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Optical Recordings (optical + recording)
Selected AbstractsDevelopment of an Optrode for Intramural Multisite Optical Recordings of Vm in the HeartJOURNAL OF CARDIOVASCULAR ELECTROPHYSIOLOGY, Issue 11 2003JONATHAN L. BYARS M.S. Introduction: Optical mapping of transmembrane potential (Vm) is an important tool in the investigation of impulse propagation in the heart. It provides valuable information about spatiotemporal changes of Vm that cannot be obtained by other techniques, but it presently is limited to measurements from the heart surfaces. Therefore, the goal of this work was to develop a technique for intramural multisite optical measurements of Vm using fiberoptic technology. Methods and Results: An optrode, a bundle of thin optical fibers, was developed for measuring intramural optical signals at multiple sites in the heart. The optrode consisted of seven fibers with diameter of 225 ,m arranged in a hexagonal pattern that were used to deliver excitation light to the myocardium, to collect the emitted fluorescence, and to project the light onto a 16 × 16 array of photodiode detectors. Rabbit hearts were stained with the Vm -sensitive dye RH-237. Fluorescence was excited using a 100-W Hg lamp. Intramural action potentials were recorded at multiple sites separated by 2 mm inside the left ventricle. Signal-to-noise (RMS) ratio was 21.2 ± 12 (n = 7) without averaging or ratiometry and with negligible cross-talk (<1.9%) between the neighboring photodiodes. The size of the recording area for an individual fiber was estimated at approximately 0.8 mm. Conclusion: These data demonstrate feasibility of multisite transmural measurements of Vm without signal averaging and ratiometry. This technique might become useful in studies of transmural impulse conduction during arrhythmias and defibrillation. (J Cardiovasc Electrophysiol, Vol. 14, pp. 1196-1202, November 2003) [source] Optical recordings of taste responses from fungiform papillae of mouse in situTHE JOURNAL OF PHYSIOLOGY, Issue 2 2001Yoshitaka Ohtubo 1Single taste buds in mouse fungiform papillae consist of ,50 elongated cells (TBCs), where fewer than three TBCs have synaptic contacts with taste nerves. We investigated whether the non-innervated TBCs were chemosensitive using a voltage-sensitive dye, tetramethylrhodamine methyl ester (TMRM), under in situ optical recording conditions. 2Prior to the optical recordings, we investigated the magnitude and polarity of receptor potentials under in situ whole-cell clamp conditions. In response to 10 mM HCl, several TBCs were depolarized by ,25 mV and elicited action potentials, while other TBCs were hyperpolarized by ,12 mV. The TBCs eliciting hyperpolarizing receptor potentials also generated action potentials on electrical stimulation. 3A mixture of 100 mM NaCl, 10 mM HCl and 500 mM sucrose depolarized six TBCs and hyperpolarized another three TBCs out of 13 identified TBCs in a taste bud viewed by optical section. In an optical section of another taste bud, 1 M NaCl depolarized five TBCs and hyperpolarized another two TBCs out of 11 identified TBCs. 4The number of chemosensitive TBCs was much larger than the number of innervated TBCs in a taste bud, indicating the existence of chemosensitivity in non-innervated TBCs. There was a tendency for TBCs eliciting the same polarity of receptor potential to occur together in taste buds. We discuss the role of non-innervated TBCs in taste information processing. [source] Effect of molecular architecture and size of mesogen on phase behavior and photoactive properties of photoactive liquid crystalline hyperbranched polyester epoxies containing benzylidene moietyJOURNAL OF POLYMER SCIENCE (IN TWO SECTIONS), Issue 2 2008V. Srinivasa Rao Abstract A series of photoactive liquid crystalline linear and hyperbranched polyester epoxies were synthesized by polyaddition of photoactive bis benzylidene alkanone diol monomers and terephthalic acid and trimesic acid respectively with good yield. The effect of molecular architecture (linear and hyperbranched), size of mesogenic unit (cyclic and acyclic units) on the physicochemical, thermal, mesogenic, and photoactive properties of hyperbranched polymers were studied and compared. Degree of branching of hyperbranched polymers was found to be in the range of 0.46,0.49. Monomers containing cyclic moieties only exhibited nematic mesophase, while all polymers exhibited typical nematic mesophase. Intermolecular photo cycloaddition reaction was studied by ultraviolet,visible spectra (UV,vis) and NMR spectroscopy and photo viscosity measurement of UV irradiated polymer solutions. Faster photo induced behavior of hyperbranched polymers containing acyclic alkanone moiety, as compared to polymers containing cycloalkanone moieties, was observed. The change in the refractive index was found to be in the range of 0.02,0.024. Substantial variation of refractive index indicates that this polymer could be used for optical recording. All the polymers were also found to be fluorescent in nature. © 2007 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 46: 552,563, 2008 [source] Development of glutamate receptors in auditory neurons from long-term organotypic cultures of the embryonic chick hindbrainEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 2 2009Carmen Diaz Abstract We used long-range organotypic cultures of auditory nuclei in the chick hindbrain to test the development of glutamate receptor activity in auditory neurons growing in a tissue environment that includes early deprivation of peripheral glutamatergic input, subsequent to removal of the otocyst. Cultures started at embryonic day (E)5, and lasted from 6 h to 15 days. Neuronal migration, clustering and axonal extension from the nucleus magnocellularis (NM) to the nucleus laminaris (NL) partially resembled events in vivo. However, the distinctive laminar organization of the NL was not observed. Glutamate receptor (GluR) activity was tested with optical recordings of intracellular Ca2+ in the NM. ,-Amino-3-hydroxy-5-methyl-4-isoxazoleproprionic acid (AMPA)/kainate receptors had Ca2+ responses with a time course similar to that in control slices. Peak amplitude, however, was significantly lower. N -methyl- d -aspartate (NMDA)-mediated Ca2+ responses were higher in 2-day cultures (E5 + 2d) than in E7 explant controls, returning later to control values. Metabotropic GluRs did not elicit Ca2+ responses at standard agonist doses. Blocking NMDA or AMPA/kainate receptors with specific antagonists for 10 days in culture did not limit neuronal survival. Blocking metabotropic GluRs resulted in complete neuronal loss. Thus, ionotropic GluRs are not required for NM neuronal survival. However, their activity during development is affected when neurons grow in an in vitro environment that includes prevention of arrival of peripheral glutamatergic input. [source] In vivo optical recordings of synaptic transmission and intracellular Ca2+ and Cl, in the superior colliculus of fetal ratsEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 6 2006Yoshiyuki Sakata Abstract Although the N -methyl- d -aspartate (NMDA) receptor is known to play a crucial role in activity-dependent remodeling of synaptic connections in the fetal superior colliculus (SC), its contribution to the electrical activity of fetal SC neurons has not been determined. Furthermore, whether ,-aminobutyric acid (GABA)-mediated inhibition occurs either as early as prenatal periods or only after eye opening has been controversial. We therefore performed optical recordings using voltage-, Ca2+ - and Cl, -sensitive fluorescent dyes to analyse synaptic transmission and changes in intracellular Ca2+ and Cl, in the SC of fetal rats that were still connected with the dams by the umbilical cord. Excitatory and inhibitory responses were evoked by focal SC stimulation. The excitatory synaptic responses are composed of early and late components. The early component was mediated by both non-NMDA and NMDA receptors, whereas the late component occurred mainly via NMDA receptors. Train pulse stimulation at higher currents was required for induction of the inhibition, which was antagonized by bicuculline, and blocking of the GABA-mediated inhibition by bicuculline uncovered masked excitatory synaptic responses. Focal SC stimulation induced increases in [Cl,]i and [Ca2+]i that were mediated by GABA-A receptors and mainly by NMDA receptors, respectively. GABA antagonists augmented SC-induced increases in [Ca2+]i. These results indicate that, in the fetal SC, excitatory and inhibitory synaptic transmissions occur before birth, that the NMDA receptor is a major contributor to excitatory synaptic transmission and increased [Ca2+]i, and that the GABA-A receptor is already functioning to inhibit excitatory neurotransmission. [source] Simultaneous Optical Mapping of Transmembrane Potential and Intracellular Calcium in Myocyte CulturesJOURNAL OF CARDIOVASCULAR ELECTROPHYSIOLOGY, Issue 5 2000VLADIMIR G. FAST Ph.D. Simultaneous Mapping of Vm and Cai2+. Introduction: Fast spatially resolved measurements of transmembrane potential (Vm) and intracellular calcium (Cai2+) are important for studying mechanisms of arrhythmias and defibrillation. The goals of this work were (1) to develop an optical technique for simultaneous multisite optical recordings of Vm and Cai2+, and (2) to determine the relationship between Vm and Cai2+ during normal impulse propagation in myocyte cultures. Methods and Results: Monolayers of neonatal rat myocytes were stained with fluorescent dye RH-237 (Vm) and Fluo-3AM (Cai2+). Both dyes were excited at the same wavelength range. The emitted fluorescent was optically separated into components corresponding to changes in Vm, and Cai2+ and measured using two 16 × 16 photodiode arrays at a spatial resolution of up to 27.5 ,m per diode and sampling rate of 2.5 kHz. The optical setup was adjusted so that there was no optical cross-talk between the two types of measurements, which was validated in experiments involving staining with either RH-237 or Fluo-3. The amplitude of Fluo-3 signals rapidly decreased during experiments due to dye leakage. Dye leakage was substantially reduced by application of 1 mM probenecid, a blocker of organic anion transport, which had no effect on action potential duration and only minor effect on conduction velocity. In double-stained preparations, during regular pacing Cai2+ transients had a rise time of 14.2 ± 2 msec, and they followed Vm upstrokes with a delay of 5.3 ± 1 msec (n = 9). Durations of Vm, and Cai2+ transients determined at 50% level of signal recovery were 54.6 ± 10 msec and 136 ± 8 msec, respectively. Application of 2 ,M nifedipine reduced the amplitude and duration of Cai2+ transients without significantly affecting conduction velocity. Conclusion: The results demonstrate feasibility of simultaneous optical recordings of Vm and Cai2+ transients with high spatial and temporal resolution. [source] Electrochemical detection of neurotransmitters in the gut wallNEUROGASTROENTEROLOGY & MOTILITY, Issue 11 2008P. Vanden Berghe Abstract, Cells interact with each other by releasing signalling molecules, which can activate or inactivate target cells. In order to understand how coordination results from this communication, accurate measurements of these signalling molecules are prerequisite. Several different techniques exist to monitor and quantify these compounds, including enzymatic and histochemical assays, electrophysiological and optical recordings. However, there has been little use of electrochemical recordings in gastroenterological research, although these are very fast and sensitive. Electrochemical techniques rely on the simple fact that electroactive molecules can be oxidized at a given potential. The currents, elicited by the oxidation, are directly proportional to the concentration of the compound. In the current issue of Neurogastroenterology and Motility, electrochemical detection was successfully applied to measure nitric oxide (NO) from intestinal preparations. Although there are some important specificity, timing and spatial aspects to consider, this direct NO-probing technique is definitely a great asset to the field of gastrointestinal research and advances our understanding of NO signalling in the intestinal wall. [source] Optical recordings of taste responses from fungiform papillae of mouse in situTHE JOURNAL OF PHYSIOLOGY, Issue 2 2001Yoshitaka Ohtubo 1Single taste buds in mouse fungiform papillae consist of ,50 elongated cells (TBCs), where fewer than three TBCs have synaptic contacts with taste nerves. We investigated whether the non-innervated TBCs were chemosensitive using a voltage-sensitive dye, tetramethylrhodamine methyl ester (TMRM), under in situ optical recording conditions. 2Prior to the optical recordings, we investigated the magnitude and polarity of receptor potentials under in situ whole-cell clamp conditions. In response to 10 mM HCl, several TBCs were depolarized by ,25 mV and elicited action potentials, while other TBCs were hyperpolarized by ,12 mV. The TBCs eliciting hyperpolarizing receptor potentials also generated action potentials on electrical stimulation. 3A mixture of 100 mM NaCl, 10 mM HCl and 500 mM sucrose depolarized six TBCs and hyperpolarized another three TBCs out of 13 identified TBCs in a taste bud viewed by optical section. In an optical section of another taste bud, 1 M NaCl depolarized five TBCs and hyperpolarized another two TBCs out of 11 identified TBCs. 4The number of chemosensitive TBCs was much larger than the number of innervated TBCs in a taste bud, indicating the existence of chemosensitivity in non-innervated TBCs. There was a tendency for TBCs eliciting the same polarity of receptor potential to occur together in taste buds. We discuss the role of non-innervated TBCs in taste information processing. [source] | |||||||||||||