project, which is funded by the German Federal Ministry of Education and Research, attempts to improve the accessibility of diagnostic instruments for glaucoma screening. The presented approach aims to realize real-time nearinfrared video fundus imaging that enables the use of targeted fixation stimuli to ensure continuous imaging. The integration of near-infrared illumination with a wavelength of 780 nm not only avoids pupil constriction, but also enables mesopic imaging in darkened ambient light, ensuring optimal visualization of the retinal structure. This innovative system achieves nearly reflection-free imaging through polarized illumination with polarization-dependent beam paths. Its primary aim is to capture extensive fundus areas to facilitate correlations with linear optical coherence tomography (LOCT) measurements. In the future, the fundus setup will be integrated into the LOCT setup. In this research project, the primary aim is to generate images of the optic nerve, but it is also possible to carry out examinations of the macula. Unlike from traditional fundus cameras, this system has a controllable screen for generating individual fixation stimuli, which creates continuous eye movements and enables controlled imaging. The main objective is to capture large fundus areas and track eye positions to combine this information with the LOCT measurements A-scan positions, which enables the creation of B-scans with irregular geometries. This approach replaces the need for complex scanning systems by leveraging natural eye movements. The approach can thus be used to detect retinal pathologies in a different way and could therefore be used for more comprehensive diagnostic and scientific applications.
The accommodation of the crystalline lens allows for sharp vision in the close and distant range. Conditions such as cataracts can make it necessary to replace the natural tissue with an artificial intraocular lens (IOL), which lacks the ability to accommodate. The alternatives that are currently under investigation include accommodating IOLs or refilling the lens bag with hydrogels. Here, we investigate the possibility to remove and refill only the central part (nucleus) of a cataract lens, thereby preserving its ability to accommodate. This approach avoids damage to the lens cortex to prevent stiffening of the capsular bag—a significant drawback of complete lens refilling. The nucleus of the lenses of porcine eyes was fragmented via fs-laser treatment and removed by phacoemulsification. The lens’s mechanical properties, ray tracing properties and curvature were investigated with an in-house developed measurement setup, including a lens stretching device for simulation of accommodation. This yielded quantifiable data on the transparency, accommodation capabilities and focus shift of treated versus untreated lenses. While native transparency could not yet be achieved, refilled eyes exhibited the same focal shift as non-refilled, indicating functional accommodation. Measurements of the curvature revealed stronger flattening of refilled eyes. Apart from the study of partial lens refill, the stretcher device and respective protocols presented here possess great potential in IOL development, presbyopia research or characterization of model lenses.
Femtosecond laser oscillator systems with low pulse energy (< 1 μJ) and high repetition rate (MHz) are increasingly used for precise, fast and safe eye surgery. Therefore, the laser tissue interaction process is of great interest to optimize and improve established and future surgical protocols. Besides, using faster laser systems leads to unintended self-induced interaction effects, where a femtosecond laser pulse modifies the vicinity in the material in such a way that the focus of following laser pulses is changed. We used a femtosecond oscillator laser system with high repetition rate and 66 nJ pulse energy to produce photodisruption in water. Water was used as phantom material for ocular tissue, because tissue mainly consists of water. A custom made digital-holographic system was used to measure the temporal material modification from picoseconds until seconds after occurrence of the photodisruption. For illumination of the sample we used either a continuously light source or the femtosecond laser pulse itself in a pump-probe configuration. The holographic system provides quantitative data of phase difference Δφ for the full field of view of several tenth of micrometers. Phase difference is equivalent to the laser induced change in the material’s refractive index which can alter focusing conditions of following laser pulses and might impair surgical outcome. We obtained the largest change in Δφ during the first picoseconds, followed by a slow relaxation of Δφ within some milliseconds. The results of time resolved measurements of the laser induced material modification will help to optimize scanning schemes in ocular surgery.
We present a time-resolved photographic analysis of the pulse-to-pulse interaction. In particular, we studied the influence of the cavitation bubble induced by a fs-pulse on the optical focusing of the consecutive pulse and its cavitation bubble dynamics in dependence on temporal pulse separation in water. As a first result, by decreasing the temporal separation of laser pulses, there is a diminishment of the laser-induced optical breakdown (LIOB) efficiency in terms of energy conversion, caused by disturbed focusing into persisting gas bubbles at the focal volume. A LIOB at the focal spot is finally suppressed by impinging the expanding or collapsing cavitation bubble of the preceding pulse. These results could be additionally confirmed in porcine gelatin solution with various concentrations. Hence, the interaction between the laser and transparent ophthalmic tissue may be accompanied by a raised central laser energy transmission, which could be observed in case of a temporal pulse overlap. In conclusion, our experimental results are of particular importance for the optimization of the prospective ophthalmic surgical process with future generation fs-lasers.
The immediate pulse-to-pulse interaction becomes more and more important for future-generation high-repetition rate ophthalmic laser systems. Therefore, we investigated the interaction of two laser pulses with different spatial and temporal separation by time-resolved photography. There are various different characteristic interaction mechanisms which are divided into 11 interaction scenarios. Furthermore, the parameter range has been constricted regarding the medical application; here, the efficiency was optimized to a maximum jet velocity along the scanning axis with minimum applied pulse energy as well as unwanted side effects at the same time. In conclusion, these results are of great interest for the prospective optimization of the ophthalmic surgical process with future-generation fs-lasers.
High repetition rated femtosecond laser oscillator systems with low pulse energy are more often applied for precise and safer eye surgery. Especially, the cutting procedure in the crystalline lens is of high important for presbyopia treatment. Nevertheless, the fundamental laser tissue interaction process is not completely understood, because apparently a self-induced process takes place, were one modified region changes the focusing behavior of following laser pulses. We used a MHz repetition rate femtosecond laser system with nJ-pulse energy which were focused inside an ocular-tissue-phantom (Hydroxy-ethylmethacrylat - HEMA) to induce photodisruption. The material change, caused by the fs-pulses was measured simultaneously with a compact digital-holographic microscope. To investigate the material manipulation at different time scales, we used a continuously illuminating light source. The holographic images provide quantitative values for optical path length difference (OPL), which is equivalent to a refractive index change. This change of the optical properties may cause following pulses to obtain different focusing conditions. Time lapse measurements during the laser application were performed, which show the temporal evolution of OPL. An increase of OPL during the laser application was measured, which was followed by a decrease in OPL after laser processing. Furthermore, similar experiments were performed in distilled water and in native porcine crystalline lenses. The fs-laser cutting effects in HEMA and crystalline lens were transferable. Simultaneous measurements of the material modification during the cutting process give rise to better knowledge of treatment modalities during ocular tissue processing.
We present a time-resolved photographic analysis of the pulse-to-pulse interaction of temporally separated fs-laser pulses with various pulse overlap in water. Initially, by decreasing the temporal separation of laser pulses there is a diminishment of the laser-induced optical breakdown (LIOB) probability, caused by focusing into persisting gas bubbles at the focal volume. A LIOB at the focal spot is finally impaired by the oscillating cavitation bubble of the preceding pulse. Hence, the interaction between the laser and transparent ophthalmic tissue may be accompanied by a raised laser energy transmission and a variation in the axial cutting depth, which could be observed in case of a pulse overlap. In conclusion, our experimental results are of particular importance for the optimization of the prospective ophthalmic surgical process with future-generation fs-lasers.
In ophthalmic surgery fs laser pulses are used as a precise and safe cutting tool. At specific processing parameters, however, an interesting phenomenon of unintended periodic structures inside the tissue can be observed. In this study, a transparent polymer served as ocular phantom material for further investigations. A Femtosecond laser with MHz-repetition rate and pulse energies below 200 nJ was used. The size of the durable material change caused by applied fs-single pulses was measured in regard to the pulse energy. Furthermore, lines were cut inside the material with different laser spot distances and laser pulse energies. The creation and enhancement of unintended step-like structures could be related to a decrease of spot distance and further increase of pulse energy. Cutting planes inside the material resulted also in a formation of step like structures. For planes the step-like structures were formed with different orientation in the x-y-plane in regard to the used line distance between two applied lines. A maximum step height of 75 μm was measured using our setup. Those periodic structures are unwanted for any application in the field of laser material processing, ophthalmology or biomedical sectioning. Hence, investigations of the parameters which trigger this phenomenon are of great interest.
Interaction of subsequent laser pulses becomes important relevant with the use of high-repetition rate fs-laser systems for
ophthalmic laser surgery. Therefore, we investigated the interaction of temporally separated laser pulses in water by
time-resolved photography. With decreasing temporal separation of pulses the probability of laser-induced optical
breakdown (LIOB) is firstly diminished by disturbed focusing into persisting gas bubbles. Finally, LIOB is totally
impaired by the expanding or collapsing cavitation of the preceding pulse. Hence, laser-tissue interaction might be
accompanied by a raised laser energy transmission. In conclusion, these results are of great interest for the prospective
optimization of the ophthalmic surgical process with modern fs-lasers.
A today well-known laser based treatment in ophthalmology is the LASIK procedure which nowadays includes cutting
of the corneal tissue with ultra-short laser pulses. Instead of disposing a microkeratome for cutting a corneal flap, a
focused ultra-short laser pulse is scanned below the surface of biological tissue causing the effect of an optical
breakdown and hence obtaining a dissection. Inside the tissue, the energy of the laser pulses is absorbed by non-linear
processes; as a result a cavitation bubble expands and ruptures the tissue. Hence, positioning of several optical
breakdowns side by side generates an incision. Due to a reduction of the amount of laser energy, with a moderate
duration of treatment at the same time, the current development of ultra-short pulse laser systems points to higher
repetition rates in the range of even Megahertz instead of tens or hundreds of Kilohertz. In turn, this results in a pulse
overlap and therefor a probable occurrence of interaction between different optical breakdowns and respectively
cavitation bubbles of adjacent optical breakdowns. While the interaction of one single laser pulse with biological tissue is
analyzed reasonably well experimentally and theoretically, the interaction of several spatial and temporal following
pulses is scarcely determined yet. Thus, the aim of this study is to analyse the dynamic and interaction of two cavitation
bubbles by using high speed photography. The applied laser pulse energy, the energy ratio and the spot distance between
different cavitation bubbles were varied. Depending on a change of these parameters different kinds of interactions such
as a flattening and deformation of bubble shape or jet formation are observed. The effects will be discussed regarding the
medical ophthalmic application of fs-lasers. Based on these results a further research seems to be inevitable to
comprehend and optimize the cutting effect of ultra-short pulse laser systems with high (> 500 kHz) repetition rates.
Focussed femtosecond laser pulses are applied in ophthalmic tissues to create an optical breakdown and therefore a
tissue dissection through photodisruption. The threshold irradiance for the optical breakdown depends on the photon
density in the focal volume which can be influenced by the pulse energy, the size of the irradiated area (focus), and the
irradiation time. For an application in the posterior eye segment the aberrations of the anterior eye elements cause a
distortion of the wavefront and therefore an increased focal volume which reduces the photon density and thus raises the
required energy for surpassing the threshold irradiance. The influence of adaptive optics on lowering the pulse energy
required for photodisruption by refining a distorted focus was investigated. A reduction of the threshold energy can be
shown when using adaptive optics. The spatial confinement with adaptive optics furthermore raises the irradiance at
constant pulse energy. The lowered threshold energy allows for tissue dissection with reduced peripheral damage. This
offers the possibility for moving femtosecond laser surgery from corneal or lental applications in the anterior eye to
vitreal or retinal applications in the posterior eye.
Although femtosecond laser cell surgery is widely used for fundamental research in cell biology, the mechanisms in
the so-called low-density plasma regime are largely unknown. To date, it is still unclear on which time scales free
electron and free radical-induced chemical effects take place leading to intracellular ablation. In this paper, we
present our experimental study on the influence of laser parameters and staining on the ablation threshold. We
found that the ablation effect resulted from the accumulation of single-shot multiphoton-induced photochemical
effects finished within a few nanoseconds. In addition, fluorescence staining of subcellular structures significantly
decreased the ablation threshold. Based on our findings, we propose that dye molecules are the major source for
providing seed electrons for the ionization cascade.
We present a high-speed photographic analysis of the interaction of cavitation bubbles generated in two spatially separated regions by femtosecond laser-induced optical breakdown in water. Depending on the relative energies of the femtosecond laser pulses and their spatial separation, different kinds of interactions, such as a flattening and deformation of the bubbles, asymmetric water flows, and jet formation were observed. The results presented have a strong impact on understanding and optimizing the cutting effect of modern femtosecond lasers with high repetition rates (>1 MHz).
The delivery of extra cellular molecules into cells is essential for cell manipulation. For this purpose genetic materials
(DNA/RNA) or proteins have to overcome the impermeable cell membrane. To increase the delivery efficiency and cell
viability of common methods different nano- and micro material based approaches were applied. To manipulate the cells,
the membrane is in contact with the biocompatible material. Due to a field enhancement of the laser light at the material
and the resulting effect the cell membrane gets perforated and extracellular molecules can diffuse into the cytoplasm.
Membrane impermeable dyes, fluorescent labelled siRNA, as well as plasmid vectors encoded for GFP expression were
used as an indicator for successful perforation or transfection, respectively. Dependent on the used material, perforation
efficiencies over 90 % with a cell viability of about 80 % can be achieved. Additionally, we observed similar efficiencies
for siRNA transfection. Due to the larger molecule size and the essential transport of the DNA into the nucleus cells are
more difficult to transfect with GFP plasmid vectors. Proof of principle experiments show promising and adequate
efficiencies by applying micro materials for plasmid vector transfection. For all methods a weakly focused fs laser beam
is used to enable a high manipulation throughput for adherent and suspension cells. Furthermore, with these alternative
optical manipulation methods it is possible to perforate the membrane of sensitive cell types such as primary and stem
cells with a high viability.
The aim was to evaluate a method for visualizing fs laser pulse induced microincisions inside crystalline lens tissue. Porcine lenses were modified ex vivo by fs laser pulses to create defined planes at which lens fibers separate. Lens fiber orientation and fs laser-induced micro-incisions were examined using a confocal laser scanning microscope. Micro-incision visualization revealed different cutting effects depending on fs laser pulse energy, ranging from altered tissue scattering properties with all fibers intact to definite fiber separation with a wide gap. CLSM permits visualization and analysis and thereby control of fs laser pulse induced microincisions inside crystalline lens tissue.
In ophthalmology, femtosecond laser transections (photodisruption) in the vicinity of the retina need to be
performed with minimized threshold energy to not harm peripheral retinal tissue. However, the aberrations
of the anterior eye elements cause a distortion of the wavefront and therefore a raised threshold energy when
focussing into the posterior segment. We present an optical system that allows for correcting aberrations in eyes
using adaptive optics consisting of a deformable mirror and a Hartmann-Shack-Sensor with a novel light source.
If combined with femtosecond laser pulses this system offers the possibility for minimally invasive laser surgery
in the posterior eye segment.
Cross-linking of stromal collagen with Riboflavin and UVA radiation is an alternative treatment of keratoconus. After
the cross-linking a wound healing process starts with the regeneration of the abraded epithelial layer and the stromal
keratocyte-network. To clarify possible side effects by visualization we established an imaging platform for the
multimodal three-dimensional imaging of the cornea and looked for differences between normal and cross-linked rabbit
corneae. The microscopy system utilizes femtosecond laser light for two photon excitation of autofluorescent metabolic
compounds, second harmonic imaging in forward and backward direction for the study of stromal collagen-I structure
and confocal detection of the backscattered femtosecond laser light for cell detection. Preliminary results show
signatures of treatment 5 weeks after the intervention in all imaging modalities.
The resulting effects of the interaction between nanoparticles and laser irradiation are a current matter in research.
Depending on the laser parameters as well as the particles properties several effects may occur e.g. bubble formation,
melting, fragmentation or an optical breakdown at the surface of the nanoparticle. Besides the investigations of these
effects, we employed them to perforate the membrane of different cell lines and investigated nanoparticle mediated laser
cell perforation as an alternative optical transfection method. Therefore, the gold nanoparticles (GNP) of different shapes
were applied. Furthermore, we varied the methods for attaching GNP to the membrane, i.e. co-incubation of pure gold
nanoparticles and bioconjugation of the surface of GNP. The optimal incubation time and the location of the GNP at the
cell membrane were evaluated by multiphoton microscopy. If these GNP loaded cells are irradiated with a fs laser beam,
small areas of the membrane can be perforated. Following, extra cellular molecules such as membrane impermeable dyes
or foreign DNA (GFP vectors) are able to diffuse through the perforated area into the treated cells. We studied the
dependence of the laser fluence, GNP concentration, GNP size and shape for successful nanoparticle mediated laser cell
perforation. Due to a weak focusing of the laser beam a gentle cell treatment with high cell viabilities and high
perforation efficiencies can be achieved. A further advantage of this perforation technique is the high number of cells
that can be treated simultaneously. Additionally, we show applications of this method to primary and stem cells.
Light microscopy is one of the major tools in modern biology. The steady development of new microscopic
techniques leads to an correspondent improvement of biological methods. To expand the catalog of biological
experiments, we investigate the possibilities of optical projection tomography (OPT). This technique is based on
the already established X-Ray computed tomography. In contrast to most other three-dimensional microscopy
techniques it is able to create three dimensional data sets of the specimens natural absorption, staining and
fluorescence. Unfortunately, these advantages are opposed by a low resolution, reconstruction artifacts, and a
relatively big loss of fluorescence light. We reduced the disadvantage in resolution by applying physical filters in
the Fourier plane of the image path, which is not possible in X-Ray imaging yet.
The LASIK procedure is a well established laser based treatment in ophthalmology. Nowadays it includes a cutting of
the corneal tissue bases on ultra short pulses which are focused below the tissue surface to create an optical breakdown
and hence a dissection of the tissue. The energy of the laser pulse is absorbed by non-linear processes that result in an
expansion of a cavitation bubble and rupturing of the tissue. Due to a reduction of the duration of treatment the current
development of ultra short laser systems points to higher repetition rates. This in turn results in a probable interaction
between different cavitation bubbles of adjacent optical breakdowns. While the interaction of one single laser pulse with
biological tissue is analyzed reasonably well experimentally and theoretically, the interaction of several spatial and
temporal following pulses is scarcely determined yet. We present a high-speed photography analysis of cavitation bubble
interaction for two spatial separated laser-induced optical breakdowns varying the laser pulse energy as well as the
spatial distance. Depending on a change of these parameters different kinds of interactions such as a flattening and
deformation of bubble shape, asymmetric water streams and jet formation were observed. The results of this research can
be used to comprehend and optimize the cutting effect of ultra short pulse laser systems with high repetition rates
(> 1 MHz).
Biocompatibility studies of percutanous implants in animal models usually involve numerous lethal biopsies for
subsequent morphometric analysis of the implant-tissue interface. A common drawback of the study protocol is
the restriction of the analysis to one final time point. In this study optical coherence tomography (OCT) was used
to visualize and enable quantification of the local skin anatomy in the vicinity of a percutaneous implant in an
animal model using hairless mice. Non invasive in vivo optical biopsies were taken on predetermined time points
after implantation and ex vivo in situ at the day of noticeable inflammation. The custom made Fourier-domain
OCT system was programmed for imaging with different scanning schemes. A spoke-pattern of 72 cross-sectional
scans which was centred at the midpoint of the circular shaped implants was acquired and worked best for the
in-vivo situation. Motion-artefact-free three-dimensional tomograms were obtained from the implant site before
excision and preparation for histology. Morphometric parameters such as epithelial downgrowth, distance
to normal growth and tissue thickness were extracted from the images with a simple segmentation algorithm.
Qualitatively, the OCT B-Scans are in good agreement with histological sections. Therefore, OCT can provide
additional valuable information about the implant-tissue interface at freely selectable time points before the
lethal biopsy. Locally confined quantitative assessments of tissue-implant interaction for in vivo postoperative
monitoring can be carried out.
Lately, several groups successfully used ultrashort laser pulses to selectively permeabilize the membrane of living cells
to achieve transport of foreign molecules, like DNA, into the cells. For this, the high field intensities of tightly focused
laser pulses are used to induce multiphoton absorption and the creation of a small scale optical breakdown at the
membrane of the target cell. Afterwards, DNA or other foreign molecules are able to diffuse into the cell and achieve,
for example, transfection of living cells. However, the cell throughput of this method is low, as, due to tight focusing.
We present a technique to achieve fs-laser transfection in living cells at higher throughput by implementing optical traps
into microfluidic chips. For this, a trapping laser beam, is coupled into a microscope setup and combined with a Ti:Sa fslaser
beam to achieve simultaneous trapping and optical perforation.
Femtosecond (fs) laser-based cell surgery is typically done in two different regimes, at kHz or MHz repetition rate. Formation of reactive oxygen species (ROS) is an often predicted effect due to illumination with short laser pulses in biological tissue. We present our study on ROS formation in single cells in response to irradiation with fs laser pulses depending on the repetition rate while focusing into the cell nucleus. We observed a significant increase of ROS concentration directly after manipulation followed by a decrease in both regimes at kHz and MHz repetition rate. In addition, effects of consecutive exposures at MHz and kHz repetition rate and vice versa on ROS production were studied. Irradiation with a MHz pulse train followed by a kHz pulse train resulted in a significantly higher increase of ROS concentration than in the reversed case and often caused cell death. In the presence of the antioxidant ascorbic acid, accumulation of ROS and cell death were strongly reduced. Therefore, addition of antioxidants during fs laser-based cell surgery experiments could be advantageous in terms of suppressing photochemical damage to the cell.
Presbyopia is an age related effect which affects every human at the age of about 40 years. So far reading glasses
are the conventional treatment. According to Helmholtz' theory of accommodation one of the mayor reasons for
the development of presbyopia is the increasing sclerosis of the lens. In contrast to that the ciliary muscle and
the lens capsule remain mostly active and elastic the whole life. So a possible treatment could be the increase of
the flexibility of the lens by creating gliding planes with fs-laser pulses inside the lens tissue.
In former studies it was shown that fs-laser pulses were able to increase the flexibility of ex vivo porcine lenses
as well as ex vivo human donor lenses. Our current aim was to evaluate the effect of the fs-laser pulses on the
crystalline lens of living rabbit eyes due to the fs-lentotomy treatment. The main focus of the evaluation was
the exclusion of possible side effects of the treatment like cataract formation or retina damage. The treated eyes
were monitored using optical coherence tomography (OCT) and Scheimpflug imaging for localizing and studying
the tissue effects of the incisions. Furthermore histological sections of the lens and retina were prepared. The
rabbits were investigated pre operatively and up to six months post operatively.
The fs-laser induced micro incisions were successfully applied to the left lens of each rabbit. The micro
incisions within the crystalline lens were detectable with OCT and Scheimpflug imaging up to six month. The
imaging within the lens showed a progressive fading of the incisional opacities generated by the femtosecond
laser during the six months and no indication of cataract formation was found. OCT and Scheimpflug images
emphasize themselves as necessary tools to monitor the micro incisions over time. Histopathological sections of
the lens tissue support the findings of the non invasive imaging techniques. Also the histopathological sections
of the retina show no thermal induced change due to the irradiation of the fs-pulses.
We present our results for nanoparticle mediated laser
poration as an alternative transfection technique. As a fundamental part for the perforation of the cell membrane the interactions of gold nanoparticles and living cells were studied.
A prominent laser based treatment in ophthalmology is the LASIK procedure which nowadays includes a cutting of the
corneal tissue based on ultra short pulses. Focusing an ultra short laser pulse below the surface of biological tissue an
optical breakdown is caused and hence a dissection is obtained. The laser energy of the laser pulses is absorbed by nonlinear
processes. As a result a cavitation bubble expands and ruptures the tissue. Hence positioning of several optical
breakdowns side by side generates an incision. Due to a reduction of the duration of the treatment the current
development of ultra short laser systems points to higher repetition rates in the range of hundreds of KHz or even MHz
instead of tens of kHz. This in turn results in a probable occurrence of interaction between different optical breakdowns
and respectively cavitation bubbles of adjacent optical breakdowns. While the interaction of one single laser pulse with
biological tissue is analyzed reasonably well experimentally and theoretically, the interaction of several spatial and
temporal following pulses is scarcely determined yet. Thus the aim of this study is to analyse the dynamic and interaction
of two cavitation bubbles by using high speed photography. The applied laser pulse energy, the energy ratio and the spot
distance between different cavitation bubbles were varied. Depending on a change of these parameters different kinds of
interactions such as a flattening and deformation of bubble shape or jet formation are observed. Based on these results a
further research seems to be inevitable to comprehend and optimize the cutting effect of ultra short pulse laser systems
with high (> 1 MHz) repetition rates.
We developed a laryngoscope with an integrated OCT beam path for office-based non-contact imaging of human
vocal folds. In combination with conventional videolaryngoscopy superficial and subsurface lesions can be detected.
For error-free interpretation of OCT images obtained in office-based examination motion artifacts have
to be considered. To demonstrate the implications on OCT images we simulated probe and patient movements
for different commercial systems representing the three OCT modalities and analyzed the OCT data. Our results
show that time domain and fourier domain OCT with a swept light source are probably better suited for noncontact
imaging of awake patients than the current generation of fourier domain OCT engines with spectrometer
design.
The hearing performance with conventional hearing aids and cochlear implants is dramatically reduced in noisy environments and for sounds more complex than speech (e. g. music), partially due to the lack of localized sensorineural activation across different frequency regions with these devices. Laser light can be focused in a controlled manner and may provide more localized activation of the inner ear, the cochlea. We sought to assess whether visible light with parameters that could induce an optoacoustic effect (532 nm, 10-ns pulses) would activate the cochlea. Auditory brainstem responses (ABRs) were recorded preoperatively in anesthetized guinea pigs to confirm normal hearing. After opening the bulla, a 50-µm core-diameter optical fiber was positioned in the round window niche and directed toward the basilar membrane. Optically induced ABRs (OABRs), similar in shape to those of acoustic stimulation, were elicited with single pulses. The OABR peaks increased with energy level (0.6 to 23 µJ/pulse) and remained consistent even after 30 minutes of continuous stimulation at 13 µJ, indicating minimal or no stimulation-induced damage within the cochlea. Our findings demonstrate that visible light can effectively and reliably activate the cochlea without any apparent damage. Further studies are in progress to investigate the frequency-specific nature and mechanism of green light cochlear activation.
Fs-lasers are widely used for microsurgery and micromachining. Due to nonlinear interaction of ultrashort pulses with tissue or matter precisions of a few μm can be achieved. But particularly in the field of surgery this precision can not be obtained as the devices for diagnostics and treatment have to be changed due to separate systems. We show a combined system of a fs-laser and a Fourier-Domain optical coherence tomography (FD-OCT) enabling to cut and image the region of interest alternately. The FD-OCT offers non-invasive imaging at an axial resolution of 6, 2 μm and a transverse resolution of 3 μm in air which is comparable to the interaction zone of the fslaser-pulses. OCT-aided subsurface cutting is successfully demonstrated on biological ex-vito samples of porcine
cornea and larynx. Furthermore it appeared that in situ OCT imaging enables to monitor cuts produced with pulse energies close to the energy threshold. In conclusion, this setup demonstrates the potential of a system combining cutting and OCT imaging within the same optical setup without the need of changing devices.
The success of conventional hearing aids and electrical cochlear implants have generally been limited to hearing in quiet situations, in part due to a lack of localized (i.e., frequency specificity) sensorineural activation and subsequent impaired speech discrimination in noise. Laser light is a source of energy that can be focused in a controlled manner and may provide more localized activation of the inner ear, the cochlea. Compound action potentials have been elicited using 2.12 µm laser pulses through activation of auditory nerve fibers (Izzo et al. 2006). Laser stimulation (813 nm) of the cochlea has shown to induce basilar membrane motion and cochlear microphonic potentials (Fridberger et al. 2006). We sought to assess if visible light (green, 532 nm, 10 ns pulses) could be used to consistently activate the cochlea. The laser parameters were selected based on our initial attempt to induce an optoacoustic effect as the energy transfer mechanism to the cochlea. Click evoked auditory brainstem responses (AABRs) were recorded preoperatively in ketamine-anesthetized guinea pigs to confirm normal hearing. The bulla and then the cochlea were exposed. Optically evoked ABRs (OABR) were recorded in response to laser stimulation with a 50 µm optical fiber (532 nm, 10 ns pulses, 500 repetitions, 10 pulses/s; Nd:YAG laser) at the round window (RW) directed towards the basilar membrane (BM).
OABRs similar in morphology to acoustically evoked ABRs, except for shorter latencies, were obtained for stimulation through the RW with energy levels between 1.7-30 µJ/pulse. The OABRs increased with increasing energy level reaching a saturation level around 13-15 µJ/pulse. Furthermore the responses remained consistent across stimulation over time, including stimulation at 13 µJ/pulse for over 30 minutes, indicating minimal or no damage within the cochlea with this type of laser stimulation.
Overall we have demonstrated that laser light stimulation with 532 nm has potential for a new type of auditory prosthesis that can activate the cochlea without any apparent functional damage. Further studies are needed to determine the optimal laser parameters and fiber placement locations for localized and tonotopic activation.
Investigation on the interaction of small particles, e.g. gold nanoparticles with light is a current field of high interest. As
light can be absorbed, enhanced or scattered by the nanoparticles a wide variety of possible applications become
possible. If the electrons of such a nanoparticles oscillate with the incident light, plasmon resonances occur. Provided
that these particles are brought very close to a cell, the cell membrane gets perforated due to the laser induced effect. We
investigate nanoparticle mediated laser perforation as an alternative technique for cell transfection. By using weakly
focussed femtosecond laser pulses, 150 nm gold particles were stimulated to perforate the cell membrane. Through the
perforated area of the membrane macromolecules e.g. DNA are able to enter the cell. By this technique GFSHR-17 rat
cells were successfully transfected with GFP vector and the dependence on laser parameters and concentration were
studied. Even after 48 hours after manipulation the transfected cells show no indications of apoptosis or necrosis. This
technique allows the transfection of cells by opto-perforation without the need of tight focusing conditions and single
cell targeting- opening the way for a wide field of applications.
The manipulation of cells by femtosecond (fs) laser pulses became a very important tool in cell biology. In terms
of learning more about the function of the cell compartments and the cell dynamics, single cell organelles are
manipulated by laser pulses. Meanwhile the cell reaction is observed by different microscopy methods. The
parameters of the laser irradiation have to be chosen carefully to minimize unwanted side effects during the
treatment and to prevent cell damage or cell death. In many applications, it is not known what happens due to
the laser irradiation on the molecular level. The formation of reactive oxygen species (ROS) is an often predicted
effect due to photo disruption in biologic tissue. In this paper, we present our study of the ROS formation during
the irradiation of fs laser pulses for disruption of single cell organelles. The quantity of ROS formation depends
strongly on the pulse energy of the laser. Therefore the creation of ROS was additionally studied while scanning
the laser at low energy for multiphoton microscopy.
Due to nonlinear interaction with optical transparent and scattering samples the femtosecond technology is a
very useful tool for high precision micro surgery on biological tissues. At the same time femtosecond lasers are
ideal light sources for imaging methods such as optical coherence tomography (OCT) due to the broad spectrum
of the laser, which is necessary for creating ultra short pulses. Using OCT structures within biological tissues
can be imaged non invasive with a resolution within the low m-range.
The combined use of an ultra short pulse laser for cutting of biological tissues as well as imaging via OCT is a
very interesting tool. It opens up a wide range of new surgery techniques and improves many existing methods
due to high precision and high flexibility of the cutting process.
Therefore we combined a femtosecond cutting system and a fourier domain OCT. In this attempt the OCT is
operated with an SLD and is used alternately to the cutting system. The OCT is integrated into the optical
path which enables in situ imaging of the surgery area before and after treatment.
Ultrashort laser pulses recently found extensive application in micro- and nanostructuring, in refractive surgery
of the eye, and in biophotonics. Due to the high laser intensity required to induce optical breakdown, nonlinear
plasma formation is generally accompanied by a number of undesired nonlinear side-effects such as self-focusing,
filamentation and plasma-defocusing, seriously limiting achievable precision and reproducibility. To reduce pulse
energy, enhance precision, and limit nonlinear side effects, applications of ultrashort pulses have recently evolved
towards tight focusing using high numerical aperture microscope objectives. However, from the theoretical and
numerical point of view generation of optical breakdown at high numerical aperture focusing was barely studied.
To simulate the interaction of ultrashort laser pulses with transparent materials, a comprehensive numerical
model taking into account nonlinear propagation, plasma generation as well as the pulse's interaction with
the generated plasma is introduced. By omitting the widely used scalar and paraxial approximations a novel
nonlinear propagation equation is derived, especially suited to meet the conditions of high numerical aperture
focusing. The multiple rate equation (MRE) model is used to simultaneously calculate the generation of free
electrons. Nonparaxial and vectorial diffraction theory provides initial conditions.
The theoretical model derived is applied to numerically study the generation of optical breakdown plasmas,
concentrating on parameters usually found in experimental applications of cell surgery. Water is used as a model
substance for biological soft tissue and cellular constituents. For focusing conditions of numerical aperture
NA < 0.9 generation of optical breakdown is shown to be strongly influenced by plasma defocusing, resulting in
spatially distorted breakdown plasmas of expanded size. For focusing conditions of numerical aperture NA ≥ 0.9
on the other hand generation of optical breakdown is found to be almost unaffected by distortive side-effects,
perfectly suited for material manipulation of highest precision.
Up to now reading glasses are the conventional treatment of presbyopia, an age related effect for every human.
According to the Helmholtz theory the reason for the development of accommodative loss is a decreasing elasticity
of the lens due to the increasing sclerosis. Since the ciliary muscle and the lens capsule remain active and elastic
the whole life, a possible treatment could be the increase of the flexibility by creating gliding planes with fs-laser
pulses.
flexibility of ex vivo porcine as well
as human donor lenses with a laboratory laser system. We will present new results with a compact 100 kHz
repetition rate turn key laser system which speeds up the treatment time by a factor of 10. This will offer the
opportunity for future clinical trials. Furthermore first in-vivo results on rabbits are presented.
Optical coherence tomography (OCT) is a new promising imaging modality in laryngology as yet of undetermined
value. The objective of the present study was to evaluate the ability of this noninvasive method to make a reliable
prediction of diagnosis in laryngeal disease. In a prospective study, 225 benign and malignant laryngeal lesions were
examined with a fiber-based OCT system in contact mode during elective microlaryngoscopy. Intraoperative OCT
findings were compared to conventional histopathology supplying a correct specific diagnosis in about 90% of cases.
With increasing experience, a reliable prediction of invasive tumor growth and often the exact grade of dysplasia were
possible. Due to the current spatial resolution of OCT, microinvasive cancer could not be safely delineated from severe
dysplasia or carcinoma in situ. In contrast, benign lesions made no essential diagnostic difficulties due to their characteristic
OCT image. In conclusion, OCT has proved to be very useful in the diagnostic investigation and the intraoperative
monitoring of laryngeal disease.
Surgery of benign pathological alterations of the vocal folds results in permanent disphonia if the bounderies of the vocal fold layers are disregarded. Precise cutting with a femtosecond laser (fs-laser) combined with simultanous imaging of the layered structure enables accurate resections with respect to the layer boundaries. Earlier works demonstrated the capability of optical coherence tomography (OCT) for utilization on vocal folds. The layered structure can be imaged with a spatial resolution of 10-20&mgr;m up to a depth of 1.5mm. The performance of fs-laser cutting was analyzed on extracted porcine vocal folds with OCT monitoring. Histopathological sections of the same processed samples could be well correlated with the OCT images. With adequate laser parameters thermal effects induced only negligable damage to the processed tissue. The dimensions of the thermal necrosis were determined to be smaller than 1&mgr;m. OCT contolled fs-laser cutting of porcine vocal fold tissue in the &mgr;m range with minimal tissue damage is presented.
According to Helmholtz' theory of accommodation one of the mayor reasons for the development of presbyopia is
the increasing sclerosis of the lens. One concept to overcome this hardening of the lens is to regain its flexibility
by inducing gliding planes inside the lens. Femtosecond laser pulses are a suitable tool for this treatment.
Showing in former work that we could increase the flexibility of enucleated porcine (ex vivo) lenses up to 25%,
we focused our recent work on human autopsy lenses. The age of the human donors ranged between 20 and
70 years. For an evaluation of the gain in flexibility the lens' thickness was measured undertaking the Fisher's
spinning test before and after laser treatment. Depending on the age and the quality of applied cutting pattern
the lens thickness increased after treatment up to 0.4 mm leading to an theoretical increase of several dioptres
of optical power. The flexibility could be increased up to 70 % compared to the measurements before treatment.
Since the age of the human donors had a broad range, leading to different degrees of lens hardening, the variance
of the measured flexibility changes was up to 30%. An addition the influence of the laser treatment onto the lens
on the accommodation amplitude will be shown in a three dimensional finite-element simulation.
Due to nonlinear interaction with optical transparent samples the femtosecond technology is a very useful tool
for high precision micro surgery on biological tissues. At the same time femtosecond lasers are ideal light sources
for imaging methods such as optical coherence tomography (OCT) due to the broad spectrum of the laser, which
is necessary for creating ultra short pulses. Using OCT structures within biological tissues can be imaged non
invasive with a resolution within the low μm-range. The combined use of an ultra short pulse laser for cutting of
biological tissues as well as imaging via OCT is a very interesting tool. It opens up a wide range of new surgery
techniques and improves many existing methods due to high precision and high flexibility of the cutting process.
Therefore we combined a femtosecond cutting system and a fourier domain OCT. In a first attempt the OCT is
driven with an SLD and is used alternately to the cutting system. The OCT is integrated into the optical path
which enables in situ imaging of the surgery area.
Optical coherence tomography (OCT) is a new non-invasive method to investigate biological tissue. It is particularly
suitable for examination of human vocal folds due to its optical penetration depth of around 1.5mm.
We developed a modified laryngoscope with an integrated OCT beam path for non-contact imaging of human
vocal folds. In vivo studies on awake patients show that synchronous OCT and conventional laryngoscopy
works well. For enabling the choice of the appropriate OCT encoding technique for OCT-based laryngoscopy
measurements on simulated vibrations were performed with time domain (TD) and fourier domain (FD) OCT
devices. The results show that TD-OCT is much more suitable for non-contact imaging than FD-OCT.
In studies on porcine and monkey vocal folds the performance of femtosecond laser tissue ablation was
analyzed with OCT monitoring. Histopathological sections could be well correlated with the OCT images. In
future applications a combined system that uses one femtosecond laser as light source for tissue ablation as well
as for OCT imaging is conceivable.
In conclusion, OCT can be used as a pre-, intra- and post-operative diagnostic instrument for analysing
the vocal fold structure down to the vocalis muscle allowing a more precise indication for potential subsequent
invasive procedures. We suggest that OCT should be used in addition to established examination methods for
diagnostic evaluation of vocal fold malignancies and functional alterations.
Ultrashort laser pulses tightly focused provide intensity sufficient to initialize nonlinear ionization processes. Thus a plasma is generated in the focal region eventually resulting in optical breakdown. The deterministic character of this nonlinear interaction enables the generation of precise and highly reproducible material alteration. To gain better spatial precision applications have recently evolved strongly towards tight focusing of ultrashort pulses using microscope objectives as focusing units. The pulse energy required to generate optical breakdown was thus reduced to nanojoules or even below. The mechanical effects subsequent to plasma generation can be minimized to the very focus. Cell surgery with ultrashort pulses enables to precisely ablate cell organelles without observable hazardous effects to the surroundings or the entire cell.
To numerically investigate the nonlinear interaction of ultrashort pulses with transparent media, a model including both nonlinear pulse propagation and plasma generation is introduced. The numerical code is based on a (3+1)-dimensional nonlinear Schrödinger equation describing the pulse propagation and the interaction with the density of free electrons that are generated in the focus. The nonlinear wave equation was derived taking into account both nonparaxial and vectorial effects to accurately include tight focusing at high numerical aperture. A multi rate equation model for dielectrics recently published by B. Rethfeld is used to simultaneously calculate the generation of free electrons.
Numerical calculations based on this model are used to understand the dependence between size, geometry and density of optical breakdown plasmas in various focusing geometries of high numerical aperture. The code enables to use arbitrary initial conditions for the laser field in the focus. At high numerical aperture it is most important to start the simulation using realistic initial conditions. Especially the vectorial character of the electric field is most important to be considered. Thus a vectorial diffraction integral was used to calculate initial conditions at high numerical aperture.
The code is applicable to any transparent Kerr medium, whose linear and nonlinear optical parameters are known. Within this work the code was applied to water as a model substance to biological soft tissue and cellular constituents.
The laser microtome (LMT) is designed to slice biological tissue and various materials with high precision. The
cutting process is performed by an ultrafast laser, emitting infrared light with high repetition rate of about 10
MHz. Biological Tissue can be sliced without fixation and embedding. Therefore cutting of native tissue is
possible. Due to the non contact procedure further investigations of the material like immuno histological tests
can be performed. At present slices with a thickness of 5 - 100&mgr;m in different biological tissues like cartilage,
kidney, lung and cornea has been shown. In conjunction with a 3D imaging system like optical coherence
tomography, preparation of 3D tissue volumes is possible too.
Presbyopia is one age related effect every human is suffering beginning at the age of about 45 years. Reading
glasses are the conventional treatment so far. According to the Helmholtz theory the loss of accommodation in
age is due to the hardening and the resulting loss of elasticity of the crystalline lens. However the ciliary muscle
and the lens capsule stay active, respectively. Therefore a possible treatment concept is to regain the flexibility
by inducing gliding planes in form of microcuts inside the lens. The increase of flexibility in young porcine lenses
by different cutting patterns was shown by Ripken et al.1, 2 who verified the increase in flexibility by the spinning
test introduced by Fisher.3
We will present our first measurements of flexibility increase of human donor lenses. Furthermore the influence
of the laser cuts into the lens on the accommodation amplitude will be shown in a three dimensional finite-element simulation.
Fluorescence imaging of cells and cell organelles requires labeling by fluorophores. The labeling of living cells is often done by transfection of
fluorescent proteins. Viral vectors are transferring the DNA into the cell. To avoid the use of viruses, it is possible to perforate the cell
membrane for example by electro-shocks, the so called electroporation, so that the fluorescent proteins can diffuse into the cell. This method
causes cell death in up to 50% of the treated cells because the damage of the outer membrane is too large.
A less lethal perforation of the cell membrane with high efficiency can be realized by femtosecond (fs) laser pulses. Transient pores are created by focusing the laser beam for some milliseconds on the membrane. Through this pore, the proteins can enter into the cell. This was demonstrated in a proof of principle experiment for a few cells, but it is essential to develop an opto-perforation system for
large numbers of cells in order to obtain statistically significant samples for biological experiments. The relationship between pulse energy,
irradiation time, repetition rate and efficacy of the transfer of a chromophor into the cells as well as the viability of the cells was analysed. The cell viability was observed up to 90 minutes after manipulation.
The most probable reason for presbyopia is an age related loss of elasticity of the lens. It progresses typically during the whole life and at the age of about 45 it leads to a considerable loss of the ability to accommodate within the next decade. However, both, the ciliary muscle and the lens capsule stay active and elastic, respectively. With respect to this, one concept is to regain the deformability of the lens without changing the capsule or zonular apparatus. Since the investigations of Ripken et al. proofed that the flexibility of the presbyopic lens tissue can be increased through the creation of fs-laser induced microcuts inside the lens, this is one possible approach to treat presbyopia. On this account a finite-element-method model with ANSYS of the human lens during accommodation will be presented. The analysis premises all lens materials to be linear elastic and allow large displacements. A first analysis of this method for the treatment of presbyopia is accomplished. Therefore the mechanical analysis of untreated and treated lens are compared. In addition ex-vivo elasticity measurements of untreated and treated lenses will be presented. As a result an improvement of the flexibility of the lens tissue is found and as its consequence a change of the lens radii of curvature is established. After suitable processing of the output data the change in optical power between untreated and treated lenses are calculated. The finite element simulation shows similar behaviour compared to the treated porcine lenses.
Rapid development of new laser technologies enabled the application of ultra short lasers in refractive surgery. Focused ultra short laser pulses in near-infrared spectral range can generate a laser induced breakdown (LIB) in the cornea, which will disrupt the tissue. Cutting depth and position can be established by varying the laser focus. The fs-LASIK technique allows both flap and lenticule to be formed by using fs-pulses without the presence of any mechanical impact. During the cutting process not all of the pulse energy is deposited into the cornea; approximately half of the remaining energy propagates through the eye and reaches the retina. Though defocused, the transmitted energy can still induce damage to the retina due to absorption by the retinal pigment epithelium and the transfer of thermal energy to surrounding tissue. The fs-LASIK process was simulated with two laser systems; one continous-wave and one in the fs-regime. For the simulation the exposure time and focusing numerical aperature which defines the retinal spot size were varied. The Damage thresholds of the laser beam exposed eyes were determined in terms of ophthalmoscopic and histopathologic observations.
Recently ultrashort laser pulses became most important for micro structuring and biomedical applications such as refractive surgery. Ultrashort laser pulses tightly focused to a small spot easily provide intensity sufficient to induce nonlinear ionization. A plasma is generated and heated in the focus resulting in optical breakdown. The energy deposited in the plasma and the mechanical effects subsequent to optical breakdown are utilized by modern applications of ultrashort laser pulses to induce controlled highly reproducible material alteration. A model including both nonlinear pulse propagation and plasma generation is introduced to numerically investigate the interaction of ultrashort laser pulses with the self-induced plasma in the vicinity of the focus. The numerical code is based on a (3+1)-dimensional nonlinear Schroedinger equation describing the pulse propagation. A multi rate equation model recently published by B. Rethfeld is used to simultaneously calculate the generation of free electrons. It is the first numerically simple approach to describe nonlinear ionization that allows a non static energy distribution of free electrons in the conduction band. The code is applicable to any transparent Kerr medium, whose linear and nonlinear optical parameters are known. Numerical calculations based on this model are used to understand the dependence between the size, the geometry and the free electron density of ultrashort laser pulse induced optical breakdown plasmas in various focusing geometries. The code enables to use arbitrary initial conditions for the laser field in the focus. More realistic focusing scenarios than the simple assumption of focused gaussian beams can be taken into account. Nonlinear side effects, such as streak formation occurring in addition to optical breakdown during ultrashort pulse refractive eye surgeries can be numerically investigated.
Multiphoton microscopy is a very promising method for 3D imaging
of living cells. The fluorochromes are solely excited at the laser
focus by multiphoton absorption using near-infrared femtosecond
laser pulses. The arising fluorescence serves for a pixel-to-pixel
imaging with a resolution in the submicron range. At higher laser
powers, the multiphoton absorption creates a micro plasma which
induces an outwardly propagating shock wave. The rapidly expanding
cavitation bubble causes disruption of the material, with hardly
any interaction with the surrounding tissue as the optical breakdown proceeds faster than the thermal conduction.
This combination offers the possibility of simultaneous manipulation and analysis of living cells or cell organelles. Manipulation is achieved using laser pulses with an energy of a few nanojoules while imaging is done at less than 1 nJ. The obtained resolution allows the precise cutting of single cell organelles without compromising the cells` viability. Thus, the implementation is excellently suited for cell surgery. We conducted ablation of different subcellular structures, like mitochondria, at different pulse energies within living cells while studying cell viability.
The current standard procedure to ensure the diagnosis, if tissue is malignant, is still an invasive one. Optical coherence tomography (OCT) is a new non-invasive method to investigate biological tissue. In this study OCT was used on porcine and on human vocal folds. The optical penetration depth of the used radiation is up to 2 mm.
Three different OCT application systems were used. The first is a high resolution OCT, which works in contact mode. It was used to examine porcine vocal folds ex vivo. Porcine vocal folds were assigned to defined areas and examined by OCT in contact mode followed by traditional histo-morphological analysis. The second OCT is fiber based. It also works in contact mode. Images of human vocal folds were done in contact mode. They were compared with a typical histo-morphological image of a human vocal fold. The third application system works in non contact to the tissue. It was integrated in a conventional laryngoscope. Human vocal folds were examined in vivo. Single layers of the vocal folds could be distinguished from each other with all used systems. Pathological alterations could be seen. Imaging is possible in real time. General anaesthesia is not necessary. OCT makes it possible to get a view under the surface of the vocal fold without being invasive.
A finite-element-method model with ANSYS 8.0 of a 29 year old human lens during accommodation will be presented.
The required data, to draw and calculate a two dimensional, axis-symmetric model of the human lens is inherited from
various sources. Furthermore the analysis premises all lens materials to be linear elastic and allows large displacements.
A first analysis of a possible method for the treatment of presbyopia by fs-laser induced microcuts is accomplished.
Therefore a mechanical analysis of an untreated and a treated lens are compared. As a result an improvement of the
flexibility of the lens tissue is found and as its consequence a change of the lens' radii of curvature is established. After
a suitable processing of the output data a linear Gaussian ray trace is performed and a minor change in the optical power
between the untreated and treaded human lens is perceived. By calculation of the discrete optical power of the anterior
and posterior surface on the one hand and the overall optical power on the other hand an interpretation of the
effectiveness resulting from the treatment is offered. It is ascertained that the change in optical power of the anterior lens
surface is increased while the optical power of the posterior lens surface is decreased, almost compensating each other.
A possible explanation for this phenomenon is given and a suggestion of how to increase the effectiveness of the
treatment is discussed.
Optical coherence tomography (OCT) is a new non-invasive method to investigate biological tissue with a penetration depth of up to three millimeters. In this study OCT was used on the porcine larynx. The vocal folds were assigned to defined areas and examined by OCT followed by traditional histo-morphological analysis.
We were able to validate this new method by showing that both the OCT-tomographs and the histological assays showed a clear demarcation of epithelium mucosae from the deeper layers.
The current standard procedure to ensure the diagnosis, if tissue is malignant, is still an invasive one. Our results show that OCT allows the collection of crucial information about the quality of morphological changes on a vocal fold without the necessity of being invasive. Thus, we propose that optical coherence tomography should be introduced as a new and easy method for the detection of morphological changes of the vocal fold complementary to other established methods.
According to Helmholtz' theory of accommodation one of the mayor reasons for the development of presbyopia is the increasing sclerosis of the lens. One concept to delay the process of sclerosis or even regain the deformation ability of the lens might be the treatment of the lens by femtosecond laser pulses. Our aim was to evaluate appropriate laser parameters for this possible treatment and to analyse potential changes in deformation ability of the treated lenses. We performed different cutting patterns in enucleated pig lenses (ex vivo) using the disruptive effect of an ultrafast near-infrared laser induced optical breakdown. Pulse energies and spot separation of the laser pulses were varied to investigate the effect on the generated cut. For an evaluation of the gain in deformation ability the lenses were rotated before and after treatment and the changes in lens thickness due to centrifugal forces were measured. In result, a smooth cutting was possible with appropriate parameters. The experiments showed an increase of elasticity in 70% of the eyes. When the lenses were treated more statistically, an average deformation ability increase of nearly 20%, determined by the change of thickness between untreated and treated lens, was measured.
Ultrashort laser pulses are increasingly used in refractive eye
surgery to cut inside transparent corneal tissue. This is
exploited by the fs-LASIK procedure which affords the opportunity
to correct ametropia without any mechanical effects. The cutting
process is caused by the optical breakdown occurring in the laser
focus. During this process only a certain amount of the pulse
energy is deposited into the tissue. The remaining pulse energy
propagates further through the eye and interacts with the retina
and the strong absorbing tissue layers behind. Therefore this
investigation shall clarify if the intensity of the remaining
laser pulse and the resulting temperature field can damage the
retina and the surrounding tissue. Threshold values of the retinal
tissue and theoretical calculations of the temperature field will
be presented.
In the focal region of tightly focused ultrashort laser pulses, sufficient high intensities to initialize nonlinear ionization processes are easily achieved. Due to these nonlinear ionization processes, mainly multiphoton ionization and cascade ionization,
free electrons are generated in the focus resulting in optical breakdown. A model including both nonlinear pulse propagation and plasma generation is used to calculate numerically the interaction of ultrashort pulses with their self-induced plasma in the vicinity of the focus. The model is based on a (3+1)-dimensional nonlinear Schroedinger equation describing the pulse propagation coupled to a system of rate equations covering the generation of free electrons.
It is applicable to any transparent Kerr medium, whose linear and nonlinear optical parameters are known. Numerical calculations based on this model are used to understand nonlinear side effects, such as streak formation, occurring in addition to optical breakdown during short pulse refractive eye surgeries like fs-LASIK. Since the optical parameters of water are a good first-order approximation to those of corneal tissue, water is used as model substance. The free electron density distribution induced by focused ultrashort pulses as well as the pulses spatio-temporal behavior are studied in the low-power regime around the critical power for self-focusing.
The most probable reason for presbyopia is an age-related loss of
the elasticity of the lens. It develops through the whole life,
but is first noticeable typically at the age of about 45. From that on it leads within 15 years to a total loss of the accommodation ability. However, both, the ciliary muscle and the lens capsule
stay active and elastic, respectively. With respect to this, a
possible treatment conception is to increase or regain the elasticity. The possibility to increase elasticity with ps-laser induced cuts inside the lens was already shown by Krueger. We made an improvement in cutting quality while using a fs laser with 5~kHz repetition rate emitting in the near infrared. Different fs-laser-induced μm smooth cuts inside fresh enucleated ex-vivo pig lenses will be presented.
When ultrashort laser pulses are focused inside transparent materials, extremely high field intensities can easily be achieved in the focal volume leading to nonlinear interaction with the material. In corneal tissue this nonlinear interaction results in an optical breakdown that may serve as a cutting mechanism in ophthalmology. As a side effect of optical breakdown in corneal tissue, streak-like structures have been observed as discoloration
in histological sections under a light microscope. To investigate the streak formation, a numerical model including nonlinear pulse propagation due to self-focusing, group velocity dispersion, and plasma defocusing due to generated free electrons is presented.
The model consists of a (3+1)-dimensional nonlinear Schroedinger equation, describing the pulse propagation coupled to an evolution equation covering the generation of free electrons. The rate equation contains multi photon ionization as well as avalanche ionization. The model is applicable to any transparent Kerr-medium.
Creating individual complex three dimensional structures in HIP-zirconia by conventional mechanical machining, e.g. milling, is time consuming and subject to significant loss in bending strength due to microcracking during the milling process. Utilizing ultra-short laser pulses, individual complex three dimensional microstructures can be created very precisely without significant damage to the structure. This advantage is used to process HIP-zirconia in order to create dental restorations. To evaluate efficiency and quality, different laser parameters such as pulse duration, pulse energy and ablation strategies were studied. The maximum ablation rate was found at 400 fs.
ELAN is a new method for treating atherosclerotic vessels. Its purpose is to restore wall flexibility by removing arterial wall tissue from the outer arterial layer. This leads to expansion of the narrowed vessel resulting in increased blood flow. We present results of treatment of arteries in vitro. Tissue removal was done by excimer-laser ablation using ns-pulses of 193nm wavelength. We also discuss therapy control. OCT images and light diffusion measurements are presented.
The most probable reason for presbyopia, the age-related loss of the
accomodation-ability of the eye, is an age-related loss of the elasticity of the lens. To increase the elasticity, resp. regain elasticity we performed different fs-laser-induced cuts with an near-infrared 5 kHz femtosecond laser inside ex-vivo and in-vivo rabbit lenses. Sagittal and annulus cutting patterns in the lens were produced by focusing the laser beam through the cornea inside the lens and creating a laser induced optical breakdown. The cutting results were recorded with light microscopy and finally, after euthanization, changes in the optical fibers of the lens tissue were described.
Ultrafast lasers are suitable for different surgical procedures in the eye: As a main goal in our group, the application of fs-lasers in refractive surgery is studied. Therefore,experiments in living rabbits were conducted, with wound healing studies up to 120 ays after surgery. Moreover,highly precise cuts inside the cornea were used to do keratoplasty an lamellar keratoplasty. Further applications like a new technique to overcome presbyopia are shown.
ELAN is a new method for treating atherosclerotic vessels. Its purpose is to restore wall flexibility by removing arterial wall tissue from the outer arterial layer. This leads to expansion of the narrowed vessel resulting in increased blood flow. We generated cuts in dissected arteries of sheep and pigs by photo-ablation with an ArF-Excimer Laser operating at a wavelength of 193 nm. During the cutting process the vessel diameter was monitored by measuring the running time of the laser induced pressure transients with a pressure transducer lying under the artery. A nearly linear increase of the diameter dependent on the residual wall thickness was found with a maximum increase of vessel diameter about 10%. We also observed that the arterial wall maintains stable to very small residual wall thicknesses i.e. deep cutting. To support the experiments and to test different geometries of tissue removal we performed FEM-Analysis. We simulated vessel deformation and the total strain depending on the depth, width and number of cuts in the outer artieral wall. We also found a significant increase of the "lumen" in a model with atherosclerotic shape obtained from a histological section.
To improve the efficiency of laser phacoemulsification we developed a compact, high-repetition-rate, high-average-power, diode-pumped, 2.94 μm TIR-cavity Er:YAG laser system. Laser parameters of 19.4% slope efficiency, 5 W of average output power at up to 300 Hz repetition rate and more than 1.5 W at 1 kHz are demonstrated. The special design results in low thermal lenses of 1.9 Dpt/W. This is a good condition for high laser system scalability.
Due to the low energy threshold of photodisruption with fs laser pulses, thermal and mechanical side effects are limited to the sub μm range. The neglection of side effects enables the use of ultrashort laser pulses in a broad field of medical applications. Moreover, the interaction process based on nonlinear absorption offers the opportunity to process transparent tissue three dimensionally inside the bulk.
We demonstrate the feasibility of surgical procedures in different fields of medical interest: In ophthalmology intrastromal cutting and preparing of corneal flaps for refractive surgery in living animals is presented. Besides, the very low mechanical side effects enables the use of fs-laser in otoralyngology to treat ocecular bones. Moreover, the precise cutting quality can be used in fields of cardiovascular surgery for the treatment of arteriosclerosis as well as in dentistry to remove caries from dental hard tissue.
Laser induced ultrasound provides a powerful tool for solving a major problem of laser cyclophotocoagulation, which is caused by difficulties in localization and determination of optical properties. Furthermore it adds the possibility of an online control mechanism for the process of coagulation of the ciliary body. We have developed a transducer system which is based on a fiber with 600 micrometers core diameter surrounded by a ring shaped piezoelectric PVDF detector. With this detector it is possible to localize the lateral position of the ciliary body on enucleated pigs and rabbit eyes as well as its depth. Our findings correspond well with histological sections of the measured area. Additionally, the changes in the tissue's optical properties induced by coagulation with a diode laser have been detected in real time.
In order to perform refractive surgery, fs-laser pulses of 130-fs pulse duration were focused into animal cornea. By focusing the radiation down to spot-sizes of 5 micrometers , very precise cuts could be achieved inside the treated eyes, accompanied with minimum collateral damage to the tissue by thermal or mechanical effects. Due to these low side effects, micrometers -cutting precisions at preparing of corneal flaps and lenticules can be achieved. Thus, animal studies with 8 living rabbits were performed, in which intrastromal lenticules and flaps were created and extracted. Wound healing reactions were studied by histo- pathological analysis of the treated eyes in intervals at one, 7, 14 and 28 days after surgery. The treated eyes showed mild wound healing reactions with comparable results to what is known from Excimer-LASIK. Moreover the use of the fs-laser offered new possibilities in preparation of corneal flaps, providing advantages to the use of the mechanical keratome.
The optical properties of human skin in the UV-range are not exactly known. Furthermore, the precise wavelength dependency of important photobiological processes (such as induction of skin cancer) could not be settled yet, either. A better knowledge of the optical properties is necessary in order to achieve a better understanding of UV-induced effects on human skin. Optoacoustics is a new approach to investigate the wavelength dependent optical properties of human skin in the UV-range. This technique allows non-invasive measurements on human skin in vivo, that are indispensable to gain meaningful results concerning the processes induced by UV-radiation in the living tissue. First attempts at measuring UV-induced optoacoustic transients of human skin in vivo and tissue phantoms with a new detector are shown. For analysis, fitting of simulated data onto the experimental data is applied in order to improve the determination of optical properties. First measurements of wavelength dependent optical properties in the UVB-(280-315 nm) and UVA-II-range (290-330 nm) comparing stained artificial layers to human skin in vivo are presented.
For enhanced optoacoustic imaging in biomedical applications more than one-dimensional detection is required. Tomographic images comparable to ultrasound B-scans can be generated with linear arrays. This work presents a detection scheme for such an array based on piezoelectric films. Each single detector consists of a ring shaped active area with a diameter of less than one millimeter, which leads to a high lateral resolution. Because of the small dimensions of the systems it is suitable for applications with limited accessibility like ophthalmic or endoscopic use. The sensitivity of a single detector is close to 0.5 mV/bar. First measurements on layered tissue phantoms made of gelatine and absorbing films show the potential of such an array for depth profiling as well as for two-dimensional imaging of simple structures.
Ultrafast lasers have become a promising tool for micromachining and extremely precise ablation of all kinds of materials. Due to the low energy threshold, thermal and mechanical side effects are limited to the bu micrometers range. The neglection of side effects enables the use of ultrashort laser pulses in a broad field of medical applications. Moreover, the interaction process based on nonlinear absorption offers the opportunity to process transparent tissue three dimensionally inside the bulk. We demonstrate the feasibility of surgical procedures in different fields of medical interest: in ophthalmology intrastromal cutting and preparing of cornael flaps for refractive surgery in living animals is presented. Besides, the very low mechanical side effects enables the use of fs- laser in otoralyngology to treat ocecular bones. Moreover, the precise cutting quality can be used in fields of cardiovascular surgery for the treatment of arteriosklerosis as well as in dentistry to remove caries from dental hard tissue.
In order to perform refractive surgery, fs-laser pulses were focused into animal cornea. During histo-pathological analysis by light- and transmission-electron-microscopy new side effects of fs-photodisruption were found. Due to the high intensities at the focal region, self focusing followed by further nonlinear effects is observed, leading to streak formation inside the cornea and several micro-breakdowns on the sub-micron scale. The nature and origin of these streaks are further investigated.
The feasibility of optoacoustic imaging was investigated for ophthalmologic application in the treatment of glaucoma. Difficulties in the treatment with laser cyclophotocoagulation are mainly due to uncertainties in the localization of the ciliary body. With laser optoacoustics it is possible to localize the position of the ciliary body on enucleated porcine and rabbit eyes. Additionally, the changes in the optical properties of the tissue induced by coagulation with a diode laser were observed.
In this work, we show the capability of laser optoacoustics to localize the position of the ciliary body on enucleated porcine and rabbit eyes. Our findings correspond well with histological sections of the measured area. Different wavelengths for an optoacoustical detection system in combination with laser cyclophotocoagulation have been compared taking grayscale images of the region of interest of rabbit and porcine eyes for various wavelengths in the NIR spectral range. Additionally, the changes in the optical properties of the tissue induced by coagulation with a diode laser were observed. First online measurements of the changes due to coagulation show that the method of laser optoacoustics is suitable for an online therapy control system.
Focussing femtosecond laser pulses into a transparent media, such as corneal tissue, leads to optical breakdown, generation of a micro-plasma and, thus, a cutting effect inside the tissue. To proof the potential of fs-lasers in refractive surgery, three-dimensional cutting within the corneal stroma was evaluated. With the use of ultrashort laser pulses within the LASIK procedure (laser in situ keratomileusis) possible complications in handling of a mechanical knife, the microkeratome, can be reduced by using the treatment laser as the keratome itself. To study woundhealing effects, animal studies were carried out in rabbit specimen. The surgical outcome was analyzed by means of histological sections, as well as light and scanning electron microscopy. Dependencies on the dispersion caused by focussing optics were evaluated and optimized. Thus, pulse energies well below 1 (mu) J were sufficient to perform the intrastromal cuts. The laser pulses with a duration of 180 fs and energies of 0.5-100 (mu) J were provided by a modelocked frequency doubled erbium fiber-laser with subsequent chirped pulse amplification in a titanium sapphire amplifier at up to 3 kHz.
In this work the dependence of the threshold energy and pulse duration for photodisruption was studied between 100 and 1500 femtoseconds using an erbium fiber laser system with subsequent chirped pulse amplification in a titanium-sapphire system. Cutting effects were analyzed in animal specimens like rabbit cornea and porcine eye globes. To perform intrastromal cuts, the laser beam was guided through a computer controlled delivery system. The treated tissue was analyzed by histological sections and by means of light and electron microscopy. Pressure transients were measured at the endothelium during the cutting procedure. Bubble formation and dynamics were studied using flash photography. The resulting gas bubbles were analyzed to their content, and photodissociation could be noticed. As a result of these studies a set of optimized parameters for pulse duration, scanning algorithm, pulse energy and the focusing and beam delivery system could be found. The thermal damage zone in the irradiated tissue was found to be smaller than 1 micrometer and maximum pressure transients at the endothelium could be determined to be in the range of some hundred millibars. Moreover, performing of intrastromal cuts and creation of a lenticule for achieving a refractive effect in the eye was demonstrated. The investigations show, that fs-pulses are a very precise tool for refractive surgery.
In this work, the localization of the ciliary body region of porcine eyes with laser optoacoustics was realized. The principle of laser optoacoustics combines the advantages of ultrasound and optical tomography. The absorption of short laser pulses in the near infrared generates thermoelastic stress waves in the ciliary muscle and the underlying pigmented epithelial layers. These stress transients were detected at the surface of the sclera with piezoelectric transducers in order to analyze the structure. The optoacoustical measurements of the ciliary body region at different positions from the corneoscleral limbus were composed to a grayscale image, which could be qualitatively compared to histological sections of this region. In these images the pigmented layers can be localized. The influence of the detectional wavelength on the optoacoustical signal in the NIR between 750 and 1052 nm was investigated. Furthermore, changes in the optoacoustic signal due to coagulation with a diode laser using typical therapeutical parameters could be observed.
In this work, the feasibility of optoacoustic tomography to act as diagnostic tool in laser cyclophotocoagulation was investigated. The experiments were performed with enucleated porcine and rabbit-eyes. The absorption of short laser pulses in the wavelength range between 750 and 1000 nm generated thermoelastic stress waves in the ciliary muscle and the underlying pigmented epithelial layers. These stress transients were detected at the surface of the sclera with piezoelectric transducers in order to analyze the structure. Depths of the ciliary body could be determined at different distances from the corneoscleral limbus. Furthermore, changes in the optoacoustic signal due to coagulation with a diode laser using typical therapeutical parameters could be observed.
The Feasibility of refractive surgery with ultrashort laser pulses in the fs-regime was studied. The Experiments were performed with an Erbium fiberlaser oscillator, which was amplified by means of chirped pulse amplification in a Titanium-Sapphire regenerative amplifier. With this system pulse durations of 180 fs and pulse energies of up to some hundred (mu) J were possible. The photodisruption was performed in corneal tissue in order to create corneal incisions and lamellars. Surface quality and thermal damages of the cuts inside the tissue were studied with the help of micromorphological analysis, such as scanning electron microscopy (SEM) and histological sections. High level precision along with minimal thermal damage could be observed. Saline solution as a model substance was used for studying side effects as bubble formation and pressure transients. The pressure measurements were performed using broadband acoustic transducers. Gas bubbles were analyzed with respect to formation, dynamics, interaction and composition of different gas constituents with the help of flash photography and gas chromatography. The results demonstrate the minimal invasive treatment of corneal stroma by fs-photodisruption. Studies of secondary effects indicate, that additional processes have to be considered.
Laser radiation in the mid infrared region is a promising tool for soft tissue processing due to its strong absorption in water. In this study, a cw Tm:YAG laser emitting at 2.06 micrometer was used for basic investigations. The laser has a maximum output power of more than 30 W. Fresh pig skin, liver and heart tissue were irradiated at different intensities (up to 65 kW/cm2) and irradiation times (400 ms to 20 s). As a delivery system a 400 micrometer quartz fiber was used in contact as well as in non-contact mode. The irradiated samples were examined by light and electron microscopy. The ablation efficiency on soft tissue was determined to about 0.3 mg/J. The thermal induced damage covered a region of 100 to 500 micrometers lateral and 500 to 1000 micrometers axial to the crater. With regard to its compact dimensions and high efficiency the cw Tm:YAG laser appears to be a promising alternative to the CO2 laser especially when fiber transmission of the radiation is required.
Pulsed photothermal radiometry (PPTR) is a contact free nondestructive method of material inspection. It also is a promising tool for in vivo tissue spectroscopy and on-line dosimetry. For this approach, one has to consider light scattering and thus a complicated signal interpretation. Different models of light propagation in scattering media are compared, such as Monte Carlo simulation (MCS) and various approaches of diffusion theory. Simulated PPTR signals with these calculations are compared. The validity of the different models in order to describe PPTR measurements correctly is discussed.
These studies are directed towards an on-line monitoring of tissue optical properties during medical laser diagnostics and therapy. The method of detection is based upon photoacoustic measurements. The detection of the laser induced acoustic transients was realized by means of piezoelectric transducers. Results concerning the optical properties of layered tissue- like samples are presented for two experimental configurations, i.e. transmission mode and reflection mode. The corresponding transients are calculated analytically and account for different influences which contribute to the signal, such as the laser parameters. Especially the effect of optoacoustic diffraction was investigated experimentally and verified theoretically. Its influence could be eliminated by applying fitting functions to the detected transients which take into account the distorting effect of diffraction.
Photoablative tissue processing at the wavelength (lambda) approximately equals 3 micrometer is of great interest in many medical applications but not yet really understood. A mathematical model of the photoablation process using a free- running infrared laser has been developed. It includes evaporation and thermoelastic pressure generation and was solved using the finite-element-method. Simulated thermoelastic pressure transients are in good agreement with the experiment. It has been shown, that the temperature dependence of the absorption and the volumetric expansion cannot be neglected. With higher laser intensities strong recoil pressure transients (greater than or equal to 100 bar) and strong thermoelastic pressure transients due to a partially evaporation are given. For this reason a new model including large tissue expansions, tissue overheating and recoil induced pressure transients has been developed and presented.
In this study the ablation characteristics and the wound healing process of rabbit cornea irradiated with a Q- switched Er:YAG laser was evaluated. The laser, emitting at 2.94 micrometers wavelength, has a pulse width of 100 ns. The spot size on the corneal surface was 1 mm in diameter at a fluence of 750 mJ/cm2. The laser beam was applied by a `flying spot' mode, performing refractive ablations of -7 to -8 dpt. As a biological model, the corneas of 9 rabbits were irradiated. The post-treatment follow-up was as long as 39 days. The treated corneas were investigated by light and electron microscopy. The wound healing on rabbit cornea of the Q-switched Er:YAG laser radiation in corneal tissue processing resembles to what is known from ArF- excimer laser application. To shorten the pulse width by means of Q-switching is one major key to the successful application of the Er:YAG laser for PRK.
KEYWORDS: Tissue optics, Photoacoustic spectroscopy, Signal detection, Acoustics, Pulsed laser operation, Transducers, Signal processing, Signal attenuation, Near field optics, Optical properties
Tissue optical properties are determined by an interpretation of pressure transients generated by irradiation with pulsed laser light. These pressure signals are detected using piezoelectric transducers. The signals are influenced by numerous processes during generation, propagation and detection. To study each process in particular, specific experiments were performed using biological tissue in vitro as well as different substances with various absorption and scattering properties. Especially acoustical diffraction effects cause a strong distortion of the shape and spatial oscillations in the amplitude of the transients. These phenomenons were investigated experimentally and verified theoretically.
In the mid IR region, where water acts as the main chromophore in soft tissue, pulsed erbium lasers emit radiation with sufficient energy for photoablative tissue processing. In this study, the ablation characteristics of these lasers were analyzed in order to evaluate their potential for clinical application in corneal and intraocular surgery. Histopathological examination of thermal side-effects was performed on porcine corneas. Ablation rates were investigated by on-line depth profiling in free-running and in q-switched mode. Structure and intensity of laser induced acoustic transients were analyzed by piezo-electric PVDF-films.
The knowledge of laser tissue interaction, the light propagation in tissue and thus its optical properties are of fundamental importance for both laser treatment and diagnostics. We use the method of pulsed photothermal radiometry (PPTR) to evaluate optical or thermal properties of tissue-like phantoms. Because its a non-destructive and non-contact technique its a promising tool for tissue optics in vivo. As phantoms we used gelatine of known optical properties with ink added as absorber. Once the thermal features are known, one can determine the optical properties of the sample and vice versa. PPTR investigations of absorption coefficients were compared to other methods such as time resolved stress detection and optical transmission measurements. PPTR is a suitable tool to perform spectroscopy under conditions of high pressure and/or temperature and can therefore be used to investigate the ablation process.
The ablation of soft tissue with free-running infrared lasers of low intensity is a thermal evaporation process. Nevertheless, the influence of the temperature dependent optical absorption is not yet understood. A detailed model of the photoablation process has been developed which was solved by the finite element method. Ablation rates and thermal damages are calculated and compared with experimental values. It has been shown, that the influence of the temperature dependent absorption coefficient cannot be neglected.
Correction of myopia with the ArF-excimer laser (PRK) sometimes leads to a so called 'central island' formation on the anterior corneal surface. The attenuation of the laser beam by the ablation plume might be one reason for this phenomenon. The attenuation properties of the ablation plume were investigated by a probe beam parallel to the surface of the tissue probe. By varying the laser parameters (fluence, repetition rate, spot size) and the target tissue (cornea, PMMA) the attenuation of the probe beam was measured time and spatial resolved. As a result of this study, a significant influence of the removal process due to scattering and absorption within the ablation plume can be assumed as a function of repetition rate, spot size and air flow on the tissue surface.
For an interpretation of pressure signals detected by piezoelectric PVDF-foils several parameters are of importance. These are for example pressure pulse duration and rise times, active area of the film and specifications concerning the high-frequency technique of the experimental setup. Using an input-resistance of 1 M(Omega) at the storage-oscilloscope leads to reflections of the signal which superimpose each other when reflected a second time at the foil. This might cause an apparent increase of the amplitude. The utilization of a longer cable allows a separation of each reflected pulse, but it causes the problem, that the cable's input- impedance approaches its characteristic impedance of 50 (Omega) . This will lead to a time derived signal as well as using 50 (Omega) as input-resistance at the scope. Therefore a direct measurement of short pressure transients requires a very short cable and an input-resistance of 1 M(Omega) . Otherwise the measured signal allows no proportional relation between pressure transient and measured voltage signal. In the latter case a frequency-dependent correction of the signal becomes necessary. This has been developed in this paper by means of Fast Fourier Transform algorithm. After correction in the frequency-domain the signal is transformed back into the time-domain.
Laser ablation of corneal tissue by mid-infrared laser radiation was studied in dependence of the laser wavelength and the pulse duration. The thermally and mechanically laser induced damage was determined by light microscopy (LM) and scanning electron microscopy (SEM). Fresh porcine eyes were irradiated with three different lasers: Er:YAG ((lambda) equals 2.94 micrometers ), Er:YSGG ((lambda) equals 2.79 micrometers ) and Ho:YAG ((lambda) equals 2.1 micrometers ). The experiments were performed with both, free running (pulse duration (tau) equals 200 microsecond(s) ) and Q-switched pulsed ((tau) equals 100 ns) lasers. The extent of thermally damaged tissue was found to be the same for Er:YAG and Er:YSGG lasers: 8 - 45 micrometers and 2 - 10 micrometers , with the long and the Q-switched pulses, respectively. The Ho:YAG laser induced coagulation zones were 200 - 600 micrometers thick in the free running and 40 - 80 micrometers in the Q-switched mode. The ablation efficiency of Er-lasers was between 4 and 13 times higher than that of the Ho:YAG.
Frontiers in Ultrafast Optics: Biomedical, Scientific, and Industrial Applications XI
23 January 2011 | San Francisco, California, United States
Frontiers in Ultrafast Optics: Biomedical, Scientific and Industrial Applications X (Formerly: Commercial and Biomedical Applications of Ultrafast Lasers)
24 January 2010 | San Francisco, California, United States
Advanced Technology and Instrumentation in Otolaryngology: Lasers, Optics, Radio Frequency, and Related Technology
23 January 2010 | San Francisco, California, United States
Commercial and Biomedical Applications of Ultrafast Lasers IX
25 January 2009 | San Jose, California, United States
Advanced Technology and Instrumentation in Otolaryngology: Lasers, Optics, Radio Frequency, and Related Technology
24 January 2009 | San Jose, California, United States
Advanced Technology and Instrumentation in Otolaryngology: Lasers, Optics, Radio Frequency, and Related Technology
19 January 2008 | San Jose, California, United States
Therapeutic Laser Applications and Laser-Tissue Interactions
18 June 2007 | Munich, Germany
Therapeutic Laser Applications and Laser-Tissue Interactions II
12 June 2005 | Munich, Germany
Femtosecond Laser Applications in Biology
29 April 2004 | Strasbourg, France
Therapeutic Laser Applications and Laser-Tissue Interactions
Access to the requested content is limited to institutions that have purchased or subscribe to SPIE eBooks.
You are receiving this notice because your organization may not have SPIE eBooks access.*
*Shibboleth/Open Athens users─please
sign in
to access your institution's subscriptions.
To obtain this item, you may purchase the complete book in print or electronic format on
SPIE.org.
INSTITUTIONAL Select your institution to access the SPIE Digital Library.
PERSONAL Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.