De novo protein structure determination by heavy-atom soaking in lipidic cubic phase and SIRAS phasing using serial synchrotron crystallography.

During the past few years, serial crystallography methods have undergone continuous development and serial data collection has become well established at high-intensity synchrotron-radiation beamlines and XFEL radiation sources. However, the application of experimental phasing to serial crystallography data has remained a challenging task owing to the inherent inaccuracy of the diffraction data. Here, a particularly gentle method for incorporating heavy atoms into micrometre-sized crystals utilizing lipidic cubic phase (LCP) as a carrier medium is reported. Soaking in LCP prior to data collection offers a new, efficient and gentle approach for preparing heavy-atom-derivative crystals directly before diffraction data collection using serial crystallography methods. This approach supports effective phasing by utilizing a reasonably low number of diffraction patterns. Using synchrotron radiation and exploiting the anomalous scattering signal of mercury for single isomorphous replacement with anomalous scattering (SIRAS) phasing resulted in high-quality electron-density maps that were sufficient for building a complete structural model of proteinase K at 1.9 Šresolution using automatic model-building tools.

Prospective Evaluation of the radiologist's hand dose in CT-guided interventions.

PURPOSE: Assessment of radiologist's hand dose in CT-guided interventions and determination of influencing factors. MATERIALS AND METHODS: The following CT-guided interventions were included: Core biopsy, drainage, periradicular therapy, and celiac plexus neurolysis. The hand dose was measured with an immediately readable dosimeter, the EDD-30 (Unfors, Sweden). The default parameters for CT fluoroscopy were 120 kV, 90 mA and a 4 mm slice thickness. All interventions were performed on a 16-slice CT unit (Aquilion 16 Toshiba, Japan). The tumor size, degree of difficulty (1 - 3), level of experience and device parameters (mAs, dose-length product, scan time) were documented. RESULTS: 138 CT-guided interventions (biopsy n = 99, drainage n = 23, pain therapy n = 16) at different locations (lung n = 41, retroperitoneum n = 53, liver n = 25, spine n = 19) were included. The lesion size was 4 - 240 mm (median: 23 mm). The fluoroscopy time per intervention was 4.6 - 140.2 s (median: 24.2 s). The measured hand dose ranged from 0.001 - 3.02 mSv (median: 0.22 mSv). The median hand dose for lung puncture (n = 41) was slightly higher (median: 0.32 mSv, p = 0.01) compared to that for the liver, retroperitoneum and other. Besides physical influencing factors, the degree of difficulty (p = 0.001) and summed puncture depth (p = 0.004) correlated significantly with the hand dose. CONCLUSION: The median hand dose for different CT-guided interventions was 0.22 mSv. Therefore, the annual hand dose limit would normally only be reached with about 2000 interventions.

[Preparation of critical point dried specimens of mouse embryos with a laser scalpel]. / Präparation critical-point-getrockneter Mausembryonen mit einem Laserskalpell.

We investigated the use of a laser scalpel for tissue sparing demonstration of the deep structures of critical point dried specimens. A Nd:YAG-Laser emitted pico-second pulses (wavelength 1064 nm, pulse width 30 ps) at pulse energies varying from 1 microJ to 6 mJ was used. Differences in the effects on sputtered and unsputtered specimens were found. We separate the floor of the mouth and pharyngeal fornix in mouse embryos (10. developmental day). It was concluded the laser scalpel is superior to conventional mechanical dissecting methods when applied to small dried specimens. The advantages and disadvantages of the laser scalpel are discussed.

Mechanisms of intraocular photodisruption with picosecond and nanosecond laser pulses.

Nd:YAG laser photodisruption with nanosecond (ns) pulses is an established method for intraocular surgery. In order to assess whether an increased precision can be achieved by the use of picosecond (ps) pulses, the plasma size, the shock wave characteristics, and the cavitation bubble expansion after optical breakdown with ps- and ns-laser pulses were investigated by time-resolved photography and acoustic measurements. Nd:YAG laser pulses with a duration of 30 ps and 6 ns, respectively, were focused into a water-filled glass cuvette. Frequency doubled light from the same laser pulses was optically delayed between 2 ns and 136 ns and used as illumination light source for photography. Since the individual events were well reproducible, the shock wave and bubble wall position could be determined as a function of time. From the slope of these r(t) curves, the shock wave and bubble wall velocities were determined, and the shock wave pressure was calculated from the shock velocity. The plasma size at various laser pulse energies was measured from photographs of the plasma radiation. The breakdown thresholds at 30 ps and 6 ns pulse duration were found to be 15 microJ and 200 microJ, respectively. At threshold, ps-plasmas are shorter than ns-plasmas, but at the same pulse energy they are always approximately 2.5 times longer. The initial shock pressures were 17 kbar after ps-pulses with an energy of 50 microJ, and 21 kbar after 1 mJ ns-pulses. The pressure amplitude decayed much faster after the ps-pulses. The maximum expansion velocity of the cavitation bubble was 350 m/s after a 50 microJ ps-pulse, but 1,600 m/s after a 1 mJ ns-pulse. The side effects of intraocular microsurgery associated with shock wave emission and cavitation bubble expansion can be considerably reduced by the use of ps-pulses, and new applications of photodisruption may become possible.

[Plasma formation in Nd:YAG laser surgery]. / Untersuchungen zur Plasmabildung bei der Nd:YAG-Laserchirurgie.

Plasma formation with nanosecond and picosecond Nd:YAG laser pulses was investigated as a function of laser pulse energy to find possibilities for a spatial reduction of the laser effects. The threshold for plasma formation (optical breakdown) in distilled water is 200 microJ for ns pulses and 15 microJ for ps pulses. At a pulse energy of 1 mJ, the plasma length is 80 microns for ns pulses and 250 microns for ps pulses. For the same pulse energy, ps plasmas are on average three times larger than ns plasmas. The plasma length is approximately proportional to the cube root of the laser pulse energy for both ns and ps laser pulses. The decrease in the breakdown threshold achieved by using ps pulses renders photodisruption with a smaller pulse energy (in the microJ range) possible, and therefore a reduction in the side effects. The results are compared with the "moving breakdown" model.