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.

Axial trapping forces exerted on microspheres are predicted using a Gaussian beam electromagnetic field model and a ray-optics model, and compared with experimental measurements. Ray-optics predicts a maximum trapping efficiency Q= -0.14 for optically trapped polystyrene microspheres in water, compared to a measured value of -0.12 ± 0.014 for 10 μm diam microspheres. When the microspheres are composed of amorphous silica, the predicted ray-optics Q decreases to -0.11, compared to a Q = -0.034 predicted by the electromagnetic field model, and a measured value of -0.012 ± 0.001 for 1 μm diam microspheres. These results indicate that the two models have applicability in two different size regimes, and thus, are complementary.

Optical traps offer the promise of being used as noninvasive micromanipulators for biological objects. We have developed an analytical model that accurately describes the forces exerted on dielectric microspheres while in a single-beam gradient force optical trap. The model can be extended to the trapping of biological objects. The model predicts the existence of a stable trapping point and an effective trapping range. A minimum trapping power of — 5 mW and an effective trapping range of 2.4 um have been measured for 10 um diameter dielectric microspheres, and are in reasonable agreement with expected results. In cell biology, we have used the optical trap to alter the movement of chromosomes within mitotic cells in vitro and to hold motile sperm cells. Results for the mitotic cells indicate that chromosome movement was initiated in the direction opposite to that of the applied force. Chromosome velocities as high as 48 um/min were observed, and are 24 times faster than normal during cell division. In sperm trapping experiments, mean sperm velocities were unchanged following short (< 30 s) exposures in the trap, while longer exposures resulted in a decrease in mean sperm velocities. Given its noninvasive nature, optical traps should prove to be useful tools in the study of biological processes. © 1990 IEEE

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.

A pulsed-laser microbeam at 532 nm wavelength (optical scissors) and a laser-induced optical trap at 1064 nm wavelength (optical tweezers) have been successively combined to dissect and manipulate chromosomes in live newt lung epithelial cells. These preliminary experimental results demonstrated that chromosome fragments dissected by laser microbeam surgery, regardless of their size, could be easily pulled or rotated by optical forces when positioned at the periphery of the mitotic spindle. In addition, chromosome arms which were not subjected to laser microsurgery also could be moved with the optical tweezers at the spindle periphery. In our previous study on rat kangaroo kidney cells (PTK2), this degree of facilty in manipulating chromosome movement was not possible, most likely due to the close proximity of the intermediate filament "cage" to the spindle. It is concluded herein that optical scissors and tweezers can be used in combination to study the interaction of chromosomes with the mitotic spindle in cells where the peripheral regions of the spindle are unobstructed by intermediate filaments. This can be performed on newt cells, where the diameter of the cage can be substantially larger than the diameter of the spindle.