Julia Walther

Simultaneous dual-band optical coherence tomography in the spectral domain for high resolution in vivo imaging

Optical coherence tomography (OCT) in the spectral domain is demonstrated simultaneously at two wavelength bands centered at 800 nm and 1250 nm. A novel commercial supercontinuum laser is applied as a single low coherence broadband light source. The emission spectrum of the source is shaped by optical and spatial filtering in order to achieve an adequate double peak spectrum containing the wavelength bands 700 - 900 nm and 1100 - 1400 nm for dual-band OCT imaging and thus reducing the radiation exposure of the sample. Each wavelength band is analyzed with an individual spectrometer at an A-scan rate of about 12 kHz which enables real-time imaging for the examination of moving samples. A common path optical setup optimized for both spectral regions with a separate single fiber-based scanning unit was realized which facilitates flexible handling and easy access to the measurement area. The free-space axial resolutions were measured to be less than 4.5 microm and 7 microm at 800 nm and 1250 nm, respectively. Three-dimensional imaging ten times faster than previously reported with a signal-to-noise-ratio of above 90 dB is achieved simultaneously in both wavelength bands. Spectral domain dual-band OCT combines real-time imaging with high resolution at 800 nm and enhanced penetration depth at 1250 nm and therefore provides a well suited tool for in vivo vasodynamic measurements. Further, spatially resolved spectral features of the sample are obtained by means of comparing the backscattering properties at two different wavelength bands. The ability of dual-band OCT to enhance tissue contrast and the sensitivity of this imaging modality to wavelength-dependent sample birefringence is demonstrated.

Optical coherence tomography in biomedical research.

Optical coherence tomography (OCT) is a noninvasive, high-resolution, interferometric imaging modality using near-infrared light to acquire cross-sections and three-dimensional images of the subsurface microstructure of biological specimens. Because of rapid improvement of the acquisition speed and axial resolution of OCT over recent years, OCT is becoming increasingly attractive for applications in biomedical research. Therefore, OCT is no longer used solely for structural investigations of biological samples but also for functional examination, making it potentially useful in bioanalytical science. The combination of in vivo structural and functional findings makes it possible to obtain thorough knowledge on basic physiological and pathological processes. Advanced applications, for example, optical biopsy in visceral cavities, have been enabled by combining OCT with established imaging modalities. This report gives an outline of the state of the art and novel trends of innovative OCT approaches in biomedical research in which the main focus is on applications in fundamental research and pre-clinical utilization.

Shear flow-induced optical inhomogeneity of blood assessed in vivo and in vitro by spectral domain optical coherence tomography in the 1.3 μm wavelength range

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.

Transverse motion as a source of noise and reduced correlation of the Doppler phase shift in spectral domain OCT

Recently, a new phase-resolved Doppler model was presented for spectral domain optical coherence tomography (SD OCT) showing that the linear relation between the axial velocity component of the obliquely moving sample and the phase difference of consecutive A-Scans does not hold true in the presence of a transverse velocity component which is neglected in the widely-used classic Doppler analysis. Besides taking note of the new non-proportional relationship of phase shift and oblique sample motion, it is essential to consider the correlation of the phase shift and its specific characteristic at certain Doppler angles for designing Doppler experiments with SD OCT. A correlation quotient is introduced to quantify the correlation of the backscattering signal in consecutive A-Scans as a function of the oblique sample motion. It was found that at certain velocities and Doppler angles no correlation of the phases of sequential A-Scans exists, even though the signal does not vanish. To indicate how the noise of the Doppler phase shift behaves for oblique movement, the standard deviation is determined as a function of the correlation quotient and the number of complex Doppler data averaged. The detailed theoretical model is validated by using a flow phantom model consisting of a 1% Intralipid flow through a 310 microm capillary. Finally, a short discussion of the presented results and the consequence for performing Doppler experiments is given.

Effects of axial, transverse, and oblique sample motion in FD OCT in systems with global or rolling shutter line detector

This study deals with effects on the interference signal caused by axial, transverse, and oblique motion in spectrometer-based Fourier-domain optical coherence tomography (FD OCT). Two different systems are compared-one with a global shutter line detector and the other with a rolling shutter. We present theoretical and experimental investigations of motion artifacts. Regarding axial motion, fringe washout is observed in both systems, and an additional Doppler frequency shift is seen in the system using a rolling shutter. In addition, both systems show the same SNR decrease as a result of a transversely and obliquely moving sample. The possibility of flow measurement by using the decrease in signal power was demonstrated by imaging 1% Intralipid emulsion flowing through a glass capillary. This research provides an understanding of the SNR degradation caused by sample motion and demonstrates the importance of fast data acquisition in medical imaging.

Investigation of murine Vasodynamics by Fourier Domain Optical Coherence Tomography

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.

Endoscopic optical coherence tomography device for forward imaging with broad field of view

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.

Signal power decrease due to fringe washout as an extension of the limited Doppler flow measurement range in spectral domain optical coherence tomography

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.

Relation of joint spectral and time domain optical coherence tomography (jSTdOCT) and phase-resolved Doppler OCT

A variety of promising approaches for quantitative flow velocity measurement in OCT have been proposed in recent years. The question is: Which method gets the most precise flow velocity out of the interference signals detected. We have compared the promising joint spectral and time domain optical coherence tomography (jSTdOCT) and the commonly used phase-resolved Doppler OCT (DOCT) and describe the link between these two proven methods for OCT in the Fourier domain (FD OCT). First, we show that jSTdOCT can be significantly improved by calculating the center of gravity via an unbiased complex algorithm instead of detecting the maximum intensity signal of the broadened Doppler frequency spectrum. Secondly, we introduce a unified mathematical description for DOCT and jSTdOCT that differs only in one exponent and call it enhjSTdOCT. Third, we present that enhjSTdOCT has the potential to significantly reduce the noise of the velocity measurement by choosing an exponent depending on the transverse sample velocity component and the signal-to-noise ratio. EnhjSTdOCT is verified numerically and experimentally to find the optimal parameters for maximal velocity noise reduction.

In-vivo Fourier domain optical coherence tomography as a new tool for investigation of vasodynamics in the mouse model

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 microm. 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 x 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 x 512 x 512 pixels. Using the FD-OCT technique, we are able to demonstrate age-dependent differences in vascular function and vasodynamics.