Calibrating intensity values of optical coherence tomography (OCT) images helps to extract valuable tissue information, such as the attenuation coefficient. To achieve this, knowledge of focus position and Rayleigh length is crucial. We study the potential of obtaining the focus position from an OCT scan acquired with a single focus setting using the system’s longitudinal chromatic aberration. Different focus positions for sub-spectrum reconstructed OCT images are realized by taking advantage of the chromatic focal shift. Ratios of these sub-spectral images are used to estimate the focus position. High-resolution B-scan reconstruction is demonstrated by coherently combining sub-spectrum confocal function corrected B-scans, followed by OCT attenuation coefficient imaging. Additionally, we present an approach to experimentally identify the chromatic focal shift from OCT data itself.
Quantitative OCT measurements of optical parameters such as the attenuation coefficient could allow for better differentiation between healthy and diseased tissues. To investigate relationship between the attenuation coefficient and OCT signal we developed a Monte Carlo (MC) simulation method for OCT measurements that takes into account the confocal function. To validate the MC model simulations were compared to OCT measurements on phantoms. Both the A-lines, and attenuation coefficients extracted from the A-lines based on a singly scattered model, matched well between the simulated and experimentally measured data.
KEYWORDS: Monte Carlo methods, Photons, Scattering, Optical coherence tomography, Signal attenuation, Tissues, Optical phantoms, Tissue optics, Signal detection
Optical coherence tomography (OCT) is a technique that is widely used to diagnose pathologies through morphological changes in tissue structure. OCT imaging of such structures based on the backscattered intensity provides qualitative images that are not quantitatively related to the tissue’s optical properties. Since certain pathologies affect optical parameters of tissues, such as the attenuation coefficient, quantification of the attenuation coefficient could be useful for medical diagnosis. For accurate quantisation of optical parameters robust depth resolved OCT models which are valid over a wide range of parameters are necessary.
The attenuation coefficient can be calculated from OCT data, but accurate determination requires compensating for the confocal function. We present detailed measurement series for extraction of the focal plane and the apparent Rayleigh length from the ratios of OCT images acquired with different focus depths and compare these results with alternative approaches. The optimal focus depth difference is determined for intralipid and titanium oxide phantoms with different scatterer concentrations and the attenuation coefficients corrected for the confocal function are calculated. We further demonstrate good reproducibility of the determined attenuation coefficient of layers with identical scatter concentrations in a multi-layered titanium oxide phantom.
KEYWORDS: Reflectivity, Luminescence, Confocal microscopy, In vivo imaging, Retinal scanning, Fluorescence lifetime imaging, Retina, Signal detection, Visualization, Imaging systems
We have developed a compact hollow core fiber (HCF)-based imaging platform capable of simultaneous in vivo confocal reflectance and two-photon imaging through the mouse pupil. We demonstrate the performance of this platform by imaging retinal ganglion cells (RGCs) in which the fluorophores YFP and GCaMP3 are expressed in Thy1-YFP-16 and Thy1-GCaMP3 transgenic mice, respectively. Confocal reflectance images of the mouse retina served as a reference for the simultaneous acquisition of the two-photon signals that clearly showed RGCs with single-cell resolution. The use of an HCF platform makes the system compact with future application in the longitudinal investigation into the structure and function of healthy and diseased RGCs.
Time-resolved video photographs have been taken in order to investigate plasma induced surface ablation of soft tissue by IR picosecond laser pulses with energies of about 1 mJ. The emission and propagation of shock waves in the irradiated tissue as well as in the surrounding air environment was studied. The pressure amplitudes of the shock transients were determined from the measured shock velocities. A decay of the pressure amplitude below 100 MPa was observed within a distance of about 200 micrometers from the center of laser induced optical breakdown. The dynamics of the ablation crater and the ejection of the ablation fragments was studied on a larger time scale. The maximum expansion of the ablation crater was measured to be about 200 (Mu) m at temporal delays of 4-5 microsecond(s) after the impact of the laser pulse. Furthermore, generation and propagation of a surface deformation wave was observed. Thus, we present a detailed and consistent description of all phenomena occurring during plasma-mediated surface ablation of soft tissue.
Three nonlinear (beta) -barium borate (BBO) crystals were used to generate the fourth and fifth harmonic of a Nd:YLF picosecond laser with a fundamental wavelength of 1053 nm. The UV laser pulses were used to study the surface ablation of bovine soft tissue. Whereas the general study was concentrated on brain tissue, first results were also obtained with cornea tissue. The ablation efficiency as a function of pulse energy and the ablation threshold were determined for both UV wavelengths. Histology and SEM- microscopy of the performed laser lesions are presented. Furthermore, time-resolved flash photographs we recorded by illuminating the ablation area with green picosecond laser pulses and detecting the probe beam with a CCD chip. The photographs yielded information about the physical properties of the ablation process. The emission of a shock wave into the air environment as well as the ejection of dissected ablation fragments was observed for pulse energies down to 10 (mu) J. Both UV wavelengths can be combined with an ophthalmic application system, which comprises an xyz- scanning unit for realizing precalculated ablation patterns and a z-tracker for detecting the current surface position of the cornea. The information of the z-tracker can be used to compensate the axial eye-movement with the xyz-scanning unit.
The phase retardation of light induced by the birefringent parts of the human retina in vivo is measured with an electro-optical ellipsometer using the principle of confocal imaging. A scanning unit allows to examine an area of 25 degrees by 12.5 degrees on the retina with a resolution of 256 by 128 points with both, incident and exit beam transmitting the cornea at a fixed position. Due to this fixed beam position the Muellermatrix of the cornea can be calculated using light which is specularly reflected on blood vessels lying above the nerve fiber layer of the retina. The obtained images show a homogenous radial distribution of the retinal retardation around the fovea without the appearance of the so called Haidinger brushes. The areas with the thickest nerve fiber layers, the arcuate bundles, appear in their typical arc and were measured quantitatively. In addition, an alternative method for the compensation of corneal birefringence is evaluated by focusing the light beam onto the surface of the lens. Hereby, the measured area in the center of the cornea is 3 X 0.75 mm2.
In-vitro ablation experiments of calf brain tissue with ultrashort lasers are reported. Excisions have been performed by the mechanism of plasma-mediated ablation with focused picosecond laser pulses. Results achieved with a 35 ps oscillator/regenerative amplifier Nd:YLF laser at 1053 nm and its second and fourth harmonic at 526 and 263 nm, respectively, are presented. Preliminary experiments with 19 ps laser pulses at wavelengths near 3 micrometers generated by optical parametric amplification are reported. Here, during the ablation with low photon energies no plasma was induced. Furthermore, the spectrum of laser-induced plasma sparks on the surface of fresh calf brain tissue was recorded. Atomic line widths were measured and used to determine the electron density of the plasma. With energy densities of about 100 J/cm2 values between 2 (DOT) 1017 and 5 (DOT) 1018 cm-3 were found.
Plasma mediated ablations of calf brain tissue and corneal tissue have been performed with ultrashort pulses provided by different laser systems: A 30 ps Nd:YLF amplified laser with up to 1.5 mJ of pulse energy and a powerful 180 fs Ti:Sapphire oscillator/regenerative amplifier laser system capable of pulse energies of up to 300 (mu) J. With the femtosecond pulses the threshold in energy density needed for initiating the ablation process was found to be significantly lower than with longer pulse durations. Furthermore, the amount of ablated material is by a factor of two greater with the femtosecond pulses compared to the results obtained with the picosecond pulses. Histological examinations did not show any thermal or structural damage in adjacent tissue with pulses in the pico- and femtosecond range as it could be found after irradiation with longer pulses. The morphology of the excisions is of a very high quality.
A new method for the fast measurement of the Z-transferfunction in the Fovea Centralis of the human eye using a confocal laser scanning device is presented. The tested eye is illuminated by a collimated helium-neon laser beam and therefore focuses the light onto the retina. Reflected light is detected using confocal techniques, i.e. a pinhole in the detection unit assures that only light originating from the focal plane is sampled. The focus of the laser beam is scanned in 1-D along the visual axis of the eye through the regina by adding a slide defocus to the incoming beam. The Z-transferfunction can be obtained directly by measuring the intensity of the reflected light for different focal planes. A fixation target is offered to the eye at infinity to stabilize the accommodation as well as the direct viewing. Different pinholes in the detection unit are tested to investigate signal to noise ratio and depth resolution.
In vitro ablation experiments of calf brain tissue using a picosecond Nd:YLF laser system are reported. The laser lesions were studied with special regard to the ablation rate and to possible injuries of tissue adjacent to the excisions. Histological examinations prove the excellent quality of the lesions, showing no signs of thermal damages or carbonization. Furthermore, the influence of higher photon energies on the ablation threshold was studied.
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