Maximiliano Cuevas

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.

Regional lung aeration and ventilation during pressure support and biphasic positive airway pressure ventilation in experimental lung injury

IntroductionThere is an increasing interest in biphasic positive airway pressure with spontaneous breathing (BIPAP+SBmean), which is a combination of time-cycled controlled breaths at two levels of continuous positive airway pressure (BIPAP+SBcontrolled) and non-assisted spontaneous breathing (BIPAP+SBspont), in the early phase of acute lung injury (ALI). However, pressure support ventilation (PSV) remains the most commonly used mode of assisted ventilation. To date, the effects of BIPAP+SBmean and PSV on regional lung aeration and ventilation during ALI are only poorly defined.MethodsIn 10 anesthetized juvenile pigs, ALI was induced by surfactant depletion. BIPAP+SBmean and PSV were performed in a random sequence (1 h each) at comparable mean airway pressures and minute volumes. Gas exchange, hemodynamics, and inspiratory effort were determined and dynamic computed tomography scans obtained. Aeration and ventilation were calculated in four zones along the ventral-dorsal axis at lung apex, hilum and base.ResultsCompared to PSV, BIPAP+SBmean resulted in: 1) lower mean tidal volume, comparable oxygenation and hemodynamics, and increased PaCO2 and inspiratory effort; 2) less nonaerated areas at end-expiration; 3) decreased tidal hyperaeration and re-aeration; 4) similar distributions of ventilation. During BIPAP+SBmean: i) BIPAP+SBspont had lower tidal volumes and higher rates than BIPAP+SBcontrolled; ii) BIPAP+SBspont and BIPAP+SBcontrolled had similar distributions of ventilation and aeration; iii) BIPAP+SBcontrolled resulted in increased tidal re-aeration and hyperareation, compared to PSV. BIPAP+SBspont showed an opposite pattern.ConclusionsIn this model of ALI, the reduction of tidal re-aeration and hyperaeration during BIPAP+SBmean compared to PSV is not due to decreased nonaerated areas at end-expiration or different distribution of ventilation, but to lower tidal volumes during BIPAP+SBspont. The ratio between spontaneous to controlled breaths seems to play a pivotal role in reducing tidal re-aeration and hyperaeration during BIPAP+SBmean.

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.

Characteristics of highly flexible PDMS membranes for long-term mechanostimulation of biological tissue.

Measurement of mechanical properties of soft biological tissue remains a challenging task in mechanobiology. Recently, we presented a bioreactor for simultaneous mechanostimulation and analysis of the mechanical properties of soft biological tissue samples. In this bioreactor, the sample is stretched via deflection of a flexible membrane. It was found that the use of highly compliant membranes increases accuracy of measurements. Here, we describe the production process and characteristics of thin and flexible membranes of polydimethylsiloxane (PDMS) designed to improve the signal-to-noise ratio of our bioreactor. By a spin-coating process, PDMS membranes were built by polymerization of a two component elastomer. The influence of resin components proportion, rotation duration, and speed of the spinning were related to the membrane mechanics. Membranes of 22 mm inner diameter and 33 to 36 microm thickness at homogeneous profiles were produced. Isolated rat diaphragms served as biological tissue samples. Mechanical properties of the membranes remained constant during 24 h of mechanostimulation. In contrast, time- and strain-dependent mechanical properties of the diaphragms were found.

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.

Resonant Doppler imaging with common path OCT

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.

Analysis of murine vascular function in vivo by optical coherence tomography in response to high-fat diet.

In this study, we demonstrate the application of optical coherence tomography (OCT) as a contactless imaging technique to analyze vasodynamics in small blood vessels in vivo. The transluminal OCT imaging of vessels avoids micro traumata in the endothelium and circumvents surgical intervention. It can be performed in the intact perfused vessel and provides a new method to measure vascular function and dynamics in vivo. The resolution of 10 mum and the velocity of image acquisition are adequate to detect differences in the inner diameter, the maximal velocity, or the time to half-maximal diameter change of small vessels. We applied this new technology to study the vascular dynamics in small vessels of 6- and 20-week-old C57BL/6 mice in vivo. In addition, we determined by this technique the impact of a high-fat diet for 14 weeks on vascular function in 20-week-old animals. The diameter of the saphenous artery was increased under resting conditions, after vasoconstriction and after vasodilatation in 20-week-old animals on normal chow and high-fat diet, compared to 6-week-old animals. High-fat diet caused a significantly impaired vasoconstriction in the saphenous artery. The maximal velocity of diameter changes of the saphenous artery was determined by time-resolved OCT imaging. A significant reduction of this parameter was found during vasoconstriction in 20-week-old mice on high-fat diet, compared to 6-week-old animals. In conclusion, transluminal optical coherence tomography imaging is a novel and useful technique to analyze the impaired vasodynamics of small arteries in response to high-fat diet in vivo.

Simultaneous dual-band spectral domain optical coherence tomography using a supercontinuum laser light source

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.

Measurement of the Epidermal Thickness with Fourier Domain Optical Coherence Tomography in an Occupational Health Study on Natural Sun Protection of the Human Skin

Fourier domain optical coherence tomography (FD-OCT) was used to take cross-sectional images of the skin at the forehead, back of hand and back of volunteers. The aim of the occupational health study was the investigation of the natural sun protection of outdoor workers and in particular the quantification of the UV induced epidermal thickening. A set of manually segmented OCT images was used as a gold standard to evaluate the performance of automatic algorithms for finding the epidermal thickness. The established A-scan averaging method and two newly developed automatic algorithms were compared. The first of the new methods is based on Markov random chains and the second new and best performing algorithm is based on a convolution filter with a trace finding routine with a quadratic cost function. Although the results of all algorithms differ in the absolute values for the epidermal thickness, they all show the same tendencies. There is a strong dependency of epidermal thickness on skin site, a weak dependency on the season of the year and a noticeable epidermal thickening 7 days of recovery after UV induced erythema.

Fabrication of thin and flexible PDMS membranes for biomechanical test applications

For continuous analysis of mechanical properties of soft biological tissues during mechanical stimulation we developed a bioreactor to apply controlled periodic deflection to a probe. The probe attached on an elastic membrane produces a counterpressure if a certain gas volume is applied to the pressure chamber under the membrane. The related counterpressure-volume relationship describes the mechanical properties of the membrane-probe combination. To improve the signal-to-noise ratio of measurements the use of highly flexible membranes is advantageous.