First clinical application of a liquid-core light guide connected to an Er:YAG laser for oral treatment of leukoplakia.

For medical applications, erbium lasers are usually equipped with articulated mirror arms or special glass fibers. However, only with mirror arms is it so far possible to transmit high average powers or pulse energies in the region of 1 J to achieve suitable energy densities for fast tissue preparation. An alternative to the glass fiber systems mentioned above are liquid-core light guides. An extremely flexible liquid-core light guide was used to connect a dental Er:YAG laser system to an especially adapted dental laser applicator. The core liquid was continuously circulated during laser irradiation to transmit pulse energies up to 1.1 J. A modified laser handpiece was used for exemplary clinical treatment. The experimental setup with the highly flexible light guide was completed successfully, and its ease of handling for a dental surgeon was demonstrated in the clinical treatment of leukoplakia of the oral cheek mucosa. Complete ablation of the epithelium with the laser was performed. One year postoperatively, the patient remains disease-free. This article describes the technical realization of a liquid-core light guide system for medical applications. We report about the first successful clinical treatment of oral hyperkeratosis using this new light guide technology.

Mid-infrared laser spectroscopy for online analysis of exhaled CO.

Carbon monoxide (CO) detection in human breath is the focus of much research because of CO's possible use as a marker molecule for different diseases. Detecting CO in human breath remains a tough challenge because of the low concentrations of CO (ppm range) that must be detected. Another problem is that many other molecules, which can be found in human breath, can interfere in these measurements. Additionally, a time resolution of less than 1 s is needed to resolve the CO curve of an exhalation. In this study (13)CO instead of (12)CO concentration is measured. The measurements are performed with a cavity leak-out spectroscopy system. The system's properties match the above-mentioned specifications for measurements, even of the rare isotopologue with high specificity, a time resolution of less than 1 s and a detection limit of 7 ppb Hz(-1/2). Two investigations are presented here. The first is a measurement showing intraday changes between 9.5% and 23.3% of the exhaled CO level due to vigorous exercise. The second shows a long-term observation of the CO base level revealing natural variations in the recorded CO concentration. The covered time period is 25 weeks during which the differences between the minimum and maximum CO levels for each test case reached 84%.

Bone healing of the sheep tibia shaft after carbon dioxide laser osteotomy: histological results.

The aim of the study was to compare the histological results after complete osteotomies of the sheep tibia using either the prototype carbon dioxide (CO(2)) laser osteotome 'OsteoLAS' (n = 12) or an oscillating saw (n = 12). The laser parameters were as follows: wavelength 10.6 microm; energy of laser pulses 75-85 mJ; pulse duration 80 mus; pulse repetition rate 200 Hz; spot diameter 460 mum (1/e(2) level); radiant exposure 45-51 J/cm(2); peak irradiance 0.56-0.64 MW/cm(2). Both groups were divided into two subgroups (n = 6), and the animals were killed after 4 weeks or 12 weeks, respectively. Light and fluorescence microscopy with semiquantitative analysis and histomorphometry were performed to compare bone healing. Charring-free laser osteotomies were possible up to a depth of 20 mm with the short-pulsed CO(2) laser. The laser, however, required a significantly longer time to perform, and a wedge-shaped gap was present on the cis-cortex. After 4 weeks the osteotomy gaps were almost unchanged in both groups and filled with connective tissue. After 12 weeks the gaps were filled with newly formed bone in both groups. Primary gap healing was predominant in the laser group and longitudinal cortical remodelling in the control group. On a cellular level, no fundamental differences were observed for early and late stages of bone healing. Further research has to be focussed on improving the CO(2) laser ostetome in order to reduce the long duration of the laser osteotomy and the necessity of creating a wedge-shaped cut in thick bones.

Holographic topometry for a dense visualization of soft tissue for facial reconstruction.

Facial reconstruction can be used as a forensic technique to identify a person, when no other identification method is applicable. The facial soft tissue thickness applied to the skull is crucial when performing an accurate facial reconstruction. Historically, scientists developed several techniques to measure the soft tissue of the face. It was their aim, to build a database of a unique point-set, differentiated by gender, age, ethnic origin, BMI. All used a limited number of landmarks and an inaccurate measuring technique. We developed a contact-free and precise measuring technique, using low-dose CT and holographic data. Due to the extremely short exposure time, the holographic measurement is very precise. We lay out our first experiences to create a facial soft tissue layer map of the face.

High precision cochleostomy by use of a pulsed CO2 laser - an experimental approach.

A precise cochleostomy is a crucial step in cochlear implantation, particularly if residual hearing is to be preserved. A contactless ablation of the promontory bone by a pulsed CO(2) laser system seems to be a promising approach. The bone is removed by a scan head controlled laser beam in sequential scan cycles with a pulse rate of 50-100 mus. Digital picture analysis and pattern detection are used to identify the membranous lining of the cochlea. We achieved a bone ablation in a micrometer range per scan cycle with the laser. A perforation of the promontory bone could be detected by automatic pattern detection. The enhancement of automatic pattern detection can lead to a minimally invasive, function-preserving laser cochleostomy.

Improvements in shape-from-focus for holographic reconstructions with regard to focus operators, neighborhood-size, and height value interpolation.

This paper presents a shape-from-focus method, which is improved with regard to the mathematical operator used for contrast measurement, the selection of the neighborhood size, surface refinement through interpolation, and surface postprocessing. Three-dimensional models of living human faces are presented with such a high resolution that single hairs are visible.

Is exhaled carbon monoxide level associated with blood glucose level? A comparison of two breath analyzing methods.

The level of exhaled carbon monoxide (eCO) is considered a marker of oxidative stress in diabetes. Previous findings indicated that eCO levels correlated with blood glucose level. The aim of this work was to apply and compare two independent analyzing methods for eCO after oral glucose administration. Glycemia, eCO, and exhaled hydrogen were measured before and after oral administration of glucose. Six healthy nonsmoking volunteers participated. For eCO analysis, we used two methods: a commercially available electrochemical sensor, and a high-precision laser spectrometer developed in our laboratory. The precision of laser-spectroscopic eCO measurements was two orders of magnitude better than the precision of the electrochemical eCO measurement. eCO levels measured by laser spectrometry after glucose administration showed a decrease of 4.1%+/-1.5% compared to the baseline (p<0.05). Changes in the eCO measured by the electrochemical sensor were not significant (p=0.08). Exhaled hydrogen levels increased by 40% within the first 10 min after glucose administration (p<0.05). The previous finding that the glycemia increase after glucose administration was associated with a significant increase in eCO concentrations was not confirmed. We propose that previous eCO measurements with electrochemical sensors may have been affected by cross sensitivity to hydrogen.

NO detection in biological samples: differentiation of 14 NO and 15 NO using infrared laser spectroscopy.

Accurate characterization of the biochemical pathways of nitric oxide (NO) is essential for investigations in the field of NO research. To analyze the different reaction pathways of enzymatic and non-enzymatic NO formation, determination of the source of NO is crucial. Measuring NO-related products in biological samples distinguishing between (14)NO and (15)NO offers the opportunity to specifically analyze NO signaling in blood and tissue. The aim of this study was to establish a highly sensitive technique for the specific measurement of NO in an isotopologue-selective manner in biological samples. With the cavity leak-out spectroscopy setup (CALOS) a differentiation between (14)NO and (15)NO is feasible. We describe here the employment of this method for measurements in biological samples. Certified gas mixtures of (14)NO/N(2) and (15)NO/N(2) were used to calibrate the system. (14)NO2- and (15)NO2- of aqueous and biological samples were reduced in a triiodide solution, and the NO released was detected via CALOS. Gas-phase chemiluminescence detection (CLD) was used for evaluation. The correlation received for both methods for the detection of NO in the gas phase was r=0.999, p<0.0001. Results obtained using aqueous and biological samples verified that CALOS enables NO measurements with high accuracy (detection limit for (14)NO2- 0.3 pmol and (15)NO2- 0.5 pmol; correlation (14)NO: p<0.0001, r=0.975, (15)NO: p<0.0001, r=0.969). The CALOS assay represents an extension of NO measurements in biological samples, allowing specific investigations of enzymatic and non-enzymatic NO formation and metabolism in a variety of samples.

Intercomparison of infrared cavity leak-out spectroscopy and gas chromatography-flame ionization for trace analysis of ethane.

Comparison of two different methods for the measurement of ethane at the parts-per-billion (ppb) level is reported. We used cavity leak-out spectroscopy (CALOS) in the 3 microm wavelength region and gas chromatography-flame ionization detection (GC-FID) for the analysis of various gas samples containing ethane fractions in synthetic air. Intraday and interday reproducibilities were studied. Intercomparing the results of two series involving seven samples with ethane mixing ratios ranging from 0.5 to 100 ppb, we found a reasonable agreement between both methods. The scatter plot of GC-FID data versus CALOS data yields a linear regression slope of 1.07 +/- 0.03. Furthermore, some of the ethane mixtures were checked over the course of 1 year, which proved the long-term stability of the ethane mixing ratio. We conclude that CALOS shows equivalent ethane analysis precision compared to GC-FID, with the significant advantage of a much higher time resolution (<1 s) since there is no requirement for sample preconcentration. This opens new analytical possibilities, e.g., for real-time monitoring of ethane traces in exhaled human breath.

Computer-guided CO2-laser osteotomy of the sheep tibia: technical prerequisites and first results.

OBJECTIVE: The purpose of this study was to examine for the first time the feasibility of performing complete osteotomy of sheep tibia using a computer-guided CO2-laser osteotome, and to examine bone healing under functional loading. BACKGROUND DATA: Bone cutting without aggravating thermal side effects has been demonstrated with scanning CO2-laser osteotomy. Further research is necessary to develop a clinically usable laser osteotome, which may allow new types of bone surgical procedures. MATERIALS AND METHODS: The scanning parameters for performing tibial osteotomies were determined in preliminary ex vivo trials. Osteotomies were performed in the mid-diaphysis of sheep tibia using either the prototype laser osteotome (osteoLAS, study group; n = 12), or an oscillating saw (control group; n = 12). Both groups were divided into two subgroups each (n = 6), and the two groups were sacrificed after 4 and 12 wk. Radiographs were taken postoperatively and after 4, 8, and 12 wk to compare the course of bone healing. RESULTS: Laser osteotomies of sheep tibia up to a depth of 20 mm were possible without visible thermal damage to the bone. A sequential PC-controlled cut geometry with artificial widening of the osteotomy gap was required for a complete osteotomy. Both clinically and radiologically, the laser and control groups showed undisturbed primary gap healing. Bone healing was similar and undelayed after both laser osteotomy and osteotomy done by mechanical saw. CONCLUSIONS: Osteotomy of multi-layered bones with a scanning CO2-laser demonstrates clinical and radiological healing patterns comparable to those seen with osteotomy done by standard mechanical instruments. It is, however, a technically demanding procedure, and complete laser osteotomies of long bones are only reasonable in bones with a diameter <20 mm, which will likely restrict the use of this technique to bones 7-10 mm thick. Through the use of computer guidance, extremely precise osteotomies and sophisticated cut geometries are possible using this technique. For practical applications, precise control of the depth of laser cutting and easier manipulation of the osteotome are required.

Generation of digital textured surface models from hologram recordings.

Digital sensors and fast digital image processing facilitate the use of pulsed holography for 3D surface measurement of moving objects. The real image of a hologram is reconstructed optically. A sequence of high-resolution projection images of the real image with a varying distance to the hologram is recorded digitally. Focus detection in this image sequence by digital image processing yields the shape of the recorded object. The image intensity serves as a precise pixel-matching texture. An application of this concept is the generation of a textured 3D computer model of a facial surface from a portrait hologram.

Infrared laser spectroscopy for online recording of exhaled carbon monoxide-a progress report.

Despite the growing number of reports on breath CO measurements, the development of rapid and sensitive analysis techniques for measurements of this breath constituent still remains a challenge. We demonstrate the application of infrared laser spectroscopy for exhaled CO analysis. The breath samples are analyzed in real-time during single exhalations by means of cavity ring-down spectroscopy. This is an ultra-sensitive laser-based method for the analysis of trace gases with precision on the ppb level (parts per billion). The noise-equivalent CO level of this method is 7 ppb Hz(-1/2); the time resolution is around 1 s. The expirograms were recorded with exhalation flow rates varying from 4 l min(-1) up to 50 l min(-1). Alveolar phase (phase III) of expiration shows a remarkable flow-rate dependence. Also, expirograms were recorded after a breath holding time between 0 s and 60 s. The normalized slope of the alveolar plateau (S(n)) was determined, which is between 0.004 l(-1) and 0.15 l(-1).

Ultra sensitive trace gas detection for biomedical applications.

We present an overview of our recent progress on spectroscopic trace gas detection for biomedical applications. The latest developments of cavity-enhanced spectroscopy as well as magnetic rotation spectroscopy lead to unprecedented sensitivity and specificity. The current detection limits of our laser spectroscopic approaches are in the picomolar to nanomolar range, depending on the molecular compound. The time resolution of the measurements is down to the sub-second range. This very high sensitivity and time resolution open up exciting perspectives for novel analytical tasks in biomedical research and clinical diagnosis.

In vivo animal trials with a scanning CO2 laser osteotome.

BACKGROUND AND OBJECTIVES: We report first results of animal trials using an improved laser osteotomy technique. This technique allows effective bone cutting without the usual thermal tissue damage. STUDY DESIGN/MATERIALS AND METHODS: A comparative in vivo study on mandibles of seven canines was done with a mechanical saw and a CO(2) laser based osteotome with a pulse duration of 80 microseconds. The laser incisions were performed in a multipass mode using a PC-controlled galvanic beam scanner and an assisting water spray. RESULTS: A complete healing through a whole bony rearrangement of the osteotomy gap with newly build lamellar Haversian bone was observed 22 days after the laser operations under optimal irradiation conditions. CONCLUSIONS: An effective CO(2) laser osteotomy without aggravating thermal side effects and healing delay is possible using the described irradiation technique. It allows an arbitrary cut geometry and may result in new advantageous bone surgery procedures.

Iterative focus detection in hologram tomography.

Hologram tomography is a two-step method for three-dimensional topometry of extended objects. The first step consists of the hologram recording with a single laser pulse of 35 ns duration and storage in a photosensitive material. In the second step the hologram is optically reconstructed and digitized, which leads to a set of two-dimensional projections at different axial positions. A maximization of a focus measure has to be performed to extract the surface position out of the projections. Unlike with well-established methods, where the comparison of sharpness values is done parallel to the optical axis, we propose an iterative solution to perform the maximization along the direction of image formation, which is evaluated for each surface point individually. This leads to a better reproducibility of the surface in the off-axis regions.

Measurement of the coagulation dynamics of bovine liver using the modified microscopic Beer-Lambert law.

BACKGROUND AND OBJECTIVES: During heating, the optical properties of biological tissues change with the coagulation state. In this study, we propose a technique, which uses these changes to monitor the coagulation process during laser-induced interstitial thermotherapy (LITT). STUDY DESIGN/MATERIALS AND METHODS: Untreated and coagulated (water bath, temperatures between 35 degrees C and 90 degrees C for 20 minutes.) samples of bovine liver tissue were examined using a Nd:YAG (lambda = 1064 nm) frequency-domain reflectance spectrometer. We determined the time integrated intensities (I(DC)) and the phase shifts (Phi) of the photon density waves after migration through the tissue. From these measured quantities, the time of flight (TOF) of the photons and the absorption coefficients of the samples were derived using the modified microscopic Beer-Lambert law. RESULTS: The absorption coefficients of the liver samples decreased significantly with the temperature in the range between 50 degrees C and 70 degrees C. At the same time, the TOF of the investigated photos was found increased indicating an increased scattering. The coagulation dynamics could be well described using the Arrhenius formalism with the activation energy of 106 kJ/mol and the frequency factor of 1.59 x 10(13)/second. CONCLUSIONS: Frequency-domain reflectance spectroscopy in combination with the modified microscopic Beer-Lambert (MBL) is suitable to measure heat induced changes in the absorption and scattering properties of bovine liver in vitro. The technique may be used to monitor the coagulation dynamics during local thermo-coagulation in vivo.

Time resolved simultaneous detection of 14NO and 15NO via mid-infrared cavity leak-out spectroscopy.

We present a ring-down absorption spectrometer based on a continuous-wave CO laser in the mid-infrared spectral region near lambda = 5 microm. Using a linear ring-down cavity (length: 0.5 m) with high reflective mirrors (R = 99.988 %), we observed a noise-equivalent absorption coefficient of 3 x 10(-10) cm(-1)Hz(-1/2). This corresponds to a noise-equivalent concentration of 800 parts per trillion (ppt) for (14)NO and 40 ppt for (15)NO in 1 s averaging time. We achieve a time resolution of 1 s which allows time resolved simultaneous detection of the two N isotopes. The delta(15)N value was obtained with a precision of +/-1.2 per thousand in a sample with a NO fraction of 11 ppm. The simultaneous detection enables the use of (15)NO as a tracer molecule for endogenous biomedical processes.

Parts per trillion sensitivity for ethane in air with an optical parametric oscillator cavity leak-out spectrometer.

Spectroscopic detection of ethane in the 3-microm wavelength region was performed by means of a cw optical parametric oscillator and cavity leak-out. We achieved a minimum detectable absorption coefficient of 1.6 x 10(-10) cm 1/square root of Hz, corresponding to an ethane detection limit of 6 parts per trillion/square root of Hz. For 3-min integration time the detection limit was 0.5 parts per trillion. The levels are to our knowledge the best demonstrated so far. These frequency-tuning capabilities facilitated multigas analysis with simultaneous monitoring of ethane, methane, and water vapor in human breath.

The presence of nitrite during UVA irradiation protects from apoptosis.

Nitrite occurs ubiquitously in biological fluids such as blood and sweat, representing an oxidation product of nitric oxide. Nitrite has been associated with a variety of adverse effects such as mutagenicity, carcinogenesis, and toxicity. In contrast, here we demonstrate that the presence of nitrite, but not nitrate, during irradiation of endothelial cells in culture exerts a potent and concentration-dependent protection against UVA-induced apoptotic cell death. Protection is half-maximal at a concentration of 3 mM, and complete rescue is observed at 10 mM. Nitrite-mediated protection is mediated via inhibition of lipid peroxidation in a similar manner as seen with butylated hydroxytoluene, a known inhibitor of lipid peroxidation. Interestingly, nitrite-mediated protection is completely abolished by coincubation with the NO scavenger cPTIO. Using electron paramagnetic resonance (EPR) spectroscopy or Faraday modulation spectroscopy, we directly prove UVA-induced NO formation in solutions containing nitrite. In conclusion, evidence is presented that nitrite represents a protective agent against UVA-induced apoptosis due to photodecomposition of nitrite and subsequent formation of NO.

Small-volume frequency-domain oximetry: phantom experiments and first in vivo results.

We describe a new method to determine the oxygen saturation and the total hemoglobin content of tissue in vivo absolutely at small source-detector separations (<10 mm). Phase and mean intensity of modulated laser light of various wavelengths was measured at several predetermined source-detector separations in the frequency domain. From these measured quantities, the absorption coefficient was derived using the modified time-integrated microscopic Beer-Lambert law (MBL). In addition, the interaction volume of the photons was determined using a multi-layer Monte-Carlo model of human skin. To evaluate the method, we employed homogenous solid phantoms (consisting of TiO2 particles embedded in resin) with mean scattering and absorbing properties comparable to those of human skin. Furthermore, in vivo measurements were performed in a healthy volunteer to demonstrate that the technique is applicable for the determination of the oxygen saturation and the total hemoglobin content in the skin in vivo. The proposed technique is especially suited for the on-line determination of the oxygen saturation and total hemoglobin content in applications where small applicators are required (e.g., fetal oxygen monitoring sub partu).