Human middle-ear muscle pulls change tympanic-membrane shape and low-frequency middle-ear transmission magnitudes and delays.

The three-bone flexible ossicular chain in mammals may allow independent alterations of middle-ear (ME) sound transmission via its two attached muscles, for both acoustic and non-acoustic stimuli. The tensor tympani (TT) muscle, which has its insertion on the malleus neck, is thought to increase tension of the tympanic membrane (TM). The stapedius (St) muscle, which has its insertion on the stapes posterior crus, is known to stiffen the stapes annular ligament. We produced ME changes in human cadaveric temporal bones by statically pulling on the TT and St muscles. The 3D static TM shape and sound-induced umbo motions from 20 Hz to 10 kHz were measured with optical coherence tomography (OCT); stapes motion was measured using laser-Doppler vibrometry (LDV). TT pulls made the TM shape more conical and moved the umbo medially, while St pulls moved the umbo laterally. In response to sound below about 1 kHz, stapes-velocity magnitudes generally decreased by about 10 dB due to TT pulls and 5 dB due to St pulls. In the 250 to 500 Hz region, the group delay calculated from stapes-velocity phase showed a decrease in transmission delay of about 150 µs by TT pulls and 60 µs by St pulls. Our interpretation of these results is that ME-muscle activity may provide a way of mechanically changing interaural time- and level-difference cues. These effects could help the brain align head-centered auditory and ocular-centered visual representations of the environment.

Cochlear motion across the reticular lamina implies that it is not a stiff plate.

Within the cochlea, the basilar membrane (BM) is coupled to the reticular lamina (RL) through three rows of piezo-like outer hair cells (OHCs) and supporting cells that endow mammals with sensitive hearing. Anatomical differences across OHC rows suggest differences in their motion. Using optical coherence tomography, we measured in vivo and postmortem displacements through the gerbil round-window membrane from approximately the 40-47 kHz best-frequency (BF) regions. Our high spatial resolution allowed measurements across the RL surface at the tops of the three rows of individual OHCs and their bottoms, and across the BM. RL motion varied radially; the third-row gain was more than 3 times greater than that of the first row near BF, whereas the OHC-bottom motions remained similar. This implies that the RL mosaic, comprised of OHC and phalangeal-process tops joined together by adhesion molecules, is much more flexible than the Deiters' cells connected to the OHCs at their bottom surfaces. Postmortem, the measured points moved together approximately in phase. These imply that in vivo, the RL does not move as a stiff plate hinging around the pillar-cell heads near the first row as has been assumed, but that its mosaic-like structure may instead bend and/or stretch.

Cochlear Fluid Spaces and Structures of the Gerbil High-Frequency Region Measured Using Optical Coherence Tomography (OCT).

Since it has been difficult to directly observe the morphology of the living cochlea, our ability to infer the mechanical functioning of the living ear has been limited. Nearly all our knowledge about cochlear morphology comes from postmortem tissue that was fixed and processed using procedures that possibly distort the structures and fluid spaces of the organ of Corti. In this study, optical coherence tomography was employed to obtain volumetric images of the high-frequency hook region of the gerbil cochlea, as viewed through the round window, with far better resolution capability than had been possible before. The anatomical structures and fluid spaces of the organ of Corti were segmented and quantified in vivo and over a 90-min postmortem period. We find that the arcuate-zone and pectinate-zone widths change very little postmortem. The volume of the scala tympani between the round-window membrane and basilar membrane and the volume of the inner spiral sulcus decrease in the first 60-min postmortem. While textbook drawings of the mammalian organ of Corti and cortilymph prominently depict the tunnel of Corti, the outer tunnel is typically missing. This is likely because textbook drawings are typically made from images obtained by histological methods. Here, we show that the outer tunnel is nearly twice as big as the tunnel of Corti or the space of Nuel. This larger outer tunnel fluid space could have a substantial, little-appreciated effect on cochlear micromechanics. We speculate that the outer tunnel forms a resonant structure that may affect reticular-lamina motion.

Label-free optical projection tomography for quantitative three-dimensional anatomy of mouse embryo.

Recent progress in three-dimensional optical imaging techniques allows visualization of many comprehensive biological specimens. Optical clearing methods provide volumetric and quantitative information by overcoming the limited depth of light due to scattering. However, current imaging technologies mostly rely on the synthetic or genetic fluorescent labels, thus limits its application to whole-body visualization of generic mouse models. Here, we report a label-free optical projection tomography (LF-OPT) technique for quantitative whole mouse embryo imaging. LF-OPT is based on the attenuation contrast of light rather than fluorescence, and it utilizes projection imaging technique similar to computed tomography for visualizing the volumetric structure. We demonstrate this with a collection of mouse embryo morphologies in different stages using LF-OPT. Additionally, we extract quantitative organ information applicable toward high-throughput phenotype screening. Our results indicate that LF-OPT can provide multi-scale morphological information in various tissues including bone, which can be difficult in conventional optical imaging technique.

In Vivo Vibration Measurement of Middle Ear Structure Using Doppler Optical Coherence Tomography: Preliminary Study.

OBJECTIVES: Doppler optical coherence tomography (DOCT) is useful for both, the spatially resolved measurement of the tympanic membrane (TM) oscillation and high-resolution imaging. We demonstrated a new technique capable of providing real-time two-dimensional Doppler OCT image of rapidly oscillatory latex mini-drum and in vivo rat TM and ossicles. METHODS: Using DOCT system, the oscillation of sample was measured at frequency range of 1-4 kHz at an output of 15 W. After the sensitivity of the DOCT system was verified using a latex mini-drum consisting of a 100 µm-thick latex membrane, changes in displacement of the umbo and contacted area between TM and malleus in normal and pathologic conditions. RESULTS: The oscillation cycles of the mini-drum for stimulus frequencies were 1.006 kHz for 1 kHz, 2.012 kHz for 2kHz, and 3.912 kHz for 4 kHz, which means that the oscillation cycle of the mini-drum become short in proportional to the frequency of stimuli. The oscillation cycles of umbo area and the junction area in normal TM for frequencies of the stimuli showed similar integer ratio with the data of latex mini-drum for stimuli less than 4 kHz. In the case of middle ear effusion condition, the Doppler signal showed a tendency of attenuation in all frequencies, which was prominent at 1 kHz and 2 kHz. CONCLUSION: The TM vibration under sound stimulation with frequencies from 1 kHz to 4 kHz in normal and pathologic conditions was demonstrated using signal demodulation method in in vivo condition. The OCT technology could be helpful for functional and structural assessment as an optional modality.

Optical assessment of the in vivo tympanic membrane status using a handheld optical coherence tomography-based otoscope.

OBJECTIVE: Conventional otoscopes and oto-endoscopes, which are used to examine the tympanic membrane (TM), do not provide tomographic information. Optical coherence tomography (OCT) non-invasively reveals the depth-resolved internal microstructure of the TM with very high spatial resolution. We designed this study to examine the TMs with middle ear diseases using a handheld otoscope employing 860 nm spectral domain (SD)-OCT, combined with video camera and to demonstrate the clinical applicability of this system. DESIGN: A total of 120 patients with otologic symptoms were enrolled. TM images were obtained using the handheld OCT-based otoscope (860 nm central wave length, 15 µm axial resolution, 15 µm lateral resolution, and 7 mm scanning range using relay lens). Both OCT and oto-endoscope images were compared according to the clinical characteristics such as perforation, retraction, and postoperative healing process. RESULTS: The objective grade about the thickness of perforation margins and the accurate information about the extent of TM retraction that was not distinguishable by oto-endoscopic exam could be identified using this system. The postoperative healing process of TMs could be also followed using the OCT device. CONCLUSION: These analyses from the surgeon-oriented perspective suggest another useful application of the handheld OCT device.

In vivo 3D imaging of the human tympanic membrane using a wide-field diagonal-scanning optical coherence tomography probe.

A wide-field optical coherence tomography (OCT) probe was developed that adapts a diagonal-scanning scheme for three-dimensional (3D) in vivo imaging of the human tympanic membrane. The probe consists of a relay lens to enhance the lateral scanning range up to 7 mm. Motion artifacts that occur with the use of handheld probes were found to be decreased owing to the diagonal-scanning pattern, which crosses the center of the sample to facilitate entire 3D scans. 3D images could be constructed from a small number of two-dimensional OCT images acquired using the diagonal-scanning technique. To demonstrate the usefulness and performance of the developed system with the handheld probe, in vivo tympanic membranes of humans and animals were imaged in real time.

Bio-Photonic Detection and Quantitative Evaluation Method for the Progression of Dental Caries Using Optical Frequency-Domain Imaging Method.

The initial detection of dental caries is an essential biomedical requirement to barricade the progression of caries and tooth demineralization. The objective of this study is to introduce an optical frequency-domain imaging technique based quantitative evaluation method to calculate the volume and thickness of enamel residual, and a quantification method was developed to evaluate the total intensity fluctuation in depth direction owing to carious lesions, which can be favorable to identify the progression of dental caries in advance. The cross-sectional images of the ex vivo tooth samples were acquired using 1.3 µm spectral domain optical coherence tomography system (SD-OCT). Moreover, the advantages of the proposed method over the conventional dental inspection methods were compared to highlight the potential capability of OCT. As a consequence, the threshold parameters obtained through the developed method can be used as an efficient investigating technique for the initial detection of demineralization.

Lateral resolution enhancement using programmable phase modulator in optical coherence tomography.

Adaptive optics plays an important role in the correction of high-order aberrations to enhance lateral resolution. An OCT system coupled with a programmable phase modulator is designed to verify lateral resolution improvement. The Hamamatsu high-resolution, non-pixelized, optically addressed light modulator (PAL-SLM) PPM X7550 series is used to correct aberrations, utilizing a simple method based on phase information from OCT images. The advantages of this method are its high simplicity and low cost. A raster scanning technique is adopted to scan samples covered with scattering suspension. Metal lines covered with scattering suspension are imaged, and a respective correction is applied to remove the aberration. Results demonstrate an improvement in lateral resolution on metal grid samples.

Evaluation of the usefulness of three-dimensional optical coherence tomography in a guinea pig model of endolymphatic hydrops induced by surgical obliteration of the endolymphatic duct.

Optical coherence tomography (OCT) has advanced significantly over the past two decades and is currently used extensively to monitor the internal structures of organs, particularly in ophthalmology and dermatology. We used ethylenediamine tetra-acetic acid (EDTA) to decalcify the bony walls of the cochlea and investigated the inner structures by deep penetration of light into the cochlear tissue using OCT on a guinea pig model of endolymphatic hydrops (EH), induced by surgical obliteration of the endolymphatic duct. The structural and functional changes associated with EH were identified using OCT and auditory brainstem response tests, respectively. We also evaluated structural alterations in the cochlea using three-dimensional reconstruction of the OCT images, which clearly showed physical changes in the cochlear structures. Furthermore, we found significant anatomical variations in the EH model and conducted graphical analysis by strial atrophy for comparison. The physical changes included damage to and flattening of the organ of Corti­evidence of Reissner's membrane distention­and thinning of the lateral wall. These results indicate that observation of EDTA-decalcified cochlea using OCT is significant in examination of gradual changes in the cochlear structures that are otherwise not depicted by hematoxylin and eosin staining.

Optical coherence tomography for the diagnosis and evaluation of human otitis media.

We report the application of optical coherence tomography (OCT) to the diagnosis and evaluation of otitis media (OM). Whereas conventional diagnostic modalities for OM, including standard and pneumatic otoscopy, are limited to visualizing the surface of the tympanic membrane (TM), OCT effectively reveals the depth-resolved microstructure below the TM with very high spatial resolution, with the potential advantage of its use for diagnosing different types of OM. We examined the use of 840-nm spectral domain-OCT (SD-OCT) clinically, using normal ears and ears with the adhesive and effusion types of OM. Specific features were identified in two-dimensional OCT images of abnormal TMs, compared to images of healthy TMs. Analysis of the A-scan (axial depth scan) identified unique patterns of constituents within the effusions. The OCT images could not only be used to construct a database for the diagnosis and classification of OM but OCT might also represent an upgrade over current otoscopy techniques.

In vivo imaging of middle-ear and inner-ear microstructures of a mouse guided by SD-OCT combined with a surgical microscope.

We developed an augmented-reality system that combines optical coherence tomography (OCT) with a surgical microscope. By sharing the common optical path in the microscope and OCT, we could simultaneously acquire OCT and microscope views. The system was tested to identify the middle-ear and inner-ear microstructures of a mouse. Considering the probability of clinical application including otorhinolaryngology, diseases such as middle-ear effusion were visualized using in vivo mouse and OCT images simultaneously acquired through the eyepiece of the surgical microscope during surgical manipulation using the proposed system. This system is expected to realize a new practical area of OCT application.

Development of real-time dual-display handheld and bench-top hybrid-mode SD-OCTs.

Development of a dual-display handheld optical coherence tomography (OCT) system for retina and optic-nerve-head diagnosis beyond the volunteer motion constraints is reported. The developed system is portable and easily movable, containing the compact portable OCT system that includes the handheld probe and computer. Eye posterior chambers were diagnosed using the handheld probe, and the probe could be fixed to the bench-top cradle depending on the volunteers' physical condition. The images obtained using this handheld probe were displayed in real time on the computer monitor and on a small secondary built-in monitor; the displayed images were saved using the handheld probe's built-in button. Large-scale signal-processing procedures such as k-domain linearization, fast Fourier transform (FFT), and log-scaling signal processing can be rapidly applied using graphics-processing-unit (GPU) accelerated processing rather than central-processing-unit (CPU) processing. The Labview-based system resolution is 1,024 × 512 pixels, and the frame rate is 56 frames/s, useful for real-time display. The 3D images of the posterior chambers including the retina, optic-nerve head, blood vessels, and optic nerve were composed using real-time displayed images with 500 × 500 × 500 pixel resolution. A handheld and bench-top hybrid mode with a dual-display handheld OCT was developed to overcome the drawbacks of the conventional method.

Wavelength-Filter Based Spectral Calibrated Wave number - Linearization in 1.3 mm Spectral Domain Optical Coherence.

In this study, we demonstrate the enhanced spectral calibration method for 1.3 µm spectral-domain optical coherence tomography (SD-OCT). The calibration method using wavelength-filter simplifies the SD-OCT system, and also the axial resolution and the entire speed of the OCT system can be dramatically improved as well. An externally connected wavelength-filter is utilized to obtain the information of the wavenumber and the pixel position. During the calibration process the wavelength-filter is placed after a broadband source by connecting through an optical circulator. The filtered spectrum with a narrow line width of 0.5 nm is detected by using a line-scan camera. The method does not require a filter or a software recalibration algorithm for imaging as it simply resamples the OCT signal from the detector array without employing rescaling or interpolation methods. One of the main drawbacks of SD-OCT is the broadened point spread functions (PSFs) with increasing imaging depth can be compensated by increasing the wavenumber-linearization order. The sensitivity of our system was measured at 99.8 dB at an imaging depth of 2.1 mm compared with the uncompensated case.

Non-destructive inspection methods for LEDs using real-time displaying Optical Coherence Tomography.

In this study, we report the applicability of two different Optical Coherence Tomography (OCT) technologies for inspecting Light Emitting Diode (LED) structures. Sectional images of a LED were captured using a Spectral Domain OCT (SD-OCT) system and a Swept Source OCT (SS-OCT) system. Their center wavelengths are 850 and 1,310 nm, respectively. We acquired cross-sectional two dimensional (2D) images of a normal LED and extracted sectional profiles to inspect possible wire disconnection that may be present in the LED manufacturing process. The SD-OCT and SS-OCT images were compared with each other in the same sample to study their advantages. The distribution of fluorescence material was observed more clearly with the SD-OCT of 850 nm wavelength, whereas the status of wire connection was clearer in the SS-OCT images with 1,310 nm wavelength. In addition, the volume of the fluorophore space was calculated from the OCT images. This is the first report that a nondestructive optical imaging modality such as OCT can be applied to finding screen defects in LED. We expect this method can improve the inspection efficacy over traditional inspection methods such as Charged Coupled Device (CCD) camera or X-ray instruments.

Ultra-fast displaying Spectral Domain Optical Doppler Tomography system using a Graphics Processing Unit.

We demonstrate an ultrafast displaying Spectral Domain Optical Doppler Tomography system using Graphics Processing Unit (GPU) computing. The calculation of FFT and the Doppler frequency shift is accelerated by the GPU. Our system can display processed OCT and ODT images simultaneously in real time at 120 fps for 1,024 pixels × 512 lateral A-scans. The computing time for the Doppler information was dependent on the size of the moving average window, but with a window size of 32 pixels the ODT computation time is only 8.3 ms, which is comparable to the data acquisition time. Also the phase noise decreases significantly with the window size. Since the performance of a real-time display for OCT/ODT is very important for clinical applications that need immediate diagnosis for screening or biopsy. Intraoperative surgery can take much benefit from the real-time display flow rate information from the technology. Moreover, the GPU is an attractive tool for clinical and commercial systems for functional OCT features as well.