摘要： Reversibly switchable fluorescent proteins (RSFPs) can be effectively used for super-resolution optical fluctuation imaging (SOFI) based on the switching and fluctuation of single molecules. Several properties of RSFPs strongly influence the quality of SOFI images. These properties include (i) the averaged fluorescence intensity in the fluctuation state, (ii) the on/off contrast ratio, (iii) the photostability, and (iv) the oligomerization tendency. The first three properties determine the fluctuation range of the imaged pixels and the SOFI signal, which are of essential importance to the spatial resolution, and the last may lead to artificial aggregation of target proteins. The RSFPs that are currently used for SOFI are low in averaged fluorescence intensity in the fluctuation state, photostability, and on/off contrast ratio, thereby limiting the range of application of SOFI in biological super-resolution imaging. In this study, we developed a novel monomeric green RSFP termed Skylan-S, which features very high photostability, contrast ratio, and averaged fluorescence intensity in the fluctuation state. Taking advantage of the excellent optical properties of Skylan-S, a 4-fold improvement in the fluctuation range of the imaged pixels and higher SOFI resolution can be obtained compared with Dronpa. Furthermore, super-resolution imaging of the actin or tubulin structures and clathrin-coated pits (CCPs) in living U2OS cells labeled with Skylan-S was demonstrated using the SOFI technique. Overall, Skylan-S developed with outstanding photochemical properties is promising for long-time SOH imaging with high spatial-temporal resolution.
摘要： Co-phase and co-focus detection is one of the key technologies for large-aperture segmented mirror telescopes. In this paper, a new edge sensor based on fringes of equal thickness is developed, which can detect each segment's relative piston, tilt, and tip errors from the interferograms. Based on the co-focus demand for many ground-based seeing limited segmented mirror telescopes, an edge sensor prototype based on such a principle is built and applied in the indoor segmented mirror experiment system in the lab. According to the co-focus requirement of the Large Sky Area Multi-Object Fiber Spectroscopic Telescope, many simulations and experiments are carried out for co-focus error detection of the segmented mirror system. Experiment results show that the co-focus accuracy is better than 0."02 rms, which can meet the co-focus requirements of most large or extremely large segmented mirror astronomical telescopes.
摘要： Super-resolution fluorescence microscopy is distinct among nanoscale imaging tools in its ability to image protein dynamics in living cells. Structured illumination microscopy (SIM) stands out in this regard because of its high speed and low illumination intensities, but typically offers only a twofold resolution gain. We extended the resolution of live-cell SIM through two approaches: ultrahigh numerical aperture SIM at 84-nanometer lateral resolution for more than 100 multicolor frames, and nonlinear SIM with patterned activation at 45- to 62-nanometer resolution for approximately 20 to 40 frames. We applied these approaches to image dynamics near the plasma membrane of spatially resolved assemblies of clathrin and caveolin, Rab5a in early endosomes, and a-actinin, often in relationship to cortical actin. In addition, we examined mitochondria, actin, and the Golgi apparatus dynamics in three dimensions.
摘要： We carry out a systematical study of the spectral lag properties of 50 single-pulsed Gamma-Ray Bursts (GRBs) detected by Fermi/GBM. By dividing the light curves into multiple consecutive energy channels we provide a new measurement of the spectral lag which is independent on energy channel selections. We perform a detailed statistical study of our new measurements. We find two similar power-law energy dependencies of both the pulse arrival time and pulse width. Our new results on the power-law indices would favor the relativistic geometric effects for the origin of spectral lag. However, a complete theoretical framework that can fully account for the diverse energy dependencies of both arrival time and pulse width revealed in this work is still missing. We also study the spectral evolution behaviors of the GRB pulses. We find that the GRB pulse with negligible spectral lag would usually have a shorter pulse duration and would appear to have a ``hardness-intensity tracking'' (HIT) behavior and the GRB pulse with a significant spectral lag would usually have a longer pulse duration and would appear to have a ``hard-to-soft'' (HTS) behavior.
摘要： A novel dipeptidyl peptidase IV inhibitor hit (5, IC50 = 0.86 mu M) was structurally derived from our recently disclosed preclinical candidate 4 by replacing the cyanobenzyl with a butynyl based on pharmacophore hybridization. A hit-to-lead optimization effort was then initiated to improve its potency. Most N-substituted analogs exhibited good in vitro activity, and compound 180 (IC50 = 1.55 nM) was identified to be a potent dipeptidyl peptidase IV inhibitor with a significantly improved pharmacokinetic properties (bioavailablity: 41% vs 82.9%; T-1/2: 2 h vs 4.9 h). (C) 2013 Elsevier Ltd. All rights reserved.