We present an advanced technique of polarized light microscopy (PLM) for the assessment of carious lesions, wherein the degree of polarization (DOP) is evaluated. Through modifications of a conventional PLM setup, images were generated by calculating the DOP from an image series of different linear polarization images acquired with a polarization camera. Demineralization is reliably displayed by the DOP in accordance with the common imaging methods. Evaluating the DOP by PLM allows the characterization of the different pathohistological zones of caries with a new quantitative approach.
Polarized light microscopy (PLM) is an established technique for the inspection of thin tooth sections in dental research. However, conventional PLM is mainly based on the qualitative evaluation of color-dependent birefringence, depolarization and transmittance but lacks a quantitative interpretation and, thus, the comparison with other polarimetric imaging methods. Here, we describe an easy to implement extension of PLM that enables measuring the degree of polarization (DOP). By replacing the analyzer and RGB camera in PLM with a monochrome polarization camera, linear polarization states can be directly determined from the transmitted light. Additionally rotating the polarizer in the illumination path and, by that, the state incident into the sample facilitates reconstructing a DOP image from multiple linear measurements. The resulting depolarization measurements are compared with conventional PLM images as well as X-ray micro-computed tomography (μCT) data of the intact teeth. Our results show that caries and demineralization appear to be directly related to depolarization in enamel. However, the interpretation is more complicated for dentin, which shows a reduced DOP also in sound tissue. We assume that these insights support the development and analysis of future dental polarimetry techniques in vivo, such as intra-oral polarization-sensitive optical coherence tomography (PS-OCT)
Caries of the interdental space are usually difficult to detect with conventional bitewing radiography. We have developed an interdental probe for polarization-sensitive optical coherence tomography (PSOCT) that can distinguish between healthy and carious tooth structures. The development is based on a fiber optic probe with a compact motor design that allows volumetric imaging. In a clinical study, demineralization was detected using proximal PSOCT scans of premolars and molars, and compared with blind diagnoses from four dentists. The results show that PSOCT can detect both, early demineralization and advanced proximal caries, some of which could not be diagnosed with bitewing radiography.
Modern noninvasive imaging techniques are used in many areas of medical research, particularly where an additional modality is required to support close monitoring during pre- or post-care. In oral medicine, the combination of an experienced clinician and supporting noninvasive optical devices for oral imaging, such as optical coherence tomography (OCT), may ensure improved diagnosis. In this study, the pathological features of lichenoid oral mucosa were noninvasively imaged with our miniaturized pen-like OCT probe and compared to adjacent healthy areas. The non-homogeneous epithelial (EP) layer, increased cornification of the EP surface and unidentifiable basement membrane are the most important features recorded.
Common-path probes provide considerable advantages for fiber-based OCT due to intrinsic length and phase matching. However, the polarization state of the reference light is usually arbitrary and variable due to stress-induced birefringence in single-mode fibers, which complicates implementing polarization-sensitive OCT. Here, we present depth-resolved retardation measurements with a single-mode fiber-based common-path probe by utilizing the constrained polarization evolution and the mirror state phenomenon for reconstruction of the round-trip measurements in the case of arbitrary reference states. Thus, a compact and flexible polarization-sensitive OCT implementation is demonstrated.
We present a miniaturized handheld OCT probe (9 grams weight), approximately the size of a small pen (10 mm x 140 mm), developed for use inside a patient’s mouth for examination of the oral mucosa. The probe operates in common-path mode and uses a magnetic scanning system to actuate a lensed fibre, achieving 50 B-scans per second. The system is demonstrated with OCT imaging of the buccal and alveolar mucosa of six patients with oral lichen planus (OLP) during clinical routine examination, and showed pathological changes in the tissue microarchitecture.
Concerning the usage of non-invasive optical techniques for oral cancer detection, the presented study dealt with the manual and semi-automatic measurement of oral epithelial thickness by means of optical coherence tomography (OCT). Here, a total of 3,510 OCT images out of seven areas of the oral cavity was analyzed. In the result, epithelial thickness varied from 77 µm to 704 µm. Beyond that, there was a good correlation between both methods (ρ = 0.71-0.96). With regard to clinical routine use, manual epithelial thickness measurement could be beneficial due to a currently lower time investment with comparable precision.
Optical coherence tomography (OCT) is a non-invasive, radiation-free optical technique for caries diagnosis. The presented study dealt with the correlation of conventional caries detection methods and using polarisation-sensitive OCT (PS-OCT). Human extracted carious teeth were assessed by dentists with visual and radiographic examination and PS-OCT images were gained. Proximal lesions could be detected early in the intensity-based OCT cross-sectional images. The degree of polarization can clearly delineate proximal caries from surrounding healthy tooth structure. The obtained results pave the way to reduce invasive filling therapy through early possibilities of intervention.
Depth-resolved retardation measurements with a single-mode fiber-based common-path probe by using polarization-sensitive optical coherence tomography are presented, utilizing the constrained polarization evolution and the mirror state phenomenon for reconstruction of the round-trip measurements.
To date, there is still a lack of reliable imaging modalities to improve the quality of medical examinations of the oral mucosa in dentistry. One promising imaging technologies is optical coherence tomography (OCT). The present study provides in vivo intensity-based OCT cross sections of the human oral mucosa of the anterior and posterior oral cavity as well as the oropharynx of 47 disease-free adult volunteers. The randomized blinded image analysis was performed by means of 3,560 OCT cross sections using a set of seven qualitative and quantitative criteria to assess the epithelium and the lamina propria.
The prerequisite for a successful caries management is the early and precise detection of lesions. We have recently shown that depolarization imaging based on polarization-sensitive optical coherence tomography (PS-OCT) is a promising tool to detect demineralized enamel. To validate this contrast mechanism, we acquired data sets of tooth samples with occlusal caries lesions using PS-OCT and x-ray micro-computed tomography (µCT). Spatial alignment of the PS-OCT and µCT volumes was achieved by creating point clouds from surface segmentations, and subsequent registration. PS-OCT depolarization signals are in good agreement with the µCT results, showing the potential of this approach for caries diagnostics.
Optical coherence tomography (OCT) is a contactless and non-invasive imaging technique. Due to the high resolution of some 10 μm and the penetration depth of 1-2 mm in scattering tissue, OCT closes the gap between microscopy and sonography. Here, we present a GRIN-based endoscopic OCT (eOCT) system for middle ear diagnostics. EOCT combines the benefit of endoscopic imaging and the advantage of morphological and functional investigation of the tympanic membrane (TM). The eOCT system has a working distance of 10 mm, which results in a field-of-view of 10 mm. This allows a full three-dimensional visualization of the TM and surrounding tissue. In addition, the oscillation of the TM can be measured spatially resolved and in the frequency range between 500 Hz and 5000 Hz with 125 Hz resolution, which is realized by phase-resolved Doppler-OCT. First clinical results are demonstrated for one selected cases, a middle ear effusion.
While dental caries represents the major chronic disease of humans, visual and tactile inspection are the methods of choice in daily dental practice. Several optical technologies have been developed in recent years for the purpose of dental examination, including near-infrared light transillumination as a promising tool for the supplementation and partial replacement of radiography. In case of occlusal alterations, the incidence of surface discolorations impedes the visual assessment, whereas tactile inspection appears to yield little diagnostic information or might be detrimental. Optical coherence tomography (OCT) facilitates depth-resolved imaging with μm resolution, utilizing near-infrared light, and has already shown its potential for various dental applications. We have recently demonstrated that depolarization imaging utilizing the degree of polarization uniformity (DOPU) extends those abilities by the detection of early carious lesions, as it provides an unambiguous demineralization contrast. Here, we show that this approach also enables the assessment of occlusal enamel lesions in the presence of stains, and compare PS-OCT cross sections with polarized light microscopy (PLM) images of thin sections. For tooth samples with discoloration or demineralization, respectively, PS-OCT and PLM results are in good agreement.
The detection of incipient carious lesions is crucial for the preservation of dental hard tissue with preventive therapies. Optical coherence tomography (OCT) based imaging has been proofed to be a valuable methodology for the depth-resolved detection of alterations at dental hard tissue. Due to changes of birefringent properties at carious lesions, the measurement of polarization properties with polarization-sensitive OCT (PS-OCT) provides an additional contrast that could enhance the detection of incipient carious lesions. The aim of the present study was to monitor changes of polarization properties during the initial demineralization process by means of PS-OCT. Therefore, 18 bovine enamel palates were demineralized up to 7 weeks in an artificial demineralization model using lactate acid and buffer solution to mimic natural caries progression. The palates were stored at pH 5.5 and 35°C with constant fluid movement. In order to detect changes of depolarization at different stages of demineralization, the degree of polarization uniformity (DOPU) was calculated from PS-OCT measurements, performed under wet and dry conditions. In addition, polarization microscopy and micro-computed tomography of thin sections were analyzed to assess lesion depth and mineral loss. Our main finding is that consecutive stages of demineralization are correlated to an increasing DOPU contrast between sound enamel and the lesion area, where the lesion area is connected to decreasing DOPU values. Our results show that PS-OCT based imaging of birefringence changes is sensitive to very early stages of the demineralization process and adds a valuable contrast for the detection and monitoring of incipient enamel lesions.
For future non-invasive optical biopsy by OCT, there is a demand to relate the obtained scans with histological cross sections, which represent the diagnostic gold standard. For this reason, the aim of the performed study was the qualitative matching of OCT cross-sectional images with histological pictures from various image databases. Therefore, the human oral mucosa of forty-seven healthy volunteers was investigated at ten different regions, e.g. lip, floor of mouth, hard and soft palate, by using the recently proposed in vivo endoscopic rigid handheld OCT system (central wavelength: λ = 840 nm, spatial resolution: 11.6 μm axial and 17.4 μm lateral). The results allow answering the question, whether OCT imaging is able to detect sublayers of mucosa and to clearly identify tissue structures by using a qualitative scoring system. In fact, depiction of epithelium, lamina propria and configurations like blood vessels and salivary glands, respectively, is possible, whereas a sufficient visualization of deep submucosa or epithelial sublayers cannot be achieved with the used wavelength range. Despite a well-defined surface profile, evaluating the level of keratinization is challenging. In contrast to histological cross sections impaired by fixation, orientation and organisation of collagen fiber bundles are highlighted better in OCT cross sections due to the in vivo situation. With regard to future clinical studies, correct interpretation of tissue structure in healthy individuals forms the basis to identify and assess pathomorphological changes related to various kinds of oral diseases by means of OCT.
An endoscopic optical coherence tomography (OCT) system with a wide field-of-view of 8 mm is presented, which combines the image capability of endoscopic imaging at the middle ear with the advantages of functional OCT imaging, allowing a morphological and functional assessment of the human tympanic membrane. The endoscopic tube has a diameter of 3.5 mm and contains gradient-index optics for simultaneous forward-viewing OCT and video endoscopy. The endoscope allows the three-dimensional visualization of nearly the entire tympanic membrane. In addition, the oscillation of the tympanic membrane is measured spatially resolved and in the frequency range between 500 Hz and 5 kHz with 125 Hz resolution, which is realized by phase-resolved Doppler OCT imaging during acoustical excitation with chirp signals. The applicability of the OCT system is demonstrated in vivo. Due to the fast image acquisition, structural and functional measurements are only slightly affected by motion artifacts.
The common way to diagnose hard and soft tissue irregularities in the oral cavity is initially the visual inspection by an experienced dentist followed by further medical examinations, such as radiological imaging and/or histopathological investigation. For the diagnosis of oral hard and soft tissues, the detection of early transformations is mostly hampered by poor visual access, low specificity of the diagnosis techniques, and/or limited feasibility of frequent screenings. Therefore, optical noninvasive diagnosis of oral tissue is promising to improve the accuracy of oral screening. Considering this demand, a rigid handheld endoscopic scanner was developed for optical coherence tomography (OCT). The novelty is the usage of a commercially near-infrared endoscope with fitting optics in combination with an established spectral-domain OCT system of our workgroup. By reaching a high spatial resolution, in vivo images of anterior and especially posterior dental and mucosal tissues were obtained from the oral cavity of two volunteers. The convincing image quality of the endoscopic OCT device is particularly obvious for the imaging of different regions of the human soft palate with highly scattering fibrous layer and capillary network within the lamina propria.
As dental caries is one of the most common diseases, the early and noninvasive detection of carious lesions plays an important role in public health care. Optical coherence tomography (OCT) with its ability of depth-resolved, high-resolution, noninvasive, fast imaging has been previously recognized as a promising tool in dentistry. Additionally, polarization sensitive imaging provides quantitative measures on the birefringent tissue properties and can be utilized for imaging dental tissue, especially enamel and dentin. By imaging three exemplary tooth samples ex vivo with proximal white spot, brown spot, and cavity, we show that the combination of polarization sensitive OCT and the degree of polarization uniformity (DOPU) algorithm is a promising approach for the detection of proximal carious lesions due to the depolarization contrast of demineralized tissue. Furthermore, we investigate different sizes of the DOPU evaluation kernel on the resulting contrast and conclude a suitable value for this application. We propose that DOPU provides an easy to interpret image representation and appropriate contrast for possible future screening applications in early caries diagnostics.
Since optical coherence tomography (OCT) provides three-dimensional high-resolution images of biological tissue, the benefit of polarization contrast in the field of dentistry is highlighted in this study. Polarization-sensitive OCT (PS OCT) with phase-sensitive recording is used for imaging dental and mucosal tissues in the human oral cavity in vivo. An enhanced polarization contrast of oral structures is reached by analyzing the signals of the co- and crosspolarized channels of the swept source PS OCT system quantitatively with respect to reflectivity, retardation, optic axis orientation, and depolarization. The calculation of these polarization parameters enables a high tissue-specific contrast imaging for the detailed physical interpretation of human oral hard and soft tissues. For the proof-of-principle, imaging of composite restorations and mineralization defects at premolars as well as gingival, lingual, and labial oral mucosa was performed in vivo within the anterior oral cavity. The achieved contrast-enhanced results of the investigated human oral tissues by means of polarization-sensitive imaging are evaluated by the comparison with conventional intensity-based OCT.
In spectral domain optical coherence tomography (SD-OCT), any transverse motion component of a detected obliquely moving sample results in a nonlinear relationship between the Doppler phase shift and the axial sample velocity restricting phase-resolved Doppler OCT. To circumvent the limitation, we propose the lateral resonant Doppler flow quantification in spectral domain OCT, where the scanner movement velocity is matched to the transverse velocity component of the sample motion.
The early non-invasive diagnosis of epithelial tissue alterations in daily clinical routine is still challenging. Since optical coherence tomography (OCT) shows the potential to differentiate between benign and malignant tissue of primal endothelium, OCT could be beneficial for the early diagnosis of malignancies in routine health checks. In this research, a new handheld endoscopic scanning unit was designed and connected to a spectral domain OCT system of our workgroup for the in vivo imaging of the human oral mucosa.
Besides structural imaging, OCT can be used to estimate axial velocities of the sample resolved in depth by Doppler processing. In Fourier domain OCT (FD-OCT), this is accomplished by measuring the phase difference (i.e. phase shift) between timely separated A-scans at the same depth. In most cases, these data are disturbed by noise caused by intrinsic noise of the OCT system, specified by the SNR, and decorrelation noise caused by the transversal movement of the optical beam relative to the sample. Since the first use of Doppler methods in OCT, many methods to reduce the phase shift noise by averaging have been presented. While all these methods use a fixed set of consecutive A-scans, the best method, exhibiting no bias and having the smallest standard deviation, was questionable. Recently, Doppler processing methods depending on the mentioned noise sources and delivering the most likely phase shift and thereby axial velocity became available. The relation of these methods to previously known methods like the Kasai estimator, maximum likelihood estimator (MLE) and joint spectral and time domain OCT (jSTdOCT) will be discussed.
Recently, optical coherence tomography (OCT) was utilized in multiple studies for structural and functional imaging of the middle ear and the tympanic membrane. Since Doppler OCT allows both, the spatially resolved measurement of the tympanic membrane oscillation and high-resolution imaging, it is regarded as a promising tool for future in vivo applications. In this study, Doppler OCT is utilized for the visualization of the tympanic membrane oscillation in temporal bones with simulated Eustachian catarrh, which was realized by generating a depression in the tympanic cavity. The transfer function, meaning the oscillation amplitude normalized to the applied sound pressure, is measured frequency resolved in the range from 0.5 kHz to 6 kHz and with a lateral spatial resolution of 0.4 mm. Typical oscillation patterns could be observed in case of ambient pressure in the tympanic cavity. Under depression the characteristic oscillation patterns were observed with widely congruent appearance but at higher frequencies.
Peter Cimalla, Theresa Werner, Kai Winkler, Claudia Mueller, Sebastian Wicht, Maria Gaertner, Mirko Mehner, Julia Walther, Bernd Rellinghaus, Dierk Wittig, Mike O. Karl, Marius Ader, Richard H. Funk, Edmund Koch
Cell transplantation and stem cell therapy are promising approaches for regenerative medicine and are of interest to researchers and clinicians worldwide. However, currently, no imaging technique that allows three-dimensional in vivo inspection of therapeutically administered cells in host tissues is available. Therefore, we investigate magnetomotive optical coherence tomography (MM-OCT) of cells labeled with magnetic particles as a potential noninvasive cell tracking method. We develop magnetomotive imaging of mesenchymal stem cells for future cell therapy monitoring. Cells were labeled with fluorescent iron oxide nanoparticles, embedded in tissue-mimicking agar scaffolds, and imaged using a microscope setup with an integrated MM-OCT probe. Magnetic particle-induced motion in response to a pulsed magnetic field of 0.2 T was successfully detected by OCT speckle variance analysis, and cross-sectional and volumetric OCT scans with highlighted labeled cells were obtained. In parallel, fluorescence microscopy and laser speckle reflectometry were applied as two-dimensional reference modalities to image particle distribution and magnetically induced motion inside the sample, respectively. All three optical imaging modalities were in good agreement with each other. Thus, magnetomotive imaging using iron oxide nanoparticles as cellular contrast agents is a potential technique for enhanced visualization of selected cells in OCT.
Recent studies in animal models provided proof-of-principle evidence for cell transplantation as a potential future
therapeutic approach for retinal pathologies in humans such as Retinitis pigmentosa or age-related macular degeneration.
In this case, donor cells are injected into the eye in order to protect or replace degenerating photoreceptors or retinal
pigment epithelium. However, currently there is no three-dimensional imaging technique available that allows tracking
of cell migration and integration into the host tissue under in vivo conditions. Therefore, we investigate about
magnetomotive optical coherence tomography (OCT) of substances labeled with iron oxide nanoparticles as a potential
method for noninvasive, three-dimensional cell tracking in the retina. We use a self-developed spectral domain OCT
system for high-resolution imaging in the 800 nm-wavelength region. A suitable AC magnetic field for magnetomotive
imaging was generated using two different setups, which consist of an electrically driven solenoid in combination with a
permanent magnet, and a mechanically driven all-permanent magnet configuration. In the sample region the maximum
magnetic flux density was 100 mT for both setups, with a field gradient of 9 T/m and 13 T/m for the solenoid and the allpermanent
magnet setup, respectively. Magnetomotive OCT imaging was performed in elastic tissue phantoms and
single cells labeled with iron oxide nanoparticles. Particle-induced sub-resolution movement of the elastic samples and
the single cells could successfully be detected and visualized by means of phase-resolved Doppler OCT analysis.
Therefore, this method is a potential technique to enhance image contrast of specific cells in OCT.
The unbiased complex algorithm for flow velocity measurements with enhanced joint
spectral and time domain OCT (enhSTdOCT) is verified statistically and experimentally to find
the optimal parameters for maximal velocity noise reduction.
One current challenge of studying human tympanic membranes (TM) with optical coherence tomography (OCT) is the implementation of optics that avoid direct contact with the inflamed tissue. At the moment, no commercial device is available. We report an optics design for contactless forward imaging endoscopic optical coherence tomography (EOCT) with a large working distance (WD) and a broad field of view (FOV) by restricting the overall diameter of the probe to be small (3.5 mm), ensuring a sufficient numerical aperture. Our system uses a gradient-index (GRIN) relay lens and a GRIN objective lens, and executes a fan-shaped optical scanning pattern. The WD and FOV can be adjusted by manually changing the distance between the triplet and the GRIN relay lens. The measured lateral resolution is ∼28 μm at a WD of 10 mm with a FOV of 10 mm. Additionally, a camera and an illumination beam path were implemented within the probe for image guidance during investigations of the TM. We demonstrated the performance of the EOCT design by 3-D imaging of a human TM ex vivo and in vivo with a k-linear spectral domain OCT system.
In most dual-band OCT systems, there is a spectral gap between both bands. This might be as large as one third of the
total spectral region. Therefore, a simple Fourier transformation of the data does not give the resolution that could be
possible considering the overall spectral width. Instead, the full width of the peak is comparable to the width resulting
from a single band and is additionally modulated. We compare several methods to achieve a high resolution in spite of
the missing data. Because in a dual-band system the image quality resulting from the full information is not known we
test and optimize different algorithms by using the data from a single band system and excluding an equal part of the
spectrum. While methods using non-equidistant sample points like Vandermonde and Lomb transformation work well
with small spectral gaps, they result in large image artifacts for broader gaps, which are typical for dual-band OCT
systems. Simulations show that fitting the available data with a limited set of sine and cosines functions might give good
results but for larger gaps and appropriate amount of basis-functions this method fails, too. Dividing both bands into
overlapping smaller bands and looking at the phase of short-time Fourier transformations (STFT) resulting from a single
scatterer, it becomes clear that the amplitude of all Fourier coefficients for the total band can be estimated by the change
of the phase of the STFTs in and between both bands. Therefore, we developed an algorithm of weighting the data based
on the phase distribution of the STFT data. As a single value specifying the phase distribution we choose the absolute
sum of the STFTs divided by the sum of the amplitudes of the STFTs. Because typical OCT data are not caused by
single scatterers, we adapted this algorithm with a cluster analysis to predict the appropriate amplitude expected for a full
spectrum from the phase distribution of the STFTs inside both bands and between both bands. Although the image is
noisier and fainter compared to an image from the full spectrum, the resulting image has the best resolution from all
methods investigated.
The optical inhomogeneity of flowing blood, which appears as a waisted double fan-shaped intensity pattern inside vessels in cross-sectional optical coherence tomography (OCT) images, was investigated for the first time. High resolution spectral domain OCT in the 1.3 μm wavelength region is used to assess this inhomogeneous intravascular backscattering of light in an in vivo mouse model and flow phantom measurements. Based on a predicted alignment of the red blood cells toward laminar shear flow, an angular modulation of the corresponding backscattering cross-section inside the vessels is assumed. In combination with the signal attenuation in depth by absorption and scattering, a simple model of the intravascular intensity modulation is derived. The suitability of the model is successfully demonstrated in the in vivo experiments and confirmed by the in vitro measurements. The observed effect appears in flowing blood only and shows a strong dependency on the shear rate. In conclusion, the shear-induced red blood cell alignment in conjunction with the vessel geometry is responsible for the observed intensity distribution. This inherent effect of blood imaging has to be considered in attenuation measurements performed with OCT. Furthermore, the analysis of the intravascular intensity pattern might be useful to evaluate flow characteristics.
A method for axial resolution improvement by adequate spectral data fusion of two parallel acquired disjunct
wavelength bands in the 0.8 μm and 1.3 μm region in the field of simultaneous dual-band optical coherence tomography
(OCT) is presented. The applied spectral domain dual-band OCT system is illuminated by a supercontinuum laser light
source and allows simultaneous imaging at 800 nm and 1250 nm with free-space axial resolutions better than 4.5 μm and
7 μm, respectively, over the entire depth scan range. Each wavelength band is analyzed with an individual spectrometer
at an A-scan rate of 12 kHz. To further improve axial resolution, the 1250 nm spectra are fused with the 800 nm spectra
considering the spectrometer-inherent non-linear fringe frequency course of the interference light. The phase and
amplitude of the 1250 nm spectra are matched to the 800 nm spectra by means of short time Fourier transform analysis
in order to obtain ideally continuous joint spectra. The joint spectra then undergo conventional spectral shaping, wave
number resampling, windowing and fast Fourier transformation. First results for single A-scans of a glass slide as well as
entire cross-sectional images of biological tissue yield an axial resolution improvement of 52 % compared to
conventional single band imaging at 800 nm. The obtained A-scans show a good sharpness with a side lobe suppression
of 30 dB. Additional investigations have to be employed for the full understanding of the underlying physical
background and the optimization of the applied data processing for further image quality enhancement.
Optical coherence tomography (OCT) is an imaging modality that enables micrometer-scale contactless subsurface
imaging of biological tissue. Endoscopy, as another imaging method, has the potential of imaging tubular organs and
cavities and therefore has opened up several application areas not accessible before. The combination of OCT and
endoscopy uses the advantages of both methods and consequently allows additional imaging of structures beneath
surfaces inside cavities. Currently, visual investigations on the surface of the human tympanic membrane are possible but
only with expert eyes. up to now, visual imaging of the outer ear up to the tympanic membrane can be carried out by an
otoscope, an operating microscope or an endoscope. In contrast to these devices, endoscopy has the advantage of
imaging the whole tympanic membrane with one view. The intention of this research is the development of an
endoscopic optical coherence tomography (EOCT) device for imaging the tympanic membrane depth-resolved and
structures behind it. Detection of fluids in the middle ear, which function as an indicator for otitis media, could help to
avoid the application of antibiotics. It is possible to detect a congeries of fluids with the otoscope but the ambition is to
the early detection by OCT. The developed scanner head allows imaging in working distances in the range from zero up
to 5 mm with a field of view of 2 mm. In the next step, the scanner head should be improved to increase the working
distance and the field of view.
Recently, a new method called joint spectral and time domain optical coherence tomography (STdOCT) for flow
velocity measurement in spectral domain OCT (SD OCT) was presented. This method analyzes the detected timeresolved
interference fringe spectra by using a two-dimensional fast Fourier transformation (2D FFT) to determine
directly the Doppler frequency shift instead of calculating the phase difference at each depth position of adjacent
A-scans. There, it was found that STdOCT is more robust for measurements with low signal to noise ratio than the
classic phase-resolved Doppler OCT (DOCT) making it attractive first for imaging fast flow velocities at which a strong
Doppler angle dependent signal damping occurs due to interference fringe washout and second for investigating large
blood vessels with a big diameter and a highly damped signal of blood with increasing depth due to strong scattering and
absorption in the near-infrared wavelength range. In the present study, we would like to introduce an enhanced algorithm
for STdOCT permitting a more precise flow velocity measurement in comparison to the conventional STdOCT. The new
method determines the amplitude of the broadened Doppler frequency shift by calculating the center of gravity via the
complex analytical signal as a result of the second FFT instead of detecting the maximum intensity signal. Furthermore,
the comparison with phase-resolved DOCT was done experimentally by using a flow phantom consisting of a
1% Intralipid emulsion and a 320 μm glass capillary. As a result, the enhanced STdOCT and DOCT processed data are
completely equivalent.
KEYWORDS: Optical coherence tomography, Lung, Signal to noise ratio, Mode locking, In vivo imaging, Tissues, Coherence imaging, Single mode fibers, Visualization, Biomedical optics
Optical coherence tomography (OCT) is a noninvasive imaging modality generating cross sectional and volumetric
images of translucent samples. In Fourier domain OCT (FD OCT), the depth profile is calculated by a fast Fourier
transformation of the interference spectrum, providing speed and SNR advantage and thus making FD OCT well suitable
in biomedical applications. The interference spectrum can be acquired spectrally resolved in spectral domain OCT or
time-resolved in optical frequency domain imaging (OFDI). Since OCT images still suffer from motion artifacts,
especially under in vivo conditions, increased depth scan rates are required. Therefor, the principle of Fourier domain
mode locking has been presented by R. Huber et al. circumventing the speed limitations of conventional FD OCT
systems. In FDML lasers, a long single mode fiber is inserted in the ring resonator of the laser resulting in an optical
round trip time of a few microseconds. Sweeping the wavelength synchronously by a tunable Fabry-Perot filter can
provide wavelength sweeps with repetition rates up to a few MHz used for OFDI. Imaging of subpleural lung tissue for
investigation of lung dynamics and its elastic properties is a further biomedical application demanding high-speed OCT
imaging techniques. For the first time, the visualization of subpleural alveolar structures of a rabbit lung is presented by
the use of an FDML-based OCT system enabling repetition rates of 49.5 kHz and 122.6 kHz, respectively.
In spectral domain Doppler OCT, any transverse motion component of the obliquely moving sample relative to the
incident sample beam causes a damping of the correlation between subsequent backscattering signals or even the loss of
it making a phase-resolved Doppler flow analysis difficult because of the strong mean error of the Doppler phase shift.
To circumvent this effect, a new method for resonant Doppler flow imaging and quantification in spectral domain OCT
is proposed where the scanner movement velocity is approximately matched to the transverse velocity component of the
oblique sample motion similar to a tracking shot where the camera is moved with respect to the sample. As a result, the
backscattering signals corresponding to the moving sample will be highly correlated whereas those of static sample
structures and slowly moving scatterers will be less correlated and damped depending on the scanner velocity.
Advantageously, for the exact flow velocity quantification the new Doppler relationship of phase shift and sample
velocity has not to be applied and the linear relation of the classic Doppler model can still be used. In the present work,
first results of the lateral resonant Doppler imaging are shown for an 1 percent Intralipid flow phantom study.
Spectral domain dual-band optical coherence tomography for simultaneous imaging of rodent retina in the 0.8 μm and
1.3 μm wavelength region and non-invasive monitoring of the posterior eye microstructure in the field of retinal
degeneration research is demonstrated. The system is illuminated by a supercontinuum laser source and allows three-dimensional
imaging with high axial resolution better than 3.8 μm and 5.3 μm in tissue at 800 nm and 1250 nm,
respectively, for precise retinal thickness measurements. A fan-shaped scanning pattern with the pivot point close to the
eye's pupil and a contact lens are applied to obtain optical access to the eye's fundus. First in vivo experiments in a RCS
(royal college of surgeons) rat model with gene-related degeneration of the photoreceptor cells show good visibility of
the retinal microstructure with sufficient contrast for thickness measurement of individual retinal layers. An enhanced
penetration depth at 1250 nm is clearly identifiable revealing sub-choroidal structures that are not visible at 800 nm.
Furthermore, additional simultaneous imaging at 1250 nm improves image quality by frequency compounding speckle
noise reduction. These results are encouraging for time course studies of the rodent retina concerning its development
related to disease progression and treatment response.
The recently introduced new phase-dependent Doppler model for spectral domain optical coherence tomography (SD OCT) has shown that the simple linear relation between the Doppler phase shift and the axial velocity component of an obliquely moving sample is not valid. Additionally, for nearly transverse sample motion with high velocities the phase shift will approach a constant value. Consequently, for small Doppler angles the velocity measurement range of the phase-resolved Doppler analysis is limited in SD OCT. Since these undesirable small Doppler angles can not be prevented, for example, in the in vivo 3-D measurement, we introduce a novel method extending the limited velocity detection range taking the signal power decrease due to fringe washout in SD OCT into account. The signal damping of an obliquely moving sample is presented as a function of the axial and transverse displacement by a universally valid contour plot and does not correspond simply to the sum of the axial and transverse effect. A quantitative combination of the Doppler analysis and the signal-damping method is presented with a flow phantom model. The practicability of this new combined method is presented for the blood flow of the saphenous artery in the in vivo mouse model.
We have recently shown that for any oblique sample movement containing a transverse velocity component, the
prevalent classic Doppler model assuming that the phase shift is proportional to the axial velocity component is
erroneous for spectrometer-based FD OCT. While the previous derivation assumed a continuous integration of the
photocurrent, we extend the new Doppler model for detectors with a shutter control by taking the detector dead time into
account. Because an analytical solution for the new relation between phase shift and oblique sample displacement can
not be given, numerically calculated universal contour plots, which are valid for any center wavelength and beam size,
are presented for detector dead times ranging from 5 % to 90 %. Compared to systems with a duty cycle of 100 %, the
average phase shift does not approach a constant value for large transverse displacements and high sample velocities. In
contrast, at large detector dead times and with this small integration times, the numerically simulated phase shift
corresponds almost to the assumed one according to the classic Doppler model for the investigated velocity range. The
theoretical results were verified by using a flow phantom model.
Reason for using optical coherence tomography (OCT) to locate the germinal disc is the questionable and ethically
alarming killing of male layer chickens because for the layer line only the females are necessary. To avoid this and to
protect the animal rights, the sex of the fertilized chicken egg has to be determined as early as possible in the
unincubated state. Because the information whether the chick becomes male or female can be found in the germinal disc
an accurate localization for sexing is essential. The germinal disc is located somewhere on top of the yolk and has a
diameter of approximately 4 - 5 mm. Different imaging methods like ultrasonography, 3D-X-ray micro computed
tomography and magnetic resonance imaging were used for localization until now, but found to be impractical. The goal
of this study is to prove if OCT can be a moderate approach for the precise in ovo localization. Because the eggshell is an
impenetrable barrier for OCT and to minimize the penetration of germs a very small hole is placed in the eggshell and a
fan-shaped optical scanning pattern is used.
The inhomogeneous backscattering distribution of low-coherent light in blood vessels, which appears as waisted double
fan-shaped intensity pattern, is investigated in an in vivo mouse model and flow phantom measurements using high
resolution spectral domain optical coherence tomography in the 1.3 μm wavelength region. Based on a predicted
orientation of the red blood cells towards laminar flow, an angular modulation of the corresponding backscattering crosssection
inside the vessels is assumed. In combination with the signal attenuation in depth by absorption and scattering, a
simple model of the intravascular intensity modulation is derived. The suitability of the model is demonstrated
exemplarily at the saphenous artery of the mouse during different states of the heart cycle as well as at phantom
measurements with well known flow characteristics. The obtained data and the predicted model show good
correspondence to each other which leads to the conclusion that the red blood cell orientation seems to be the reason for
the observed intensity distribution inside the blood vessels. Therefore, the analysis of the intravascular intensity pattern
might be useful for the evaluation of flow characteristics. Additional investigations of the precise angular backscattering
of the complex shaped red blood cells are necessary for further model refinement.
Resonant Doppler flow imaging based on optical coherence tomography (OCT) is a recently developed imaging
modality that provides, besides the structural information, dynamic blood flow information. We show that this method
can be applied to a common path OCT system by mounting the mirror in the reference arm on a small piezo actor leading
to a simpler and more stable system design. Besides the known 3 state cycle, we describe other cycles with any number
of states leading to higher measurement speed or larger velocity range. The hysteresis of the piezo actor is compensated
by applying an optimized electrical signal. Two different approaches, one using a Levenberg-Marquardt optimization,
the other using the Prandtl-Ishlinskii model for compensation of hysteresis, are applied to generate the optimized control
signal. Besides providing an analytical formula for the calculation of the axial velocity for cycles having certain spacings
in the reference velocity, we describe deviations from the signal degradation caused by the transversal part of the motion
causing errors in the velocity estimation. The performance of the system with two and three states is first evaluated with
a mirror on a loud speaker. Measurements with a flow phantom consisting of 1 % Intralipid dilution flowing through
small diameter capillaries show the suitability of the system and the expected deviations at high velocities.
KEYWORDS: Lung, Optical coherence tomography, 3D image processing, In vivo imaging, Tissues, Injuries, Imaging systems, 3D acquisition, Doppler tomography, Scanners
In this feasibility study we present a method for 4D imaging of healthy and injured subpleural lung tissue in a mouse
model. We used triggered swept source optical coherence tomography with an A-scan frequency of 20 kHz to image
murine subpleural alveoli during the ventilation cycle. The data acquisition was gated to the pulmonary airway pressure
to take one B-scan in each ventilation cycle for different pressure levels. The acquired B-scans were combined offline to
one C-scan for each pressure level. Due to the high acquisition rate of the used optical coherence tomography system, we
are also able to perform OCT Doppler imaging of the alveolar arterioles. We demonstrated that OCT is a useful tool to
investigate the alveolar dynamics in spatial dimensions and to analyze the alveolar blood flow by using Doppler OCT.
Optical coherence tomography (OCT) is performed in the spectral domain simultaneously at two different wavelength
bands centered at 800 nm and 1250 nm. A novel commercial supercontinuum laser is applied as a single light source
whose emission spectrum is shaped by optical and spatial filtering to obtain an adequate double peak spectrum. After
spectral shaping, the wavelength bands 700 - 900 nm and 1100 - 1400 nm are used for OCT imaging. A fiber-coupled
setup optimized for both spectral regions facilitates easy and flexible access to the measurement area. Each wavelength
band is analyzed with an individual spectrometer at an A-scan rate of about 12 kHz which allows real-time sample
examination. The free-space axial resolutions were measured to be less than 4.5 μm and 7 μm at 800 nm and 1250 nm,
respectively. This technique combines the high resolution at 800 nm with the enhanced imaging depth at 1250 nm.
Furthermore, spatially resolved spectroscopic sample features are extracted by comparing the backscattering properties at
the two different wavelength bands, showing the ability of dual-band OCT to enhance image contrast.
Blood flow measurement with spectrometer-based Fourier domain optical coherence tomography (FD OCT) is limited
by the motion-induced signal fading and the resulting reduction of flow sensitivity. In this study, a combination of the
established Doppler OCT and the numerically simulated signal damping due to obliquely moved scatterers is used to
estimate the systolic blood flow velocities in the in vivo mouse model at which the standard Doppler OCT does not work
any longer.
We present a novel method to obtain optical angiographies (OAG) on a standard optical coherence tomography
(OCT) system. The moving reference arm is simulated by introducing a phase-shift at the post-processing stage.
The method can be applied bi-directionally from a single scan, one or more velocity-thresholds can be adjusted
during post-processing. First in-vivo results are shown.
In this study, we demonstrate that phase-resolved Doppler optical frequency domain imaging (OFDI) is very suitable to
quantify the pulsatile blood flow within a vasodynamic measurement in the in vivo mouse model. For this, an OFDI-system
with a read-out rate of 20 kHz and a center wavelength of 1320 nm has been used to image the time-resolved
murine blood flow in 300 μμm vessels. Because OFDI is less sensitive to fringe washout due to axial sample motion, it is
applied to analyze the blood flow velocities and the vascular dynamics in six-week-old C57BL/6 mice compared to one
of the LDLR knockout strain kept under sedentary conditions or with access to voluntary wheel running. We have shown
that the systolic as well as the diastolic phase of the pulsatile arterial blood flow can be well identified at each
vasodynamic state. Furthermore, the changes of the flow velocities after vasoconstriction and -dilation were presented
and interpreted in the entire physiological context. With this, the combined measurement of time-resolved blood flow
and vessel diameter provides the basis to analyze the vascular function and its influence on the blood flow of small
arteries of different mouse strains in response to different life styles.
In-vivo imaging of the vascular system can provide novel insight into the dynamics of vasoconstriction and vasodilation. Fourier domain optical coherence tomography (FD-OCT) is an optical, noncontact imaging technique based on interferometry of short-coherent near-infrared light with axial resolution of less than 10 µm. In this study, we apply FD-OCT as an in-vivo imaging technique to investigate blood vessels in their anatomical context using temporally resolved image stacks. Our chosen model system is the murine saphenous artery and vein, due to their small inner vessel diameters, sensitive response to vasoactive stimuli, and advantageous anatomical position. The vascular function of male wild-type mice (C57BL/6) is determined at the ages of 6 and 20 weeks. Vasoconstriction is analyzed in response to dermal application of potassium (K+), and vasodilation in response to sodium nitroprusside (SNP). Vasodynamics are quantified from time series (75 sec, 4 frames per sec, 330×512 pixels per frame) of cross sectional images that are analyzed by semiautomated image processing software. The morphology of the saphenous artery and vein is determined by 3-D image stacks of 512×512×512 pixels. Using the FD-OCT technique, we are able to demonstrate age-dependent differences in vascular function and vasodynamics.
Spectrometer-based or space encoded Fourier domain OCT is preferred because of its relatively simple design, fast
measurement speed and good signal-to-noise ratio. Besides delivering structural information, it is often used to measure
blood flow velocities. Commonly, the axial component of the velocity is calculated from the phase difference of
consecutive A-Scans. While this result holds true for pure axial movement, a transversal component of the displacement
will alter this simple relationship. We present a new model accounting for the changing intensity of the illuminating
beam on the moving particles and explaining why the phase difference does not increase linearly with the velocity.
Movements as small as 20 % of the beam diameter during the integration time of the line detector will alter the observed
phase shifts noticeably. For small angles between transversal direction and direction of movement, the discrepancy
between classically calculated and measured phase shift may be huge. At certain velocities and angles no correlation of
the phase exists even so there is an OCT-signal. High velocities at small angles will result in a limit for the phase shift
smaller than π. A safe region, where the deviations to the linear relationship between axial velocity and phase shift are
small, is specified.
Blood flow measurement with spectrometer-based Fourier domain optical coherence tomography (FD-OCT) is limited
by the motion-induced signal fading and the resulting reduction of flow sensitivity. Therefore, we have numerically
simulated the signal power decrease of an obliquely moved scattering layer as a function of the absolute sample velocity
composed of an axial and transverse component. In contrast to the prevalent expectance, the resulting signal damping is
not only the sum of axial and transverse effect. In this study, we take advantage of the signal decay and present the feasibility to quantify high flow velocities at which the standard Doppler OCT does not work any longer. For the validation of our approach, a flow phantom model consisting of a 1%-Intralipid solution and a 320 μm glass capillary was used. With this phantom study, depth-resolved flow was visualized and the quantitative velocities were extracted from the OCT images without phase information.
KEYWORDS: Optical coherence tomography, Blood vessels, Resistance, Potassium, In vivo imaging, Scanners, Head, 3D image processing, Image analysis, Near infrared
In vivo imaging of blood vessels obtain useful insights in characterizing the dynamics of vasoconstriction and
vasodilation. Fourier domain optical Coherence Tomography (FD-OCT) imaging technique permits in vivo investigation
of blood vessels in their anatomical context without preparation traumata by temporal resolved image stacks. OCT is an
optical, contact less imaging technique based on Michelson interferometry of short coherent near infrared light.
Particularly by the possibility of a contact-less measurement and the high axial resolution up to 10 microns OCT is
superior to an investigation by ultra sound measurement. Furthermore we obtain a high time resolution of vessel dynamic
measurements with the used Fourier domain OCT-system by a high A-scan rate [1,22kHz]. In this study the model of
saphenous artery was chosen for analyzing function and dynamics. The arteria saphena in the mouse is a suitable blood
vessel due to the small inner diameter, a sensitive response to vasoactive stimuli and an advantageous anatomically
position. Male wild type mice (C57BL/6) at the age of 8 weeks were fed control or high-fat diet for 10 weeks before
analyzing the vasodynamics. The blood vessel was stimulated by dermal application of potassium to induce
vasoconstriction or Sodium-Nitroprusside (SNP) to induce vasodilation. The morphology of the a. saphena and vein was
determined by 3D image stacks. Time series (72 seconds, 300x512 pixel per frame) of cross-sectional images were
analysed using semi automatic image processing software. Time course of dynamic parameters of the vessel was
measured.
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