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[Retinal photocoagulation with a pulsed, frequency-doubled Nd:YAG laser (532 nm)].

The small difference in wavelength between an argon laser (514 nm) and a frequency-doubled Nd:YAG laser (532 nm), together with the advantage of the solid-state technology, makes the Nd:YAG laser likely to play a major role in retinal photocoagulation in the near future. For technical reasons all frequency-doubled Nd:YAG lasers work in a quasi-continuous mode, emitting a burst of highly repetitive short laser pulses during the exposure time desired. We investigated the side effects due to high peak irradiances of those short laser pulse trains (Crystal Focus Nd:YAG laser, Emerald; pulse duration 1-10 microseconds, repetition rate 13 KHz) in rabbits in comparison with a standard argon laser system (Zeiss, Visulas, Argon II). The energy necessary for blanching the retina was similar in both cases. As opposed to the argon laser system, subretinal bubbles were regularly visible ophthalmoscopically with the Nd:YAG system, when average powers as high as 200 mW were used. The ED50 power for bubble formation is about 2-3 times above the ED50 power for blanching. Thermal calculations show that this bubble formation effect is likely to be related to the peak power of the short pulses. The hemorrhage threshold is similar in both systems. However, light microscopically there is no difference between the two laser systems. Panretinal photocoagulation (300-500 microns, 100-200 ms) in patients with proliferative diabetic retinopathy produced such bubbles about once per 1000 lesions.

Animals↗

Comparison of the erbium-yttrium aluminum garnet and carbon dioxide lasers for in vitro bone and cartilage ablation.

The in vitro bone- and cartilage-ablation characteristics of the solid-state erbium:yttrium aluminum garnet laser were compared to those of the carbon dioxide laser. Ablations of fresh, frozen cadaver septal cartilage and maxillary sinus bone were performed using total energies between 1 and 6 J. Specimens were studied using hematoxylin and eosin stain and digitized, computer-assisted measurements of 35-mm photographs. Erbium-yttrium aluminum garnet-ablated bone averaged 5 microns of adjacent tissue thermal injury, compared with 67 microns with carbon dioxide-ablated bone. Erbium-yttrium aluminum garnet-ablated cartilage averaged 2 microns of adjacent tissue thermal injury, compared with 21 microns with the carbon dioxide-ablated cartilage. The tissue-ablation characteristics of the erbium-yttrium aluminum garnet laser are promising for future otolaryngologic applications.

Aluminum↗

Fluorescence lifetime imaging microscopy: homodyne technique using high-speed gated image intensifier.

In the previous sections we demonstrated imaging of intracellular Ca2+ using our approach to FLIM. What other analytes can be imaged using FLIM? We have now characterized the lifetime of a good number of ion indicators. Based on these studies we know that Cl- can be imaged using FLIM with probes such as SPQ or MQAE, pH can be imaged using resorufin and probes of the SNAFL and SNARF (Molecular Probes) series, and Mg2+ can be imaged using Magnesium Green, Mag-quin-2, or Mag-quin-1 (Molecular Probes). At present, the probe for K+, as PBFI, are just adequate as a lifetime probe, but it seems likely that newer probes for Na+ (Sodium Green) and K+ will be practical for effective imaging. Of course, imaging of oxygen is possible using a wide variety of fluorophores. It should be noted that a wide variety of substances and/or phenomena are known to alter decay times, acting as quenchers. These include the phenomena of resonance energy transfer, collisional quenching, temperature effects, and viscosity effects. Also, the FLIM method is not limited to microscopic objects but can be possibly used in remote imaging of any object. Hence, FLIM will allow the imaging of the chemical and physical properties of objects based on the effects of the local environment on the decay kinetics of fluorophores. The instrumentation for FLIM is presently complex and requires a moderately complex laser source, a gain-modulated image intensifier, and a slow-scan CCD camera. However, one can readily imagine the instrumentation becoming rather compact, and even all solid-state, owing to advances in laser and CCD technologies and, more importantly, advances in probe chemistry. To be specific, the dye laser shown in Fig. 1 may be replaced by a simpler UV laser, such as the 354 nm HeCd laser which has become available (Fig. 11). Intensity modulation of a continuous wave sources can be accomplished with acoustooptic modulators. The scientific slow-scan CCD cameras are presently rather expensive, but they are used in the present instrumentation because of their linearity and high dynamic range. However, the increasing use of CCD detectors suggest that even the scientific-grade CCD cameras will soon become less costly. Additionally, the frame rates of these detectors continue to increase in response to the need for faster imaging. Furthermore, the performance of the video CCD cameras is increasing, as seen by the introduction of 10-bit video analog-to-digital (A/D) converters.(ABSTRACT TRUNCATED AT 400 WORDS)

Aminoquinolines↗

Laser scleral buckling for retinal reattachment.

The holmium:yttrium aluminum garnet (Ho:YAG) laser produces infrared light that is absorbed inside scleral tissues, which increases local temperature, thereby shrinking collagen and inducing a buckle effect. Scleral indentation was induced in ten cadaver eyes by shrinkage of scleral collagen fibers by using a pulsed solid-state Ho:YAG (2.1-microns) laser with fiberoptic delivery. The amount of laser-induced buckling effect is controlled by selecting laser treatment settings such as beam spot, radiant exposure, and number of pulses. With treatment using 11.3 +/- 1.2 J/cm2 of laser radiant exposure and five pulses, laser-induced scleral shrinkage affected only the external two thirds of the scleral tissue. No thermal damage or disruption was observed in subjacent retinal pigment epithelium, choroid, or retina. The coupling of two appropriately selected lasers may allow laser-induced scleral buckling and transscleral retinal photocoagulation by using the same laser probe for retinal reattachment.

Collagen↗

Holmium: YAG laser-assisted coronary angioplasty with multifiber delivery catheters.

Mid-infrared lasers are attractive for coronary angioplasty based upon their excellent fiberoptic transmission and enhanced tissue absorption. Using a solid-state, mid-infrared holmium:YAG laser with prototype multifiber laser delivery catheters, we performed coronary laser angioplasty with or without adjunctive balloon angioplasty or directional atherectomy in 14 patients with 17 stenoses. Procedural laser success was obtained in 13/14 (93%) patients and 16/17 (94%) lesions; however clinical success was achieved in 9/14 (64%) patients and 12/17 (71%) stenoses. Angiographic restenosis at 4.6 +/- 1.6 months was found in 5/8 (63%) patients. We achieved an excellent procedural laser success rate in patients with generally unfavorable angioplasty anatomy. However, our clinical success rate was not different from that expected with conventional angioplasty alone. The holmium laser remains an attractive energy source for laser angioplasty; but as is the case for all coronary laser systems, its utility as a stand-alone therapy is limited by catheters which create small channels. In this small group, we could demonstrate no clinical benefit for laser-assisted angioplasty in complex coronary lesions. Our results suggest that a randomized trial comparing laser-assisted angioplasty and conventional angioplasty be performed to determine the clinical benefits of this more expensive therapy.

Adult↗

[Possible applications of laser in otorhinolaryngology].

Laser therapy in otolaryngology--head and neck surgery using laser types working in the range of the visible and infrared part of the electromagnetic spectrum is well established. The laser types usually employed for micro- and macrosurgery of the aero-digestive tract and for microsurgery of the middle ear, involving thermal ablation of tissue and extensive areas of damage are described, as well as recent advances in laser technology for future use. The mechanisms by which non-thermal, photo-ablative pulsed solid-state crystal infrared and UV-excimer lasers and photodynamic therapy affect biological tissues are reported. The advantageous main areas of application of the different types of laser according to their physical properties are compared on the basis of the results of our own studies and the relevant literature.

Animals↗

Posterior capsule polishing with the neodymium:YLF picosecond laser: model eye study.

PURPOSE: To determine the ability and threshold energy of the neodymium:yttrium-lithium-fluoride (Nd:YLF) picosecond laser to achieve micron-level polishing of a latex posterior capsule facsimile (PCF) as an alternative to laser capsulotomy to treat posterior capsule opacification. SETTING: University of Rochester Medical Center, Rochester, New York, USA. METHODS: A solid-state, mode-locked Nd:YLF picosecond laser was used to polish a latex PCF in contact with a poly(methyl methacrylate) intraocular lens (IOL) in an experimental model eye. Eight study groups were treated at different energy levels ranging from 5 to 15 microJ. All treatments were done at least three times in different latex capsules and lenses. An atomic force microscope was used to measure IOL damage and an interferometric surface analysis microscope to assess the polishing effect on the PCF. The IOLs were further subjected to a scatter analysis to assess the optical significance of the damage produced. RESULTS: The latex PCF revealed a polishing effect with all energy settings used. The IOLs were damaged with all energy settings but 5 microJ. Energy settings higher than 5 microJ caused significantly more polishing effect to the latex and damage to the lenses. At the 10 microJ energy level, a single parameter with no depth produced a relative polishing depth of 3.01 microns +/- 0.10 (root mean square +/- SD). At this energy, the damage to the IOLs was 188 +/- 20.52 nm, and it was associated with typical craters over the surface at regular intervals that corresponded to each individual laser pulse. CONCLUSION: This model documented the feasibility of achieving micron-level precision in excising material with the picosecond laser and showed that posterior capsule polishing should be feasible and safe in human eyes.

Humans↗

Laser facilitated angioplasty and thrombolysis in acute myocardial infarction complicated by prolonged or recurrent chest pain.

To date, application of laser angioplasty in acute myocardial infarction (MI) has not been reported. In nine patients with acute myocardial infarction complicated by continuous or recurrent severe ischemia and chest pain, a mid-infrared, solid-state, pulse-wave holmium/thulium:YAG coronary laser was applied. In six of these patients the laser was specifically utilized for the purpose of coronary thrombolysis. In each case a guidewire was placed across the stenosis and a multifiber laser catheter was utilized, emitting 250-600 mJ/pulse at 5 Hz, followed by adjunctive balloon angioplasty. Laser success (defined as ability to cross the lesion, reduction of > or = 20% in stenosis and thrombolysis when a thrombus is present) was achieved in all patients. Final angiograms revealed residual stenosis < or = 30%, adequate thrombolysis and no major complication (MI, perforation, emergency CABGS, CVA, death) in each patient. Clinically, all nine patients improved, survived the acute infarction and were discharged. This initial clinical experience demonstrates the feasibility and safety of holmium/thulium:YAG laser application in thrombolysis and plaque ablation in selected patients who experience acute myocardial infarction complicated by prolonged or recurrent ischemia and chest pain.

Adult↗

Lithotripsy with a Q-switched alexandrite laser system. Preliminary in vivo and in vitro results.

An alexandrite laser system which emits a wavelength of 750 nm, has recently been proposed as a substitute for the pulsed-dye laser (504 nm) for laser lithotripsy. We have carried out in vitro lithotripsy trials in order to evaluate the efficiency of a flashlamp pumped Q-switched alexandrite laser. In the first experiment, we compared the Q-switched alexandrite laser to the pulsed-dye laser with respect to their effectiveness in fragmentizing urinary and model stones (HMT target stones). In the second experiment, we evaluated the effect of Q-switched alexandrite radiation on the urinary wall (bladder, ureter, renal pelvis and liver) of animals (Wistar rats and rabbits) under general anesthesia. In conclusion, the alexandrite laser system is comparable to the pulsed dye laser. The 750-nm wavelength involves minimal risk of damage to the urinary tract and constitutes a safe alternative, especially during blind lithotripsy. It is a compact solid-state system, with no toxic chemical agents as laser material.

Animals↗

THC:YAG nasolacrimal duct recanalization.

In an early exploration of a new technique for creating a patent nasolacrimal duct system, a chromium-sensitized and thulium- and holmium-doped YAG laser was used to canalize the nasolacrimal duct of a fresh-frozen bisected human cadaver head. The laser--long-pulsed (300 milliseconds), compact, self-contained, and solid-state--operates in the near infrared range (2.1 microns). The technique involved passing a 550-micrometer quartz fiberoptic through the dilated superior punctum and canalicular system, down through the nasolacrimal duct. The quartz fiberoptic was then withdrawn into the area of the lacrimal sac. Pulse energies of 500 mJ were used at a repetition rate of five pulses per second to ablate the duct lining. The nasolacrimal duct was then unroofed and the epithelial lining of the duct histopathologically evaluated.

Dacryocystorhinostomy↗

Chiral detection in high-performance liquid chromatography by vibrational circular dichroism.

A novel chiral detector for high-performance liquid chromatography has been developed. This detector is based on the measurement of circular dichroism of chiral effluents in the infrared region, i.e., vibrational circular dichroism (VCD). In this instrument, a solid-state spectral tunable (from 2.4 to 3.5 microns) F-center laser was used as the light source. The linearly polarized laser beam was converted into left circularly polarized light (LCPL) and right circularly polarized light (RCPL) at 42 kHz by means of a photoelastic modulator. The intensity of the LCPL and RCPL transmitted through the sample was measured by a liquid nitrogen cooled indium antimonide detector. Double modulation was employed to reduce the noise associated with the laser beam. Specifically, the linearly polarized laser beam, prior to being converted to CPL, was modulated at 85 Hz by a mechanical chopper. Demodulation and amplification were accomplished with the use of two lock-in amplifiers. In its present configuration, the instrument can be used to measure the VCD of O-H groups. Its sensitivity is so high that it was able, for the first time, to detect chirally (with limits of detection of micrograms) (R)- and (S)-2,2,2-trifluoro-1-(9- anthryl)ethanol and (R)- and (S)-benzoin when these compounds were chromatographically separated from the corresponding racemic mixtures by a Chiralcel-OD column. The main advantage of this chiral detector is, however, its universality; i.e., it can be used to virtually detect any chiral compounds which has O-H group (e.g, aliphatic alcohols such as 2-octanol).

Chromatography, High Pressure Liquid↗

Fiber bundle based scanning detection system for automated DNA sequencing.

High-throughput DNA sequencing techniques are under rapid development currently, mainly triggered by the Human Genome Project. At the present time, slab gel based automated DNA sequencing is the standard procedure, utilizing fluorophore labeling and laser-induced fluorescence detection with scanning technology. In this paper, a novel, fiber-optic bundle based detection system is introduced, where a central illuminating fiber is used for the excitation of the electrophoretically separated fluorophore-labeled DNA sequencing fragments, along with several collecting fibers disposed around the illuminating fiber to collect the emitted fluorescent signal. As a model system, Cy5-labeled DNA sequencing fragments were separated on an ultrathin polyacrylamide slab gel and detected by the fiber bundle based laser-induced fluorescence detection system. A 640-nm diode laser was used to generate the illumination beam, and the emitted light collected by the fiber bundle was detected by a solid-state avalanche photodiode.

Autoanalysis↗

Surface selective 1H/29Si CP NMR by NOE enhancement from laser polarized xenon

The surface proton spin polarization created by the spin-polarization-induced nuclear Overhauser effect from optically polarized xenon can be transferred in a subsequent step by solid-state cross polarization to another nuclear spin species such as 29Si. The technique exploits the dipolar interactions of xenon nuclear spins with high gamma nuclei such as 1H, and is experimentally simpler than direct polarization transfer from 129Xe to heteronuclei such as 13C and 29Si. Copyright 1998 Academic Press.

Journal Article↗

Systemic nitric oxide synthase inhibition does not affect brain oxygenation during cortical spreading depression in rats: a noninvasive near-infrared spectroscopy and laser-Doppler flowmetry study.

Cortical spreading depression (CSD) has been implicated in the migraine aura and in stroke. This study demonstrates near-infrared spectroscopy (NIRS) for the first time as capable of noninvasive on-line detection of CSD in the pentobarbital-anesthetized rat. CSD was accompanied by a brief and rapid increase of regional CBF (by laser-Doppler flowmetry) to 200-400% baseline. NIRS demonstrates that this hyperperfusion is associated with concentration increases of oxyhemoglobin, while deoxyhemoglobin decreases. Simultaneously, oxygen partial pressure, measured on the brain surface with a solid-state polarographic probe, was shown to be raised by at least 14 mm Hg during CSD. Oxygen-dependent phosphorescence life-time quenching measurements confirmed this finding. NIRS data on cytochrome aa3, however, showed a CSD-related shift toward a more reduced state, despite raised blood oxygenation. This may suggest either limited O2 transport from the blood to mitochondria or decreased oxygen utilization during CSD as supposed by theories about compartmentalization of energy metabolism favoring glycolytic rather than aerobic energy supply during CSD. However, the data on cytochrome aa3 warrant caution and are discussed critically. Nitric oxide synthase inhibition by systemic application of N'-nitro-L-arginine had no significant effect on the perfusion response or the tissue PO2 during CSD. During most CSD episodes, a brief decrease in MABP by 4-8 mm Hg was noted that might be caused by functional decortication during CSD.

Animals↗

Diffusion-induced Ramsey narrowing.

Diffusion-induced Ramsey narrowing is characterized and identified as a general phenomenon, in which diffusion of coherence in and out of an interaction region such as a laser beam induces spectral narrowing of the associated resonance line shape. Illustrative experiments and an intuitive analytical model are presented for this spectral narrowing effect, which occurs commonly in optically interrogated atomic systems and may also be relevant to quantum dots and other solid-state spin systems.

Journal Article↗

Protein stability and conformational rearrangements in lipid bilayers: linear gramicidin, a model system.

The replacement of four tryptophans in gramicidin A by four phenylalanines (gramicidin M) causes no change in the molecular fold of this dimeric peptide in a low dielectric isotropic organic solvent, but the molecular folds are dramatically different in a lipid bilayer environment. The indoles of gramicidin A interact with the anisotropic bilayer environment to induce a change in the molecular fold. The double-helical fold of gramicidin M, as opposed to the single-stranded structure of gramicidin A, is not compatible with ion conductance. Gramicidin A/gramicidin M hybrid structures have also been prepared, and like gramicidin M homodimers, these dimeric hybrids appear to have a double-helical fold, suggesting that a couple of indoles are being buried in the bilayer interstices. To achieve this equilibrium structure (i.e., minimum energy conformation), incubation at 68 degrees C for 2 days is required. Kinetically trapped metastable structures may be more common in lipid bilayers than in an aqueous isotropic environment. Structural characterizations in the bilayers were achieved with solid-state NMR-derived orientational constraints from uniformly aligned lipid bilayer samples, and characterizations in organic solvents were accomplished by solution NMR.

Dimerization↗

Consideration of conformational transitions and racemization during process development of recombinant glucagon-like peptide-1.

Physicochemical characterization of dry, excipient-free recombinant glucagon-like peptide-1 (rGLP-1) indicates the conformation and purity of the bulk peptide is dependent on the purification scheme and the in-process storage and handling. The recombinant peptide preparations were highly pure and consistent with the expected primary structure and bioactivity. However, variations in solubility were observed for preparations processed by different methods. The differences in solubility were shown to be due to conformational differences induced during purification. A processing scheme was identified to produce rGLP-1 in its native, soluble form, which exhibits FT-IR spectra, consistent with glucagon-like peptide-1 synthesized by solid-state peptide synthesis. rGLP-1 was also found to undergo base-catalyzed amino acid racemization. Racemization can impact the yield and impurity profile of bulk rGLP-1, since the peptide is exposed to alkali during its purification. A combination of enzymatic digestion using leucine aminopeptidase (which cleaves N-terminal L-amino acids >> D-amino acids) and matrix-assisted laser desorption ionization mass spectrometry was used to identify racemization as a degradation pathway. The racemization rate increased with increasing temperature and base concentration, but decreased with increasing peptide concentration. The racemized peptides were shown to be less bioactive than rGLP-1.

Chemical Precipitation↗

Long-lifetime metal-ligand pH probe.

We describe the synthesis and fluorescence spectral characterization of a pH-sensitive metal-ligand complex, [Ru(deabpy)(bpy)2]2., where deabpy is 4,4'-diethylaminomethyl-2,2'-bipyridine. This metal-ligand complex (MLC) was found to display pH-dependent intensities, emission spectra, and decay times, with the changes centered near the physiological useful pH value of 7.5. The apparent pKa values were not found to be dependent on ionic strength. The compound was found to be useful for lifetime-based sensing by phase-modulation fluorometry. Global analysis of the intensity decays over a range of pH values revealed two decay times of 235 and 380 ns, associated with the protonated and unprotonated forms, respectively. Because of its long decay time, optical pH measurements could be accomplished by phase-modulation fluorometry with a conveniently low modulation frequency of 700 kHz. The lifetime data were obtained with either a amplitude-modulated laser or with an amplitude-modulated blue-light-emitting diode. This pH-sensitive complex also displays a modest spectral shift with change in pH, allowing its use as a wavelength-ratiometric MLC probe. One can imagine lifetime sensors for a variety of blood cations and point-of-care assays based on long-lifetime metal-ligand complexes and simple solid-state light sources and detectors.

2,2'-Dipyridyl↗