, demonstrating the ability of our method on real OCT images.     

Multimodal Raman spectroscopy and optical coherence tomography for biomedical analysis

Optical techniques hold great potential to detect and monitor disease states as they are a fast, non-invasive toolkit. Raman spectroscopy (RS) in particular is a powerful label-free method capable of quantifying the biomolecular content of tissues. Still, spontaneous Raman scattering lacks information about tissue morphology due to its inability to rapidly assess a large field of view. Optical Coherence Tomography (OCT) is an interferometric optical method capable of fast, depth-resolved imaging of tissue morphology, but lacks detailed molecular contrast. In many cases, pairing label-free techniques into multimodal systems allows for a more diverse field of applications. Integrating RS and OCT into a single instrument allows for both structural imaging and biochemical interrogation of tissues and therefore offers a more comprehensive means for clinical diagnosis. This review summarizes the efforts made to date toward combining spontaneous RS-OCT instrumentation for biomedical analysis, including insights into primary design considerations and data interpretation.     

In vivo observation of metamorphosis of Plodia interpunctella Hübner using three‐dimensional optical coherence tomography

Morphological assessment and three‐dimensional reconstructions of internal structures of Plodia interpunctella Hübner during metamorphosis stages were experimentally demonstrated using optical coherence tomography (OCT) for the first time. The conventional, complex sectioning methods were significantly simplified owing to the non‐invasive three‐dimensional imaging capability of OCT. Further, this study demonstrates the use of OCT as a non‐invasive detection tool for in vivo morphological observation of metamorphosis stages to gain a better understanding about the growth of internal organs, which can be considered a useful discovery in the field of entomology. Thus, the metamorphosis stages starting from the larva, three pupa stages to the adult stage were periodically visualized to examine the development of internal organs at each specific stage. This study essentially offers real‐time morphological information by non‐destructive observation of the organism and can also be useful for the investigation of other agricultural pests.     

Quantitative assessment of optical coherence tomography angiography algorithms for neuroimaging

Optical coherence tomography (OCT) angiography can noninvasively map microvascular networks and quantify blood flow in a cerebral cortex with a resolution of 1 to 10 μm and a penetration depth of 2 to 3 mm incorporating OCT signals and angiography algorithms. Different angiography algorithms have been developed in recent years; however, the performance of the algorithms has not been assessed quantitatively for neuroimaging applications. In this paper, we developed four metrics including vascular connectivity, contrast‐to‐noise ratio, signal‐to‐noise ratio and processing time to quantitatively assess the performance of OCT angiography algorithms in image quality and computation speed. After the imaging of a rat cortex using an OCT system, the cerebral microvascular networks were visualized by seven algorithms, and the performance of the algorithms was quantified and compared. Quantitative performance assessment of the algorithms can provide suggestions for the selection of appropriate OCT angiography algorithms in neuroimaging.     

Imaging atherosclerotic plaque composition with intracoronary optical coherence tomography

Optical coherence tomography (OCT) allows highly accurate diagnosis of atherosclerotic plaques, including measurement of the thickness of fibrous caps, permitting an assessment of the risk of rupture. While the OCT image presents morphological information in highly resolved detail, it relies on interpretation by trained readers for the identification of tissue type. We developed a method for quantitative classification of atherosclerotic plaque constituents. The optical attenuation coefficient μ_t distinguishes different tissue types: necrotic core and macrophage infiltration exhibit strong attenuation, μ_t≥10 mm⁻¹, while calcific and fibrous tissue have a lower μ_t≈2–5 mm⁻¹. (Neth Heart J 2009;17:448-50.)