Soft x-ray laser beamline for surface processing and damage studies.

We have developed a soft x-ray laser (SXRL) beamline equipped with an intensity monitor dedicated to ablation study such as surface processing and damage formation. The SXRL beam having a wavelength of 13.9 nm, pulse width of 7 ps, and pulse energy of around 200 nJ is generated from Ag plasma mediums using an oscillator-amplifier configuration. The SXRL beam is focused onto the sample surface by the Mo/Si multilayer coated spherical mirror. To get the correct irradiation energy/fluence, an intensity monitor composed of a Mo/Si multilayer beam splitter and an x-ray charge-coupled device camera has been installed in the beamline. The Mo/Si multilayer beam splitter has a large polarization dependence in the reflectivity around the incident angle of 45°. However, by evaluating the relationship between reflectivity and transmittance of the beam splitter appropriately, the irradiation energy onto the sample surface can be derived from the energy acquired by the intensity monitor. This SXRL beamline is available to not only the ablation phenomena but also the performance evaluation of soft x-ray optics and resists.

Single-cell temperature mapping with fluorescent thermometer nanosheets.

Recent studies using intracellular thermometers have shown that the temperature inside cultured single cells varies heterogeneously on the order of 1°C. However, the reliability of intracellular thermometry has been challenged both experimentally and theoretically because it is, in principle, exceedingly difficult to exclude the effects of nonthermal factors on the thermometers. To accurately measure cellular temperatures from outside of cells, we developed novel thermometry with fluorescent thermometer nanosheets, allowing for noninvasive global temperature mapping of cultured single cells. Various types of cells, i.e., HeLa/HEK293 cells, brown adipocytes, cardiomyocytes, and neurons, were cultured on nanosheets containing the temperature-sensitive fluorescent dye europium (III) thenoyltrifluoroacetonate trihydrate. First, we found that the difference in temperature on the nanosheet between nonexcitable HeLa/HEK293 cells and the culture medium was less than 0.2°C. The expression of mutated type 1 ryanodine receptors (R164C or Y523S) in HEK293 cells that cause Ca2+ leak from the endoplasmic reticulum did not change the cellular temperature greater than 0.1°C. Yet intracellular thermometry detected an increase in temperature of greater than ∼2°C at the endoplasmic reticulum in HeLa cells upon ionomycin-induced intracellular Ca2+ burst; global cellular temperature remained nearly constant within ±0.2°C. When rat neonatal cardiomyocytes or brown adipocytes were stimulated by a mitochondrial uncoupling reagent, the temperature was nearly unchanged within ±0.1°C. In cardiomyocytes, the temperature was stable within ±0.01°C during contractions when electrically stimulated at 2 Hz. Similarly, when rat hippocampal neurons were electrically stimulated at 0.25 Hz, the temperature was stable within ±0.03°C. The present findings with nonexcitable and excitable cells demonstrate that heat produced upon activation in single cells does not uniformly increase cellular temperature on a global basis, but merely forms a local temperature gradient on the order of ∼1°C just proximal to a heat source, such as the endoplasmic/sarcoplasmic reticulum ATPase.