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Biomedical subjects

M W Berns

Publications and source records attributed to M W Berns.

At least 109 records · Page 6Linked to original sources
PubMed1992

Optical trapping in animal and fungal cells using a tunable, near-infrared titanium-sapphire laser.

We have compared two different laser-induced optical light traps for their utility in moving organelles within living animal cells and walled fungal cells. The first trap employed a continuous wave neodymium-yttrium aluminum garnet (Nd-YAG) laser at a wavelength of 1.06 micron. A second trap was constructed using a titanium-sapphire laser tunable from 700 to 1000 nm. With the latter trap we were able to achieve much stronger traps with less laser power and without damage to either mitochondria or spindles. Chromosomes and nuclei were easily displaced, nucleoli were separated and moved far away from interphase nuclei, and Woronin bodies were removed from septa. In comparison, these manipulations were not possible with the Nd-YAG laser-induced trap. The optical force trap induced by the tunable titanium-sapphire laser should find wide application in experimental cell biology because the wavelength can be selected for maximization of force production and minimization of energy absorption which leads to unwanted cell damage.

Aluminum Oxide↗
PubMed1992

Laser vs. suture nerve anastomosis.

The repair of injured or transected nerves is frequently encountered in head and neck surgery. Recently, CO2 lasers with milliwatt capability and micrometer spot sizes have been developed that may be used in nerve anastomosis. A comparative study was performed between microsuture and CO2 laser repair of transected sciatic nerves in rats. Nerve regeneration was measured in terms of morphology, electrophysiology, and function. Histologic studies revealed no difference in the size and number of regenerated axons, although there was less scar tissue formation at the anastomotic site with the laser repair. EMG and nerve conduction velocity were similar for the two repair methods. Functional recovery, as determined objectively using measurements of gait footprints, showed no difference between suture and laser repair. Laser-repaired nerves did have a higher dehiscence rate, although this problem can probably be prevented by splinting the rats postoperatively. However, laser repair was faster and simpler than suture repair and required less manipulation of the nerve. This study shows laser repair of peripheral nerves is possible with results comparable to conventional microsuture neurorrhaphy. Laser nerve anastomosis may be an effective alternative to suture nerve repair.

Anastomosis, Surgical↗
PubMed1992

Acute ultrastructural changes of cornea after excimer laser ablation.

Corneal ultrastructural changes induced by an argon fluoride excimer laser using different parameters were investigated. Twenty-eight rabbit corneas were ablated at light doses per pulse and repetition rates ranging from 25-800 mJ/cm2 and 1-100 Hz, respectively, at four different total light doses (25-150 J/cm2). Transmission electron microscopy showed that corneal ablations done at subthreshold light doses per pulse with repetition rates higher than 30 Hz and with an exposure more than 100 sec caused significant surface coagulation and an increase in pseudomembrane thickness. These changes were not observed in ablations done above threshold light doses per pulse, regardless of repetition rate and exposure time. However, repetition rates as high as 80 Hz caused damage to the endothelium and Descemet's membrane at the same ablation depth that did not cause such damage using repetition rates under 40 Hz. It appears that high repetition rates used during excimer laser corneal surgery may cause irreversible damage to the cornea.

Animals↗
PubMed1992

Ab-interno neodymium:YAG versus erbium:YAG laser sclerostomies in a rabbit model.

This study was undertaken to determine whether thermally-induced tissue necrosis was a factor in ab-interno contact-laser sclerostomy failure. A rabbit model was used to compare the continuous-wave Neodymium (Nd):YAG with the pulsed Erbium (Er):YAG laser with respect to such failure. Laser energy was focused into a fused-silica fiber optic (400 microns) for the Nd:YAG laser (12 W; 3 to 5 seconds), and into a single-crystal, uncladded sapphire fiber optic (250 microns) for the Er:YAG laser (7 to 8 mJ; 250 microseconds; 6 to 8 pulses). The Nd:YAG and Er:YAG lasers required from 21 to 35 J and from 42 to 64 mJ, respectively, to create the sclerostomies. Filtering blebs and intraocular pressure reduction lasted longer (log-rank test; P less than .03) and surgical complications were fewer in the Er:YAG group than in the Nd:YAG group. By creating sclerostomies with minimal thermal damage, the Er:YAG laser may offer significant clinical advantages over lasers producing larger thermal effects.

Animals↗
PubMed1991

Thoracoscopic carbon dioxide laser treatment of bullous emphysema.

A new technique of thoracoscopic laser ablation of pulmonary bullae suitable for patients with multiple bullae and diffuse emphysema was developed and assessed in 22 patients. 20 of 22 patients survived. Pre-operative and postoperative functional evaluation is available for the 11 patients followed up for more than a month; at 1 to 3 months postoperatively there were increases in FVC (mean 2.0 litres pre-operatively to 2.7 litres postoperatively, p less than 0.001), in FEV1 (0.74 to 1.06 litres, p = 0.01), and in maximum exercise treadmill times (5.4 min to 8.0 min, p less than 0.01). Postoperative air leaks lasted a mean of 13 days and usually resolved spontaneously. Other complications were bleeding (1 patient) and unilateral acute lung injury (1 patient). These results suggest that selected patients with diffuse emphysema and pulmonary bullae may benefit from thoracoscopic carbon dioxide laser ablation.

Cysts↗
PubMed1991

Photodynamic therapy of spontaneous cancers in felines, canines, and snakes with chloro-aluminum sulfonated phthalocyanine.

This is the first report on the photodynamic treatment with a second-generation sensitizer, chloro-aluminum sulfonated phthalocyanine (CASPc) of spontaneously arising tumors and on the photodynamic therapy (PDT) of snake neoplasms. Each of 10 cats, 2 dogs, and 3 snakes presenting with a variety of tumor types (squamous cell carcinoma, mast cell malignant tumor, and mixed carcinoma/sarcoma) was given an intravenous injection of 1 mg of CASPc per kilogram body weight 48 hours prior to irradiation with 675-nm light. Some larger tumors (greater than 1.5 cm deep) were surgically debulked prior to PDT. No significant systemic toxicity or skin photosensitization was observed in any animal. The tumor responses were comparable to those seen with conventional cryotherapy, hyperthermia, or surgery. PDT with CASPc of these cases led to 67% (12 of 18) complete response, 22% (4 of 18) partial response, and 11% (2 of 18) no response (less than 50% reduction in tumor size). Four cases could not be evaluated. Since the overall tumor response to CASPc is very good, and the problem of skin photosensitivity is nonexistent, it is expected that using CASPc-PDT to eradicate human tumors would also yield comparable results. Further studies with long-term follow-up are necessary to optimize the use of CASPc-PDT in veterinary and human medicine.

Animals↗
PubMed1991

Laser induced cell fusion in combination with optical tweezers: the laser cell fusion trap.

A single-beam gradient force optical trap was combined with a pulsed UV laser microbeam in order to perform laser induced cell fusion. This combination offers the possibility to selectively fuse two single cells without critical chemical or electrical treatment. The optical trap was created by directing a Nd:YAG laser, at a wavelength of 1.06 microns, into a microscope and focusing the laser beam with a high numerical aperture objective. The UV laser microbeam, produced by a nitrogen-pumped dye laser (366 nm), was collinear with the trapping beam. Once inside the trap, two cells could be fused with several pulses of the UV laser microbeam, attenuated to an energy of approximately 1 microJ/pulse in the object plane. This method of laser induced cell fusion should provide increased selectivity and efficiency in generating viable hybrid cells.

Animals↗
PubMed1991

Laser trabecular ablation (LTA).

As part of a pilot study for glaucoma surgery, the use of 3 infrared solid state lasers with 4 fiber optic delivery systems to ablate human trabecular meshwork was investigated. Laser trabecular ablation (LTA) was attempted with the Erbium:YAG (2.94 microns), Erbium:YSGG (2.79 microns), and Holmium:YSGG (2.1 microns) lasers. Laser energy was delivered as a single pulse (250 microseconds) by tissue fiber optic contact with low hydroxyl-fused silica (200 and 500 microns), zirconium fluoride (250 microns), or sapphire (250 microns) fiber optics. Total energy required and thermal effects decreased as laser wavelength increased. LTA was best achieved at 2.94 microns (4 mJ total energy; energy densities = 8.2-12.7 J/cm2; pulse length 250 microseconds) with average thermal damage zones of 5.3-10.3 +/- 1.3-2.4 microns (means +/- SDs) to contiguous structures. This finding has potential applications in the surgical treatment of open-angle and congenital glaucoma and may minimize failure rates seen in other types of surgery on the trabecular meshwork where disrupted trabecular meshwork is not removed.

Aluminum Oxide↗
PubMed1991

Selectivity, efficiency, and surface characteristics of hard dental tissues ablated with ArF pulsed excimer lasers.

Lasers are finding expanding applications in the field of dentistry. Cutting in soft tissue, hard dental material ablation, caries removal, and root canal therapy are only a few examples of dental laser uses. In this article, the application of short pulse ArF excimer laser to ablation of dentin and enamel is investigated. In particular, the effect of laser pulse repetition rates (PRR) and fluence levels on the efficiency of the ablation process and on the average thermal response of ablated surfaces is investigated. Ablation of dentin was found to be considerably more efficient than the ablation of enamel and depends exponentially on the laser fluence. Both dentin and enamel surfaces showed an increase in surface temperature with repetition rate. At lower PRR, however, temperature increases are very small. Surface temperature was also found to increase with laser fluence, although this increase is very small at laser PRR of 5 Hz or less. Tissue ablation rates were found to be comparable to or better than other nanosecond lasers, and left smooth surfaces, free of thermal damage. Microscopic examination of the ablated surface shows no crack formation, charring, discoloration, or any other thermal damage. The ablated surfaces appear to be very smooth, highly polished, and glossy looking as if they were subjected to thermal melting. This observation is indeed confirmed under scanning electron microscopy (SEM), where evidence of localized melting of the tissue is observed. Furthermore, a close SEM examination of the dentin surface reveals a selectively ablated intertubular dentin, while the remaining pillar-like dentin tubules are sealed off with fused peritubular dentin. At all fluence levels and PRR, the first three to four pulses impinging on an untreated enamel surface produced unusually large plumes of debris which were different in size, texture, and fluorescence emission characteristics from the ablation products of subsequent pulses. It is believed that these different ablation characteristics are a consequence of the pulsed ArF laser's ability to selectively remove residual matter from the more resistant enamel surface.

Acid Etching, Dental↗
PubMed1991

Micromanipulation of mitotic chromosomes in PTK2 cells using laser-induced optical forces ("optical tweezers").

To study the potential use of optical forces to manipulate chromosome movement, we have used a Nd:YAG laser at a wavelength of 1.06 microns focused into a phase contrast microscope. Metaphase and anaphase chromosomes were exposed while being monitored by video microscopy. The results indicated that when optical forces were applied to late-moving metaphase chromosomes on the side closest to the nearest spindle pole, the trapped chromosomes initiated movement to the metaphase plate. The chromosome velocities were two to eight times the normal rate depending on the chromosome size, geometry, and trapping site. At the initiation of anaphase, a pair of chromatids could be held by the optical trap and kept motionless throughout anaphase while the other pairs of chromatids separated and moved to opposite spindle poles. As a result, the trapped chromosome either was incorporated into one of the daughter cells or was lost in the cleavage furrow, or the two chromatids eventually separated and moved to their respective daughter cells. If the trap was removed at the beginning of anaphase B, the chromosome moved back to the poles. Our experiments demonstrate that the laser-induced optical force trap is a potential new technique to study noninvasively the mitotic spindle of living cells.

Analysis of Variance↗
PubMed1991

Activation of macrophages by Photofrin II during photodynamic therapy.

In order to obtain information about the activation of macrophages (M phi s) during photodynamic therapy (PDT), the influence of Photofrin II (Pf II) on the viability of thioglycollate-elicited murine M phi s and the subsequent generation of superoxide anion was studied. Irradiations were performed at an energy density of 5 J cm-2, a power density of 150 mW cm-2 and a wavelength of 405 nm. Viability of M phi s was assessed using the acridine orange-ethidium bromide assay. Superoxide anion generation was determined using ferricytochrome c (cyt c) and nitroblue tetrazolium (NBT) reduction. Our results indicate that the M phi s are highly susceptible to PDT as their viability is decreased to approximately 30% by 1 microgram ml-1 Pf II at the energy density indicated above. Within the first 30 min of addition of the photosensitizer, a reducing agent is generated intracellularly by the stimulation of the M phi s. An extracellular release of superoxide anion does not occur, as measured by the cyt c assay. Preincubation of the cells for 1 or 24 h with Pf II and a second challenge with phorbol myristate acetate (PMA) does not enhance the reduction of NBT. Thus, Pf II exerts an immediate effect on the M phi s which could be interpreted as a first step for subsequent reactions.

Animals↗
PubMed1991

Laser surgery.

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Animals↗
PubMed1991

Micromanipulation of gametes using laser microbeams.

Various microsurgical procedures at the cellular and subcellular levels using laser non-touch techniques are presented and reviewed. In these procedures, the beams of light, varying in their wavelength (range: 14 ns to continuous wave), were directed via microscopes towards the target area. Micromanipulation of human spermatozoa with a laser-generated optical trap enabled the assessment of possible effects on sperm motility and measurements of the relative force generated by each single spermatozoon. Furthermore, the optical trap also provides a new approach to the measurements of intracellular forces without physically touching the cell or its organelles, and inducing chromosome movement during cell division is also possible. Laser beams in a specific configuration are able to induce minimal superficial damage to the zona pellucida of oocytes from various species. This manipulation is aimed at increasing the fertilization rate following insemination with low-quality spermatozoa. Another intracellular application of the laser beams is the destruction of extra pronuclei in polyspermic fertilized human oocytes. These procedures require special equipment which is not commonly available. However, simpler devices may be developed if the advantages of this novel technology are demonstrated.

Fertilization↗
PubMed1991

Direct experimental evidence for the existence, structural basis and function of astral forces during anaphase B in vivo.

The existence, structural basis and function of astral forces that are active during anaphase B in the fungus, Nectria haematococca, were revealed by experiments performed on living cells. When one of the two asters of a mitotic apparatus was damaged, the entire mitotic apparatus migrated rapidly in the direction of the opposing astral forces, showing that the force that accelerated spindle pole body separation in earlier experiments is located in the asters. When a strong solution of the antimicrotubule drug, MBC, was applied at anaphase A, tubulin immunocytochemistry showed that both astral and spindle microtubules were destroyed completely in less than a minute. As a result, separation of the spindle pole bodies during anaphase B almost stopped. By contrast, disrupting only the spindle microtubules with a laser microbeam increased the rate of spindle pole body separation more than fourfold. Taken together, these two experiments show that the astral forces are microtubule-dependent. When only one of the two or three bundles of spindle microtubules was broken at very early anaphase B, most such diminished spindles elongated at a normal rate, whereas others elongated at an increased rate. This result suggests that only a critical mass or number of spindle microtubules needs be present for the rate of spindle elongation to be fully governed, and that astral forces can accelerate the elongation of a weakened or diminished spindle.

Anaphase↗
PubMed1991

Excimer laser radiation for endarterectomy of experimental atheromas.

Open laser endarterectomy produces a smooth arterial surface with welded distal end points. This report evaluates 308-nm excimer laser radiation for the laser endarterectomy operation. Arteriosclerotic New Zealand white rabbits (N = 15) were studied. A thoraco-abdominal exploration was performed, the aorta was isolated, heparin was administered, and multiple endarterectomies were performed in each rabbit. A line of laser craters was created at the proximal and distal ends of an atheroma. Laser radiation was used to connect the craters to form the proximal and distal end points. The atheromas were dissected from the aorta with laser light and the end points were fused. The aortas were removed for light and electron microscopy and the animals were sacrificed. Excimer radiation was delivered by a 600-microns fiber at 50 mJ/pulse, 120-ns pulses and either 15- or 20-Hz frequency. At 15 Hz excimer laser endarterectomies showed no perforations along the surface or at the end points. The surfaces were smooth but the end points were not welded in place. At 20 Hz, perforations were seen along 7/11 surfaces and at 5/11 end points. Excimer laser endarterectomy is best performed at 15 Hz. The end points, however, cannot be welded with excimer laser radiation.

Animals↗