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Biomedical subjects

W H Wright

Publications and source records attributed to W H Wright.

At least 19 recordsLinked 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↗
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

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

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

Mid-infrared laser ablation of stratum corneum enhances in vitro percutaneous transport of drugs.

The precise removal of stratum corneum from cadaveric swine skin by a mid-infrared erbium:yttrium scandium gallium garnet laser (lambda = 2.79 microns; 250 microseconds pulse width) was assessed by electrical resistance measurements and documented by histology. The effects of stratum corneum removal by laser ablation and by adhesive tape-stripping on the in vitro penetration of 3H-hydrocortisone and 125I-gamma-interferon were determined. Excised swine skin was irradiated with laser (1 J/cm2; 31 mJ/pulse; 1 Hz; 2 mm spot diameter). For skin penetration studies, laser pulses were delivered to discrete 2-mm areas to ablate up to 12.6% of the total 3-cm2 stratum corneum diffusional area. Franz in vitro skin penetration chambers were used to measure the cumulative 48-h penetration of 3H-hydrocortisone and 125I-gamma-interferon in laser-treated and tape-stripped skin. Electrical resistance measurements and histologic studies demonstrated that 10-14 laser pulses at the above energy density were required to abolish skin resistance and selectively ablate stratum corneum without damage to adjacent dermal structures. Laser ablation of 12.6% of the surface area of stratum corneum produced a 2.8 and 2.1-times increase in permeability constant (kp) for 3H-hydrocortisone and 125I-gamma-interferon, respectively. These studies demonstrate that a pulsed mid-infrared laser can reliably and precisely remove the stratum corneum, facilitating penetration of large molecules such as 125I-gamma-interferon that cannot penetrate intact skin. This new technique may be useful for basic and clinical investigation of skin barrier properties.

Administration, Cutaneous↗
PubMed1990

Force generated by human sperm correlated to velocity and determined using a laser generated optical trap.

The development of the single beam gradient force optical trap has made it possible to manipulate cells solely by laser light. A continuous wave Nd:YAG (1.06 microns) laser beam was directed into a conventional microscope and focused onto the viewing plane by the objective lens. The laser beam power at which human sperm were released from the trap was measured and correlated to the sperm's linear velocity before trapping. The mean trapping power readings for slow, medium, and fast motile sperm were 57, 73, and 84 mW, respectively. The analysis of measurements over the total population demonstrated that zig-zag motile sperm had significantly higher mean power readings when compared with straight motile sperm with similar mean linear velocities. In two cases, specimens required significantly less trapping power when the measurements were repeated 24 hours later.

Ejaculation↗
PubMed1989

Use of a laser-induced optical force trap to study chromosome movement on the mitotic spindle.

A laser-induced optical force trap was used to alter the movement of chromosomes in mitotic cells in vitro. The trap was produced by using a 1.06-microns neodymium YAG (yttrium/aluminum garnet) laser focused through a phase-contrast microscope. The trap was applied to one side of centrophilic chromosomes off the mitotic spindle and to late-moving chromosomes on the mitotic spindle. In both situations, chromosome movement was initiated in the direction opposite to that of the applied force. When the force was applied, chromosomes moved at velocities 10-20 times normal. These studies verify and extend the feasibility of using this new technique to study factors that influence organelle motility.

Animals↗
PubMed1989

Micromanipulation of sperm by a laser generated optical trap.

The force generated by the radiation pressure of a low power laser beam induces an optical trap which may be used to manipulate sperm. We studied the effect of the optical trap on sperm motility. A Nd:YAG laser beam was coupled to a conventional microscope and focused into the viewing plane by the objective lens. Sperm were caught in the trap and manipulated by a joy stick controlled motorized stage. After different exposure periods, the velocity and patterns were analysed by a computerized image processor. There were minor changes in sperm velocity when exposed to the trap for 30 seconds or less. A gradual decrease in the mean linear velocity was observed after 45 seconds of exposure. This optical micromanipulator may also be useful for studying the force generated by a single spermatozoa and evaluating the influence of drugs on motility.

Humans↗
PubMed1988

Combination cytotoxic chemotherapy with cisplatin or doxorubicin and photodynamic therapy in murine tumors.

This study was designed to evaluate the interaction of photodynamic therapy (PDT) and chemotherapy in an animal model. PDT is based on the interaction of hematoporphyrin derivative and red light of the appropriate wavelength (630 nm) and intensity. Two tumor models were utilized: C3H/Km mice bearing the RIF-1 tumor and BALB/c mice bearing the EMT-6 tumor. Tumor-bearing mice were treated with either cisplatin (DDP), doxorubicin (ADM), PDT, or a combination of drug and PDT. It was demonstrated that the RIF-1 tumor was sensitive to DDP and insensitive to both PDT and ADM. There was no additional antitumor effect when either drug was combined with PDT. The EMT-6 tumor was moderately sensitive to PDT and mildly sensitive to both DDP and ADM. Although the addition of DDP did not potentiate tumor destruction, the addition of ADM significantly enhanced the effect of PDT (P = .01). The enhanced activity of the combination of PDT and ADM appeared to be the result of increased activity of ADM alone, when illuminated with red (630 nm) light. This potentiation may be due to a photochemical process or may be secondary to the mild hyperthermia generated by illumination with the laser. This study demonstrates that PDT combined with cytotoxic chemotherapy is well tolerated in these animals and that certain combinations of PDT and chemotherapy may result in an enhanced tumoricidal effect.

Animals↗
PubMed1988

Ablation of bone and methacrylate by a prototype mid-infrared erbium:YAG laser.

An erbium:YAG laser was used to generate 200-microseconds pulses of mid-infrared 2.94-microns light in both the single and multimode configurations. Laser pulses were focused on the surfaces of both rabbit long bones and methacrylate blocks, and the tissue response was examined histologically. The depth of thermal injury was determined by ocular micrometry. Over all energy levels tested, the erbium:YAG laser produced ablation of bone and methacrylate with minimal thermal damage to adjacent tissue. Increasing the laser energy per pulse produced increasingly wider and deeper grooves in both bone and methacrylate. However, such increase in laser energy produced a proportionately greater increase in the zone of thermal injury in methacrylate as compared with bone. These studies suggest the feasibility of a surgical erbium:YAG laser in orthopaedics and other forms of ablative surgery.

Animals↗
PubMed1985

Comparison of fluorescence and photodynamic activities of whole hematoporphyrin derivative and its enriched active components.

The in vivo biologic activities of the hematoporphyrin derivative (Photofrin) and the enriched, so-called "active fraction" (Photofrin II) were determined by measuring the necrosis produced in implanted tumors in DBA/2Ha mice exposed to various total doses of light (20-100 J/cm2) after ip administration of 10 mg/kg standard doses of either Photofrin or Photofrin II. Total relative percentage increase in fluorescence in tumor tissue, as compared to fluorescence in control tissue, also was measured for both Photofrin and Photofrin II. In response to total light doses (630 nm) of 40-100 J/cm2, mice that received Photofrin had comparable amounts of tumor necrosis to those mice that received Photofrin II. At doses of 40-60 J/cm2, 80% tumor destruction resulted, and at 80-100 J/cm2, tumor destruction was 100%. However, at a total light dose of 20 J/cm2, the tumors that received Photofrin II exhibited 60-80% tumor necrosis, whereas those animals that received Photofrin had only small areas of patchy necrosis associated with signs of vascular thrombosis and hemorrhage into the surrounding perivascular stroma. A 25.2% total increase in maximal tissue fluorescence over that in controls was observed for animals that received Photofrin II, as compared to 13.9% for those animals that received Photofrin. It is concluded that the greater demonstrable efficacy of treatment with Photofrin II, as compared to treatment with Photofrin, is due to enrichment of those nonpolar hydrophobic components of the hematoporphyrin derivative mixture that are thought to be primarily responsible for the in vivo biologic activities.

Animals↗
PubMed1985

Histopathological comparison of the effects of hematoporphyrin derivative on two different murine tumors using computer-enhanced digital video fluorescence microscopy.

A comparison study of the effects of hematoporphyrin derivative (HPD) photoradiation therapy on two different mouse sarcoma tumor model systems (RIF-1 + EMT-6) was performed. Twenty-four h after i.v. administration of HPD, the responses to total laser light doses of 50-400 J/cm2 were evaluated by histological examination and the uptake and distribution of HPD using a computer enhanced digital video fluorescence microscopy technique. In response to total laser light dose (630 nm) of 50-400 J/cm2, 40 mice with RIF-1 tumor showed only minimal superficial tumor necrosis upon histological examination and a 9-12% increase in maximal tissue fluorescence. In contrast, 40 mice with EMT-6 tumor showed marked areas of patchy coagulation necrosis and vascular hemorrhage at doses as low as 50 J/cm2 and essentially total tumor destruction at total light doses of 150 J/cm2 or more. A 59-74% increase in maximal tissue fluorescence was observed using digital video fluorescence microscopy. It is concluded that the greater efficacy of treatment in the EMT-6 tumor as compared to the RIF-1 tumor was due to the greater localization of HPD as demonstrated by digital video fluorescence microscopy.

Animals↗
PubMed1978

The relationship between attachment height and attachment area of teeth using a digitizer and a digital computer.

Nine lower first bicuspid teeth were mounted in blocks of plastic and their roots were serially sectioned and photographed. Data from the photographs were compiled by a digitizer and a digital computer, which calculated the cumulative root surface area for each millimeter of root height. From these data, three variables, which relate root surface area to root height, were calculated. It was found that if one-half of the height of attachment to the root were lost due to periodontal disease, a mean of 61.5% of the actual attachment area to the root is lost. If a mean of 5.72 mm of root attachment height is lost, or if a mean of 60.6% of the same root height remains, only one-half of the total root attachment area remains to provide tooth support.

Bicuspid↗