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At least 55 records · Page 3Linked to original sources

Brain NADH redox state monitored in vivo by fiber optic surface fluorometry.

A new approach for the evaluation of brain energy metabolism in awake animals became possible as UV transmitting optical fibers became available. A variety of surface fiber optic fluorometers / reflectometers which were developed during the past decade enabled the monitoring of intramitochondrial NADH redox state in unanesthetized animals. The bundle of flexible fibers was connected to the brain via a cemented light guide holder implanted epidurally. The two signals obtained, 366 nm reflectance and 450 nm fluorescence, are subjected to various artifacts not connected to the intramitochondrial NADH redox state. In our system, the effects of movement artifacts and changes in blood oxygenation are negligible while the effects of tissue absorption or blood volume changes are considerable and could be minimized by subtraction of the two signals (1:1 ratio) providing the corrected fluorescence signal. The brain was exposed to various physiological and pathological conditions which resulted in the increase or decrease in the level of NADH. Under anoxia, hypoxia and ischemia, oxygen availability decreased and the metabolic state of the brain became more reduced (state 4-5 transition). When the brain was activated by seizures, spreading depression of hyperbaric oxygenation NADH became more oxidized (state 4-3 transition).

Animals↗

Performance evaluation of cylindrical fiber optic light diffusers for biomedical applications.

BACKGROUND AND OBJECTIVES: Dosimetry for intracavity and interstitial light delivery requires next to the knowledge of tissue optical properties and models describing light propagation in tissue also exact knowledge of the spatial light source emission characteristics. However, the emission characteristics of cylindrical diffusers are often ill defined by the manufacturer, and not regularly determined by the end user, thus limiting the attainable dosimetry accuracy. STUDY DESIGN/MATERIALS AND METHODS: Commercial cylindrical diffusers, with active diffusive lengths of 1-2 cm and outer diameters of </=1 mm, were evaluated regarding their photometric emission characteristics. In addition to traditional tests of homogeneity such as light intensity emitted along the diffuser length and around its circumference (polar angle), azimuth emission was also measured. RESULTS: It was demonstrated that the light emission distribution is specific to the diffuser manufacturer, and while good polar isotropy is often attained, azimuth isotropy can vary by more than a factor of 5. In hollow organs the latter can result in over treatment of tissue distal to the actual placement of the cylindrical diffusers. CONCLUSIONS: To enable light dosimetry and treatment planning for applications such as photodynamic therapy or interstitial laser hyperthermia or photocoagulation, standardization of emission properties reporting for optical fiber diffusers is required.

Equipment Design↗

Fiber optic biosensor using Chlorella vulgaris for determination of toxic compounds.

A new biosensor is constructed for the detection of some herbicides based on kinetic measurements of chlorophyll-a fluorescence in Chlorella vulgaris cells. The microalgae are immobilized on removable membranes placed in front of the tip of an optical fiber bundle inside a homemade microcell. C. vulgaris was easily cultivated in laboratory and very sensitive to herbicides that effect the photosynthesis process. The response of the algal biosensor is studied in terms of detection limits, reversibility, and long-term activity. The effects of temperature and pH are also reported. The biosensor can be used to measure the concentration of a toxic chemical in the form of a single drop or dissolved in a continuous flow. The detection of 0.1 microg small middle dotL(-1) of a single herbicide as is required by European Community legislation for drinking water is possible with this algal biosensor especially for atrazine, simazine, and diuron.

Biosensing Techniques↗

CT-guided percutaneous laser disc decompression with Ceralas D, a diode laser with 980-nm wavelength and 200-microm fiber optics.

The aim of this study was to evaluate the compact, portable Ceralas-D diode laser (CeramOptec; 980 + 30 nm wavelength, 200-microm optical fiber) concerning clinical usefulness, handling, and clinical results in the CT-guided treatment of herniated lumbar discs. The positioning of the canula in intradiscal space, the placement of the laser fiber into the disc through the lying canula, and the vaporization itself were carried out under CT-guidance. Due to the thin fiber optic, it was possible to use a thin 23-gauge canula. The laser procedure was performed in 0.1- to 1-s shots with 1-s pulse pause and 4-W power output. A total of 1650-2300 J was applied on each percutaneous laser disc decompression (PLDD). Results in 26 patients were established with a visual-analogue scale (VAS). On the follow-up examinations, 46% of the patients were absolutely pain free ( > 85 % VAS) and fully active in everyday life after 4 postoperative weeks. Thirty-one percent of patients were relieved of the leg pain but had occasional back pain without sensorimotor impairment. Fifteen percent sensed a slight alleviation ( > 50% VAS) of the radiate pain. Eight percent did not experience radicular or pseudo-radicular pain alleviation (< 25% VAS). Cerales-D proves to be an efficient tool for CT-guided PLDD on non-sequestered herniated lumbar discs.

Adult↗

Synthesis and fluorescence studies of thiazole orange tethered onto oligonucleotide: development of a self-contained DNA biosensor on a fiber optic surface.

Thiazole orange dyes were derivatized with ethylene glycol linkers of various lengths, and were covalently linked to the 5' end of the oligonucleotides after solid-phase synthesis. The labeled oligonucleotides exhibited enhanced fluorescence upon hybridization to complementary DNA sequences at the surfaces of optical fibers, providing for a self-contained labeling strategy. It was determined that the melt temperatures of DNA hybrids using one mixed polypyrimidine base oligonucleotide sequence were dependent on the length of the tethers, and that the melt temperature could be shifted by more than 10 degrees C when tethers were introduced.

Base Sequence↗

Measurement of blast wave by a miniature fiber optic pressure transducer in the rat brain.

Exposure to blast wave that is generated during an explosion may result in brain damage and related neurological impairments. The aim of this study was to investigate pressure changes induced by exposure to blast inside the rat brain. For intracranial pressure measurement we used a miniature optic fiber sensor (o.d. 550 microm) with a computer recording system. The sensor was placed in the third cerebral ventricle of anesthetized rats exposed to 40 kPa blast wave in a pneumatic-pressure driven shock tube. Short pressure waves lasting several ms were detected inside the brain with the magnitude that might result in nervous tissue damage.

Animals↗

Fiber-optic microsensor array based on fluorescent bulk optode microspheres for the trace analysis of silver ions.

An optical microsensor array is described for the rapid analysis of silver ions at low parts per trillion levels. Because the ionophore o-xylylenebis(N,N-diisobutyldithiocarbamate) (Cu-I) was reevaluated and shown to exhibit excellent selectivity for silver ions, ion-selective electrode (ISE) membranes were optimized and found to exhibit the lowest reported detection limit so far (3 x 10(-10) M). A corresponding Ag+-selective fluorescent optical microsensor array for the rapid sensing of trace level Ag+ was then developed. It was fabricated using plasticized PVC-based micrometer-scale fluorescent microspheres that were produced via a sonic particle casting device. They contained 156 mmol/kg Cu-I, 10 mmol/kg 9-(diethylamino)-5-[4-(15-butyl-1,13-dioxo-2,14-dioxanodecyl) phenylimino]benzo[a]phenoxazine (chromoionophore VII, ETH 5418), 2.3 mmol/kg 1,1' '-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (internal reference dye), and 14 mmol/kg sodium tetrakis[3,5-bis(trifluoromethyl)phenyl]borate and were deposited onto the etched distal end of a 3200-microm-diameter optical fiber bundle. The microarray was characterized by fluorescence spectroscopy in samples containing 10(-12)-10(-8) M AgNO3 at pH 7.4, with selectivity characteristics comparable to the corresponding ISEs. The response time of the microsensor array was found to be less than 15 min for 10(-9) M AgNO3, which is drastically shorter than earlier data on optode films (8 h) and corresponding ISEs (30 min). A detection limit of 4 x 10(-11) M for Ag+ was observed, lower than any previously reported optode or silver-selective ISE. The microsensor array was applied for measurement of free silver levels in buffered pond water samples.

Calibration↗

Fiber-optic microsphere-based arrays for multiplexed biological warfare agent detection.

We report a multiplexed high-density DNA array capable of rapid, sensitive, and reliable identification of potential biological warfare agents. An optical fiber bundle containing 6000 individual 3.1-mum-diameter fibers was chemically etched to yield microwells and used as the substrate for the array. Eighteen different 50-mer single-stranded DNA probes were covalently attached to 3.1-mum microspheres. Probe sequences were designed for Bacillus anthracis, Yersinia pestis, Francisella tularensis, Brucella melitensis, Clostridium botulinum, Vaccinia virus, and one biological warfare agent (BWA) simulant, Bacillus thuringiensis kurstaki. The microspheres were distributed into the microwells to form a randomized multiplexed high-density DNA array. A detection limit of 10 fM in a 50-microL sample volume was achieved within 30 min of hybridization for B. anthracis, Y. pestis, Vaccinia virus, and B. thuringiensis kurstaki. We used both specific responses of probes upon hybridization to complementary targets as well as response patterns of the multiplexed array to identify BWAs with high accuracy. We demonstrated the application of this multiplexed high-density DNA array for parallel identification of target BWAs in spiked sewage samples after PCR amplification. The array's miniaturized feature size, fabrication flexibility, reusability, and high reproducibility may enable this array platform to be integrated into a highly sensitive, specific, and reliable portable instrument for in situ BWA detection.

Biological Warfare↗

Nitric oxide-releasing xerogel-based fiber-optic pH sensors.

A xerogel-based optical pH sensor capable of releasing low levels of nitric oxide (NO) and measuring changes in solution pH is reported. Through simple dip-coating procedures, aminoalkoxysilane-based xerogel films modified with N-diazeniumdiolate NO donor precursors and the fluorescent pH indicator seminaphthorhodamine-1 carboxylate (SNARF-1) were sequentially deposited onto optical fibers. The resulting sensors were characterized by fast and linear response to pH throughout the physiological range (pH 7.0-7.8). Real-time chemiluminescence measurements confirmed that the presence of the overlying SNARF-1-containing TMOS layer did not have an inhibitory effect on N-diazeniumdiolate formation or NO release, and the NO-releasing coatings were capable of maintaining NO fluxes >0.4 pmol/cm(2) s up to 16 h. In vitro blood compatibility studies using porcine platelets confirmed the expected thromboresistivity of the NO-releasing xerogel coatings.

Animals↗

Fluorescent fiber-optic calcium sensor for physiological measurements.

A new optical sensor based on covalent immobilization of a newly synthesized calcium-selective, long-wavelength, fluorescent indicator has been constructed, with a response dynamic range optimal for physiological measurements. Immobilization occurs via photoinitiated copolymerization of the indicator with acrylamide on the distal end of a silanized 125 micrograms diameter multimode optical fiber. The working lifetime of this sensor is limited only by photobleaching of the indicator. Due to the inherent hydrophilic nature of the acrylamide polymer, the response time of this new sensor is governed by simple aqueous diffusion of the ionic calcium. This results in sensor response times fast enough to monitor some concentration fluctuations at physiological rates. The ability to monitor calcium concentration fluctuations in a high background level of magnesium is also demonstrated with a calculated selectivity of 10(-4.5).

Biosensing Techniques↗

High-performance fiber-optic pH microsensors for practical physiological measurements using a dual-emission sensitive dye.

A fast and durable ratiometric pH microoptode that is highly accurate, precise, sensitive, reversible, and reproducible over the physiological ranges of pH, ionic strength, and temperature has been developed. The sensing site consists of 5 (and 6)-carboxynaphthofluorescein (CNF) entrapped in a polyacrylamide gel matrix via photopolymerization at the silanized end of an optical fiber with a diameter of 2 (pulled) or 125 microns (unpulled). The optode's precision for the pH 6.3-8.4 range in rat embryos, sera, or physiological (Earle's and Tyrode's) buffers was found to be better than +/- 0.03 pH unit. The pulled and unpulled optodes have respective upper limit response times of 1 and 400 ms for 1-pH-unit change. Over a 7-week period, they retain sensitivity for 600 and 10,000 measurements, respectively. Ratiometric measurements are made using a pH-sensitive emission peak on each side of an isosbestic point. The CNF microoptode is most suitable for biological applications because of its essentially linear response over the pH 7-8 range, its high sensitivity (slope about 2), and its almost perfect correlation with a pH macroelectrode. Furthermore, errors introduced by photobleaching, leaching, quenching, optode movement, and excitation source fluctuations are minimal.

Animals↗

Cellular applications of a sensitive and selective fiber-optic nitric oxide biosensor based on a dye-labeled heme domain of soluble guanylate cyclase.

Nitric oxide-selective sensors have been prepared with the heme domain of soluble guanylate cyclase (sGC), the only known receptor for signal transduction involving nitric oxide. Expressed in and purified from E. coli, the heme domain contains a stoichiometric amount of heme that has electronic and resonance Raman spectra almost identical to those of heterodimeric (native) sGC purified from bovine lung. The small size of the heme domain, its inability to bind oxygen, and its high affinity for nitric oxide make it well-suited for sensor applications. The heme domain has been labeled with a fluorescent reporter dye and changes in this dye's intensity are observed based on the sGC heme domain's characteristic binding of nitric oxide. The current sensors are prepared with 100-microns optical fiber but could also be prepared using submicrometer fiber tips. These sensors have fast, linear, and reversible responses to nitric oxide and are unaffected by numerous common interferents, such as oxygen, nitrite and nitrate. The sensor limit of detection is 1 microM nitric oxide. Glutathione has been shown to decrease the sensitivity of the sensor; however, the sensor response remains linear and can be calibrated on the basis of the glutathione concentration present in the biological environment of interest. The sensors have been used to measure extracellular nitric oxide production by BALB/c mouse macrophages. Minimal nitric oxide was produced by untreated cells, while high levels of nitric oxide were released from activated cells, e.g., 111 +/- 2 microM in a given cell culture.

Animals↗

Microfluidic devices fabricated in poly(methyl methacrylate) using hot-embossing with integrated sampling capillary and fiber optics for fluorescence detection.

High-aspect-ratio microstructures have been prepared using hot-embossing techniques in poly(methyl methacrylate) (PMMA) from Ni-based molding dies prepared using LIGA (Lithographie, Galvanoformung, Abformung). Due to the small amount of mask undercutting associated with X-ray lithography and the high energy X-ray beam used during photoresist patterning, deep structures with sharp and smooth sidewalls have been prepared. The Ni-electroforms produced devices with minimal replication errors using hot-embossing at a turn around time of approximately 5 min per device. In addition, several different polymers (with different glass transition temperatures) could be effectively molded with these Ni-electroforms and many devices (>300) molded with the same master without any noticeable degradation. The PMMA devices consisted of deep and narrow channels for insertion of a capillary for the automated electrokinetic loading of sample into the microfluidic device and also, a pair of optical fibers for shuttling laser light to the detection zone and collecting the resulting emission for fluorescence analysis. Electrophoretic separations of double-stranded DNA ladders Phi X174 digested with Hae III) were performed with fluorescence detection accomplished using near-IR excitation. It was found that the narrow width of the channels did not contribute significantly to electrophoretic zone broadening and the plate numbers generated in the extended length separation channel allowed sorting of the 271/281 base pair fragments associated with this sizing ladder when electrophoresed in methylcellulose entangled polymer solutions. The dual fiber detector produced sub-attomole detection limits with the entire detector, including laser source, electronics and photon transducer, situated in a single box measuring 3'' x 10" x 14".

Dinitrobenzenes↗

Simultaneous measurement of rat brain cortex PtO2 using EPR oximetry and a fluorescence fiber-optic sensor during normoxia and hyperoxia.

Electron paramagnetic resonance (EPR) oximetry is a promising, relatively non-invasive method of monitoring tissue partial pressure of oxygen (PtO(2)) that has proven useful in following changes in PtO(2) under various physiologic and pathophysiologic conditions. Optimal utilization of the method will be facilitated by systematic comparisons with other available methods. Here, we report on the absolute values and changes of rat brain PtO(2) using EPR oximetry and the OxyLite, an oxygen monitor based on fluorescence quenching, at adjacent locations in the same brain. EPR oximetry utilizes an implanted oxygen-sensitive material and reports tissue PtO(2) at the surface of the material. OxyLite measures PtO(2) using the fluorescence lifetime of a chromophore fixed to the tip of an optical fiber that is inserted into tissue. Measurements were made at a depth of 2-3 mm into the cortex during normoxia and during breathing of carbogen (95% O(2):5% CO(2)) followed by a return to normoxia. We conclude that in this study (1) PtO(2) values reported by the two methods are similar but not exactly the same, (2) both methods can record a baseline and rapid changes in PtO(2), (3) changes in PtO(2) induced by increasing FiO(2) from 0.26 to 0.95 (carbogen) were similar by the two methods and (4) in some rats breathing carbogen, absolute values of PtO(2) were above the sensitive range of the OxyLite method.

Animals↗

A fiber optic system for measuring dynamic mechanical properties of embryonic tissues.

Dynamic mechanical properties of embryonic tissues, including viscoelasticity and active motility, play a major role in morphogenesis. Mechanical abnormalities resulting from altered cytoskeletal chemistry presumably contribute to spina bifida and other birth defects, but research in this area has been limited by inadequate mechanical measurement techniques. This paper discusses aspects of embryo tissues that make them difficult to measure and describes a versatile new system developed and used to study the biomechanics of morphogenesis in amphibian embryos. It incorporates a flexible optical-fiber probe driven by piezoceramic elements to impose and measure feedback-controlled, two-dimensional, time-varying patterns of stress or strain in the tissue. Compression, tension, and shear geometries are possible. Capacitive displacement sensors provide the precision and low drift rates required for these measurements. Software control permits the system to perform both standard and customized rheological tests, including interactive ones in which test parameters change in response to measured tissue behavior. Sample size range is 50 microns to 5 mm. Displacement range is 60 microns with a resolution of 0.1 microns. Force range is 10 microN with a resolution of 0.02 microN. Frequency range is DC to 20 Hz.

Amphibians↗

A low-cost fiber-optic strain gage system for biological applications.

A new low-cost strain measurement system has been developed for the mechanical testing of biological soft tissues. The technique creates four spots of light on a tissue sample surface by piercing the tissue sample with two pairs of small light-conducting optical fibers (one pair for each axis of a biaxial stretch), terminated by high intensity infrared emitters. A large-area photodiode, located below the tissue sample, detects the light emitted from the two pairs of light-spots. Analog and digital circuitry analyze the current signal from the photodiode to determine the position of a light-spot in real time. Each infrared emitter is sequentially cycled "on" at a rate of 3 kHz and the resulting photodiode current signal, after being converted to a voltage signal, is held by an integrated circuit sample and hold amplifier. Analog differencing of pairs of light-spot voltage signals provides a final output proportional to the separation between coaxial light-spots.

Biomechanical Phenomena↗

Fiber-optic fluorescence correlation spectrometer.

Fluorescence correlation spectroscopy (FCS) is a sensitive technique used to probe size, concentration, flow velocity, and reaction kinetics in a dilute solution. Conventional FCS spectrometers achieve this sensitivity at the cost of using bulky optics. We demonstrate a technique that utilizes a single-mode optical fiber of 3.3 microm mode field diameter to perform FCS measurements. We demonstrate that the technique has adequate sensitivity to perform FCS measurements on fluorescent beads of 13 nm radius, and that the results agree with theoretical predictions. Our method potentially allows FCS to be extended to remote and in vivo applications.

Colloids↗

Fiber optic array biosensors.

Optical fiber arrays provide a powerful substrate for creating high-density sensing systems that can address a variety of biological problems. The fiber substrate can be used to create femtoliter wells that can be loaded with individual beads to create high-density arrays for multiplexed screening and analysis. In addition, living cells can be loaded into the arrays, and their individual responses monitored over long time periods, enabling functional screening of biologically active compounds. Adherent cells can be attached to the fiber substrate to provide a rapid method for observing cell migration and for screening anti-migratory compounds. Finally, individual enzyme molecules can be loaded into the array wells enabling single molecule detection via enzyme-catalyzed signal amplification.

Animals↗