Design of computer-generated hologram with ring focus for nonmechanical corneal trephination with Er:YAG laser in penetrating keratoplasty.

PURPOSE: To calculate a beam-shaping optical element for homogeneous intensity distribution within a focal ring to be used in nonmechanical trephination with the Er:YAG laser in penetrating keratoplasty instead of a spot guiding device. METHODS: The phase distribution behind a holographic optical element (HOE) k psi(u) can be described by the addition of the hologram phase phiH(u) to the beam phase phiE(u): k psi(u) = phiH(u) + phiE(u), k = 2pi/lambda, where u denotes the coordinates inside the hologram aperture, k an integer, and lambda the laser wavelength. To avoid discontinuous wavefronts leading to speckle noise, a smooth phase function is necessary. After transforming the hologram aperture coordinates into the focal plane x in a focal distance f, psi can be retrieved from the slope equation: inverted delta psi(u) = x(u) - u/f. RESULTS: Creating a ring focus can be reduced to an essentially one-dimensional problem by separation of variables due to the symmetry condition. We calculated a computer-generated eight-level phase-only HOE with 4096 x 4096 pixels from a Gaussian-distributed 2.94 Er:YAG laser spot with a beam diameter of 10 mm and a focal distance of 100 mm. Thereby, a ring focus with an inner/outer radius of 7/8 mm can be created. To avoid Poisson's spo, the symmetry of the problem was broken by circular modulation of the phase leading to a spiral-like structure. The calculated efficiency of the HOE relating the energy within the ring to the total energy was 91%. CONCLUSION: With an HOE it is possible to redistribute the energy along the desired focal ring. The HOE design can be adapted to the intensity distribution of the impinging laser beam with its characteristic aperture shape. A circular homogeneous corneal trephination depth is possible, because the energy fluctuation from pulse to pulse does not locally affect the ablation process. A ring focus for the Er:YAG laser has the potential to render superfluous a manual beam control via micromanipulator and to allow a more rapid and more regular corneal trephination along aperture masks.

Characterization of C3dg binding to a recess formed between short consensus repeats 1 and 2 of complement receptor type 2 (CR2; CD21).

To allow for a better characterization of the ligand binding structures of human complement receptor type 2 (CR2; CD21), we have established an IgG1 kappa mouse mAb, FE8, that interferes efficiently with binding of C3dg and EBV to CR2. In contrast to mAb OKB7, the only well-characterized mAb with similar specificity, mAb FE8 blocked binding of soluble C3dg or particles carrying multiple copies of surface-bound C3dg to CR2 or induced complete removal of these ligands from the receptor. In vitro EBV infection of B lymphocytes, on the other hand, was abrogated by mAbs FE8 and OKB7 with similar dose-response characteristics. As FE8 was shown to recognize a discontinuous epitope, a series of overlapping peptides derived from SCR1 and -2 and immobilized on cellulose was screened with FE8. The results suggest that up to five discontinuous sequences contributed to the epitope. The sequence 63-EYFNKYS-69, located between the two SCR units, reacted most intensively. Two other sequences, 16-YYSTPI-21 and 105-NGNKSVWCQANN-116, are located between Cys1 and Cys2 of SCR1 and around Cys3 of SCR2, respectively. Based on the solution structure for two factor H SCRs, a three-dimensional model of SCR1 and -2 was generated. The FE8 binding peptide sequences were located in relative proximity to each other, bounding the recess formed between SCR1 and -2. This potential of mAb FE8 is currently unique and may be exploited for interfering with conditions of unwanted recognition of C3dg-coated structures by the immune system.