Growing Trend to Adopt Speckle Variance Optical Coherence Tomography for Biological Tissue Assessments in Pre-Clinical Applications.

Speckle patterns are a generic feature in coherent imaging techniques like optical coherence tomography (OCT). Although speckles are granular like noise texture, which degrades the image, they carry information that can be benefited by processing and thereby furnishing crucial information of sample structures, which can serve to provide significant important structural details of samples in in vivo longitudinal pre-clinical monitoring and assessments. Since the motions of tissue molecules are indicated through speckle patterns, speckle variance OCT (SV-OCT) can be well-utilized for quantitative assessments of speckle variance (SV) in biological tissues. SV-OCT has been acknowledged as a promising method for mapping microvasculature in transverse-directional blood vessels with high resolution in micrometers in both the transverse and depth directions. The fundamental scope of this article reviews the state-of-the-art and clinical benefits of SV-OCT to assess biological tissues for pre-clinical applications. In particular, focus on precise quantifications of in vivo vascular response, therapy assessments, and real-time temporal vascular effects of SV-OCT are primarily emphasized. Finally, SV-OCT-incorporating pre-clinical techniques with high potential are presented for future biomedical applications.

A long-term storable gel-laden chip composite built in a multi-well plate enablingin situcell encapsulation for high-throughput liver model.

Hydrogels are widely used as scaffold materials for constructingin vitrothree-dimensional microphysiological systems. However, their high sensitivity to various external cues hinders the development of hydrogel-laden, microscale, and high-throughput chips. Here, we have developed a long-term storable gel-laden chip composite built in a multi-well plate, which enablesin situcell encapsulation and facilitates high-throughput analysis. Through optimized chemical crosslinking and freeze-drying method (C/FD), we have achieved a high-quality of gel-laden chip composite with excellent transparency, uniform porosity, and appropriate swelling and mechanical characteristics. Besides collagen, decellularized extracellular matrix with tissue-specific biochemical compound has been applied as chip composite. As a ready-to-use platform,in situcell encapsulation within the gel has been achieved through capillary force generated during gel reswelling. The liver-mimetic chip composite, comprising HepG2 cells or primary hepatocytes, has demonstrated favorable hepatic functionality and high sensitivity in drug testing. The developed fabrication process with improved stability of gels and storability allows chip composites to be stored at a wide range of temperatures for up to 28 d without any deformation, demonstrating off-the-shelf products. Consequently, this provides an exceptionally simple and long-term storable platform that can be utilized for an efficient tissue-specific modeling and various biomedical applications.

Spectrally encoded dual-mode interferometry with orthogonal scanning.

Spectrally encoded confocal microscopy (SECM) is a high-speed reflectance confocal microscopy technique. Here, we present a method to integrate optical coherence tomography (OCT) and SECM for complementary imaging by adding orthogonal scanning to the SECM configuration. The co-registration of SECM and OCT is automatic, as all system components are shared in the same order, eliminating the need for additional optical alignment. The proposed multimode imaging system is compact and cost-effective while providing the benefits of imaging aiming and guidance. Furthermore, speckle noise can be suppressed by averaging the speckles generated by shifting the spectral-encoded field in the direction of dispersion. Using a near infrared (NIR) card and a biological sample, we demonstrated the capability of the proposed system by showing SECM imaging at depths of interest guided by the OCT in real time and speckle noise reduction. Interfaced multimodal imaging of SECM and OCT was implemented at a speed of approximately 7 frames/s using fast-switching technology and GPU processing.

Label-free photothermal optical coherence microscopy to locate desired regions of interest in multiphoton imaging of volumetric specimens.

Biochip-based research is currently evolving into a three-dimensional and large-scale basis similar to the in vivo microenvironment. For the long-term live and high-resolution imaging in these specimens, nonlinear microscopy capable of label-free and multiscale imaging is becoming increasingly important. Combination with non-destructive contrast imaging will be useful for effectively locating regions of interest (ROI) in large specimens and consequently minimizing photodamage. In this study, a label-free photothermal optical coherence microscopy (OCM) serves as a new approach to locate the desired ROI within biological samples which are under investigation by multiphoton microscopy (MPM). The weak photothermal perturbation in sample by the MPM laser with reduced power was detected at the endogenous photothermal particles within the ROI using the highly sensitive phase-differentiated photothermal (PD-PT) OCM. By monitoring the temporal change of the photothermal response signal of the PD-PT OCM, the hotspot generated within the sample focused by the MPM laser was located on the ROI. Combined with automated sample movement in the x-y axis, the focal plane of MPM could be effectively navigated to the desired portion of a volumetric sample for high-resolution targeted MPM imaging. We demonstrated the feasibility of the proposed method in second harmonic generation microscopy using two phantom samples and a biological sample, a fixed insect on microscope slide, with dimensions of 4 mm wide, 4 mm long, and 1 mm thick.

On-field optical imaging data for the pre-identification and estimation of leaf deformities.

Visually nonidentifiable pathological symptoms at an early stage are a major limitation in agricultural plantations. Thickness reduction in palisade parenchyma (PP) and spongy parenchyma (SP) layers is one of the most common symptoms that occur at the early stage of leaf diseases, particularly in apple and persimmon. To visualize variations in PP and SP thickness, we used optical coherence tomography (OCT)-based imaging and analyzed the acquired datasets to determine the threshold parameters for pre-identifying and estimating persimmon and apple leaf abnormalities using an intensity-based depth profiling algorithm. The algorithm identified morphological differences between healthy, apparently-healthy, and infected leaves by applying a threshold in depth profiling to classify them. The qualitative and quantitative results revealed changes and abnormalities in leaf morphology in addition to disease incubation in both apple and persimmon leaves. These can be used to examine how initial symptoms are influenced by disease growth. Thus, these datasets confirm the significance of OCT in identifying disease symptoms nondestructively and providing a benchmark dataset to the agriculture community for future reference.

Micron-scale human enamel layer characterization after orthodontic bracket debonding by intensity-based layer segmentation in optical coherence tomography images.

In clinical orthodontic practice, fixed brackets are widely used for tooth movement and adjustments. Although years of research and development have improved the workability of fixed orthodontic brackets, there are still controversies regarding its plausible destructive influence on the enamel surface of tooth. This, in turn, makes the quantitative assessment of the enamel surface after specific orthodontic treatment procedures important in order to opt for the most effective treatment procedure. Through this study, we show the practical applicability of optical coherence tomography (OCT) as a non-ionizing and nondestructive assessment tool for measuring enamel loss after each step of orthodontic bracket bonding. Two-dimensional and volumetric OCT images are used for the evaluation of the tooth enamel. From the depth intensity profile analysis of cross-sectional OCT images, the changes in the individual internal layer thickness are calculated. A software algorithm was developed to evaluate the structural connectivity in the enamel for analyzing enamel loss on the tooth surface and for detecting enamel abrasion. An intensity-based layer segmentation algorithm is also developed to analyze and evaluate enamel wear in the tooth after each step. Using the proposed algorithms, the total enamel present after each treatment procedure was measured and tabulated for analysis.

Integrated Quad-Scanner Strategy-Based Optical Coherence Tomography for the Whole-Directional Volumetric Imaging of a Sample.

Whole-directional scanning methodology is required to observe distinctive features of an entire physical structure with a three dimensional (3D) visualization. However, the implementation of whole-directional scanning is challenging for conventional optical coherence tomography (OCT), which scans a limited portion of the sample by utilizing unidirectional and bidirectional scanning methods. Therefore, in this paper an integrated quad-scanner (QS) strategy-based OCT method was implemented to obtain the whole-directional volumetry of a sample by employing four scanning arms installed around the sample. The simultaneous and sequential image acquisition capabilities are the conceptual key points of the proposed QS-OCT method, and were implemented using four precisely aligned scanning arms and applied in a complementary way according to the experimental criteria. To assess the feasibility of obtaining whole-directional morphological structures, a roll of Scotch tape, an ex vivo mouse heart, and kidney specimens were imaged and independently obtained tissue images at different directions were delicately merged to compose the 3D volume data set. The results revealed the potential merits of QS-OCT-based whole-directional imaging, which can be a favorable inspection method for various discoveries that require the dynamic coordinates of the whole physical structure.

In Situ Characterization of Micro-Vibration in Natural Latex Membrane Resembling Tympanic Membrane Functionally Using Optical Doppler Tomography.

Non-invasive characterization of micro-vibrations in the tympanic membrane (TM) excited by external sound waves is considered as a promising and essential diagnosis in modern otolaryngology. To verify the possibility of measuring and discriminating the vibrating pattern of TM, here we describe a micro-vibration measurement method of latex membrane resembling the TM. The measurements are obtained with an externally generated audio stimuli of 2.0, 2.2, 2.8, 3.1 and 3.2 kHz, and their respective vibrations based tomographic, volumetric and quantitative evaluations were acquired using optical Doppler tomography (ODT). The micro oscillations and structural changes which occurred due to diverse frequencies are measured with sufficient accuracy using a highly sensitive ODT system implied phase subtraction method. The obtained results demonstrated the capability of measuring and analyzing the complex varying micro-vibration of the membrane according to implied sound frequency.

Analysis of Enamel Loss by Prophylaxis and Etching Treatment in Human Tooth Using Optical Coherence Tomography: An In Vitro Study.

Bonding of braces is an essential part in contemporary orthodontic treatment. For the proper strength of bracket bonding, enamel conditioning or surface treatment on tooth surface is required. Treatment on the tooth surface such as prophylaxis smoothing with pumice and enamel etching results in considerable damages to the enamel surface of the tooth. In this study, we have proposed optical coherence tomography as a noninvasive imaging technique for the evaluation of damage induced during such treatment procedures. Using depth intensity analysis of the obtained cross-sectional images, the damage resulting to the enamel surface was studied after prophylaxis smoothening and etching steps.

3-Dimensional characterization of cortical bone microdamage following placement of orthodontic microimplants using Optical Coherence Tomography.

Microimplants are being used extensively in clinical practice to achieve absolute anchorage. Success of microimplant mainly depend on its primary stability onto the cortical bone surface and the associated Microdamage of the cortical bone during insertion procedure leads to many a microimplants to fail and dislodge from the cortical bone leading to its failure. Even though, previous studies showed occurrence of microdamage in the cortical bone, they were mainly 2-dimension studies or studies that were invasive to the host. In the present study, we used a non-invasive, non-ionizing imaging technique- Optical Coherence Tomography (OCT), to image and analyze the presence of microdamage along the cortical bone surrounding the microimplant. We inserted 80 microimplants in two different methods (drill and drill free method) and in two different angulations onto the cortical bone surface. Images were obtained in both 2D and 3D imaging modes. In the images, microdamage in form of microcracks on the cortical bone surface around the bone-microimplant interface and micro-elevations of the cortical bone in angulated microimplant insertions and the presence of bone debris due to screwing motion of the microimplant on insertion can be appreciated visually and quantitatively through the depth intensity profile analysis of the images.

Assessment of cortical bone microdamage following insertion of microimplants using optical coherence tomography: a preliminary study.

OBJECTIVES: The study was done to evaluate the efficacy of optical coherence tomography (OCT), to detect and analyze the microdamage occurring around the microimplant immediately following its placement, and to compare the findings with micro-computed tomography (µCT) images of the samples to validate the result of the present study. METHODS: Microimplants were inserted into bovine bone samples. Images of the samples were obtained using OCT and µCT. Visual comparisons of the images were made to evaluate whether anatomical details and microdamage induced by microimplant insertion were accurately revealed by OCT. RESULTS: The surface of the cortical bone with its anatomical variations is visualized on the OCT images. Microdamage occurring on the surface of the cortical bone around the microimplant can be appreciated in OCT images. The resulting OCT images were compared with the µCT images. A high correlation regarding the visualization of individual microcracks was observed. The depth penetration of OCT is limited when compared to µCT. CONCLUSIONS: OCT in the present study was able to generate high-resolution images of the microdamage occurring around the microimplant. Image quality at the surface of the cortical bone is above par when compared with µCT imaging, because of the inherent high contrast and high-resolution quality of OCT systems. Improvements in the imaging depth and development of intraoral sensors are vital for developing a real-time imaging system and integrating the system into orthodontic practice.

Biophotonic approach for the characterization of initial bitter-rot progression on apple specimens using optical coherence tomography assessments.

The tremendous advances achieved in the biophotonics technologies have intensified the necessity for non-invasive modalities that can characterize diverse biological materials with increased sensitivity and resolution. Optical coherence tomography (OCT) is one of the techniques that has been applied for biological applications in medicine and agriculture to identify structural properties. Herein, we report the successful incorporation of OCT for the identification of morphological changes that occur as a result of the bitter rot disease, through continuous detection of structural changes. Detailed inner morphological structural changes occurring in fruit specimens were precisely analyzed as a function of the disease incubation period using OCT. The conducted histological correlation and quantitative three-dimensional evaluations provide a robust platform for further discoveries related to plant materials. The results highlight the initial identification of bitter rot progression on apple specimens owing to the non-invasive inspection capability of OCT. Therefore, we expect that the proposed method will enable immediate sensitivity improvements in the inspection of plant diseases for postharvest utility.

Non-Destructive Classification of Diversely Stained Capsicum annuum Seed Specimens of Different Cultivars Using Near-Infrared Imaging Based Optical Intensity Detection.

The non-destructive classification of plant materials using optical inspection techniques has been gaining much recent attention in the field of agriculture research. Among them, a near-infrared (NIR) imaging method called optical coherence tomography (OCT) has become a well-known agricultural inspection tool since the last decade. Here we investigated the non-destructive identification capability of OCT to classify diversely stained (with various staining agents) Capsicum annuum seed specimens of different cultivars. A swept source (SS-OCT) system with a spectral band of 1310 nm was used to image unstained control C. annuum seeds along with diversely stained Capsicum seeds, belonging to different cultivar varieties, such as C. annuum cv. PR Ppareum, C. annuum cv. PR Yeol, and C. annuum cv. Asia Jeombo. The obtained cross-sectional images were further analyzed for the changes in the intensity of back-scattered light (resulting due to dye pigment material and internal morphological variations) using a depth scan profiling technique to identify the difference among each seed category. The graphically acquired depth scan profiling results revealed that the control specimens exhibit less back-scattered light intensity in depth scan profiles when compared to the stained seed specimens. Furthermore, a significant back-scattered light intensity difference among each different cultivar group can be identified as well. Thus, the potential capability of OCT based depth scan profiling technique for non-destructive classification of diversely stained C. annum seed specimens of different cultivars can be sufficiently confirmed through the proposed scheme. Hence, when compared to conventional seed sorting techniques, OCT can offer multipurpose advantages by performing sorting of seeds in respective to the dye staining and provides internal structural images non-destructively.

In Vivo Non-Destructive Monitoring of Capsicum Annuum Seed Growth with Diverse NaCl Concentrations Using Optical Detection Technique.

We demonstrate that optical coherence tomography (OCT) is a plausible optical tool for in vivo detection of plant seeds and its morphological changes during growth. To investigate the direct impact of salt stress on seed germination, the experiment was conducted using Capsicum annuum seeds that were treated with different molar concentrations of NaCl. To determine the optimal concentration for the seed growth, the seeds were monitored for nine consecutive days. In vivo two-dimensional OCT images of the treated seeds were obtained and compared with the images of seeds that were grown using sterile distilled water. The obtained results confirm the feasibility of using OCT for the proposed application. Normalized depth profile analysis was utilized to support the conclusions.

Non-Destructive Analysis of the Internal Anatomical Structures of Mosquito Specimens Using Optical Coherence Tomography.

The study of mosquitoes and analysis of their behavior are of crucial importance in the on-going efforts to control the alarming increase in mosquito-borne diseases. Furthermore, a non-destructive and real-time imaging technique to study the anatomical features of mosquito specimens can greatly aid the study of mosquitoes. In this study, we demonstrate the three-dimensional imaging capabilities of optical coherence tomography (OCT) for structural analysis of Anopheles sinensis mosquitoes. The anatomical features of An. sinensis head, thorax, and abdominal regions, along with the morphology of internal structures, such as foregut, midgut, and hindgut, were studied using OCT imaging. Two-dimensional and three-dimensional OCT images, used in conjunction with histological images, proved useful for anatomical analysis of mosquito specimens. By presenting this work as an initial study, we demonstrate the applicability of OCT for future mosquito-related entomological research, and also in identifying changes in mosquito anatomical structure.

Optical coherence tomography-integrated, wearable (backpack-type), compact diagnostic imaging modality for in situ leaf quality assessment: publisher's note.

This note points out additional funding that was not added to Appl. Opt.56, D108 (2017)APOPAI0003-693510.1364/AO.56.00D108.

Optical coherence tomography-integrated, wearable (backpack-type), compact diagnostic imaging modality for in situ leaf quality assessment.

We developed a compact, wearable diagnostic imaging modality employing optical coherence tomography for in situ plant leaf quality assessments. This system is capable of diagnosing infected leaves at the initial disease stages. Our system is a versatile backpack-type imaging modality with a compact spectrometer, miniature computer, rechargeable power source, and handheld inspection probe. This method enhances real-time in situ specimen inspection through direct implementation of the imaging modality in a plantation. To evaluate the initial performance, field experiments were conducted in apple, pear, and persimmon plantations. Based on the obtained results, we can conclude that the developed imaging modality can be considered as a promising, efficient, convenient, and fast in situ inspection technique for various agricultural fields, which minimizes the limitations of complex tabletop inspection modalities.

Depth enhancement in spectral domain optical coherence tomography using bidirectional imaging modality with a single spectrometer.

A method for depth enhancement is presented using a bidirectional imaging modality for spectral domain optical coherence tomography (SD-OCT). Two precisely aligned sample arms along with two reference arms were utilized in the optical configuration to scan the samples. Using exemplary images of the optical resolution target, Scotch tape, a silicon sheet with two needles, and a leaf, we demonstrated how the developed bidirectional SD-OCT imaging method increases the ability to characterize depth-enhanced images. The results of the developed system were validated by comparing the images with the standard OCT configuration (single-sample arm setup). Given the advantages of higher resolution and the ability to visualize deep morphological structures, this method can be utilized to increase the depth dependent fall-off in samples with limited thickness. Thus, the proposed bidirectional imaging modality is apt for cross-sectional imaging of entire samples, which has the potential capability to improve the diagnostic ability.