Constructing a cost-efficient, high-throughput and high-quality single-molecule localization microscope for super-resolution imaging.

Single-molecule localization microscopy (SMLM) leverages the power of modern optics to unleash ultra-precise structural nanoscopy of complex biological machines in their native environments as well as ultra-sensitive and high-throughput medical diagnostics with the sensitivity of a single molecule. To achieve this remarkable speed and resolution, SMLM setups are either built by research laboratories with strong expertise in optical engineering or commercially sold at a hefty price tag. The inaccessibility of SMLM to life scientists for technical or financial reasons is detrimental to the progress of biological and biomedical discoveries reliant on super-resolution imaging. In this work, we present the NanoPro, an economic, high-throughput, high-quality and easy-to-assemble SMLM for super-resolution imaging. We show that our instrument performs similarly to the most expensive, best-in-class commercial microscopes and rivals existing open-source microscopes at a lower price and construction complexity. To facilitate its wide adoption, we compiled a step-by-step protocol, accompanied by extensive illustrations, to aid inexperienced researchers in constructing the NanoPro as well as assessing its performance by imaging ground-truth samples as small as 20 nm. The detailed visual instructions make it possible for students with little expertise in microscopy engineering to construct, validate and use the NanoPro in <1 week, provided that all components are available.

Quantum mechanics-free subsystem with mechanical oscillators.

Quantum mechanics sets a limit for the precision of continuous measurement of the position of an oscillator. We show how it is possible to measure an oscillator without quantum back-action of the measurement by constructing one effective oscillator from two physical oscillators. We realize such a quantum mechanics-free subsystem using two micromechanical oscillators, and show the measurements of two collective quadratures while evading the quantum back-action by 8 decibels on both of them, obtaining a total noise within a factor of 2 of the full quantum limit. This facilitates the detection of weak forces and the generation and measurement of nonclassical motional states of the oscillators. Moreover, we directly verify the quantum entanglement of the two oscillators by measuring the Duan quantity 1.4 decibels below the separability bound.

Femtosecond fiber Bragg grating fabrication with adaptive optics aberration compensation.

We present fiber Bragg gratings (FBGs) fabricated using adaptive optics aberration compensation for the first time to the best of our knowledge. The FBGs are fabricated with a femtosecond laser by the point-by-point method using an air-based objective lens, removing the requirement for immersion oil or ferrules. We demonstrate a general phase correction strategy that can be used for accurate fabrication at any point in the fiber cross-section. We also demonstrate a beam-shaping approach that nullifies the aberration when focused inside a central fiber core. Both strategies give results which are in excellent agreement with coupled-mode theory. An extremely low wavelength polarization sensitivity of 4 pm is reported.

Decarboxylative-coupling of allyl acetate catalyzed by group 10 organometallics, [(phen)M(CH3)]+.

Gas-phase carbon-carbon bond forming reactions, catalyzed by group 10 metal acetate cations [(phen)M(O2CCH3)](+) (where M = Ni, Pd or Pt) formed via electrospray ionization of metal acetate complexes [(phen)M(O2CCH3)2], were examined using an ion trap mass spectrometer and density functional theory (DFT) calculations. In step 1 of the catalytic cycle, collision induced dissociation (CID) of [(phen)M(O2CCH3)](+) yields the organometallic complex, [(phen)M(CH3)](+), via decarboxylation. [(phen)M(CH3)](+) reacts with allyl acetate via three competing reactions, with reactivity orders (% reaction efficiencies) established via kinetic modeling. In step 2a, allylic alkylation occurs to give 1-butene and reform metal acetate, [(phen)M(O2CCH3)](+), with Ni (36%) > Pd (28%) > Pt (2%). Adduct formation, [(phen)M(C6H11O2)](+), occurs with Pt (24%) > Pd (21%) > Ni(11%). The major losses upon CID on the adduct, [(phen)M(C6H11O2)](+), are 1-butene for M = Ni and Pd and methane for Pt. Loss of methane only occurs for Pt (10%) to give [(phen)Pt(C5H7O2)](+). The sequences of steps 1 and 2a close a catalytic cycle for decarboxylative carbon-carbon bond coupling. DFT calculations suggest that carbon-carbon bond formation occurs via alkene insertion as the initial step for all three metals, without involving higher oxidation states for the metal centers.

Web-based post-traumatic stress disorder education for military family members.

OBJECTIVE: Since post-traumatic stress disorder (PTSD) is common after military deployment and affects both military service members and their families, we sought to both improve PTSD-related knowledge of military family members and to foster actions to help service members with their symptoms. METHODS: Focus groups were conducted with military family members and their feedback was incorporated into an educational Web site to improve family members' knowledge of PTSD. We pilot-tested the site and a 25-item questionnaire, then used it to assess the knowledge of 497 family members before and after their use of the Web site. RESULTS: Use of this educational Web site improved military family members' PTSD-related knowledge on a 25-item test, with an increase from a mean 13.9 correct responses beforehand to 18.7 after Web site use (p < 0.001; effect size 1.2). In addition, 217 family members returned to the site > or =10 days after their initial visit; 57% had taken actions such as discussing the service member's symptoms with them or persuading them to get medical attention, and 82 to 95% of them believed their actions to be beneficial. CONCLUSION: A Web-based intervention can both improve PTSD-related knowledge and foster behavioral changes in military family members.

Nitrogen adduction by three coordinate group 10 organometallic cations: platinum is favoured over nickel and palladium.

Previous studies have shown that highly reactive product ions formed by collision-induced dissociation (CID) of precursor ions generated via electrospray can readily react with residual solvent or drying gases, especially in ion trap mass spectrometers. Here we report on the rapid addition of nitrogen to the coordinatively unsaturated organoplatinum cation, [(phen)Pt(CH(3))](+) (phen=1,10-phenanthroline) formed via decarboxylation of the acetate complex [(phen)Pt(O(2) CCH(3))](+). This contrasts with the related coordinatively unsaturated group 10 cations: addition of nitrogen to [(phen)Pd(CH(3))](+) occurs at longer reaction times, whereas addition of nitrogen to [(phen)Ni(CH(3))](+) is virtually non-existent. To better understand these reactions, density functional theory (DFT) calculations were carried out at the B3LYP/SDD6-31+G(d) level of theory to determine the N(2)-binding energies of [(phen)M(CH(3))](+). [(phen)Pt(CH(3))](+) has a higher binding energy to N(2) (1.06 eV) than either [(phen)Ni(CH(3))](+) (0.61 eV) or [(phen)Pd(CH(3))](+) (0.66 eV), consistent with the experimental ease of addition of nitrogen to the coordinatively unsaturated organometallic complexes, [(phen)M(CH(3))](+). Finally, [(phen)M(CH(3))](+) are reactive to other background gases, forming [(phen)M(O(2))](.+) (for M=Ni) in reactions with oxygen and undergoing water addition (for M=Ni, Pd and Pt) and water addition/CH(4) elimination reactions to yield [(phen)M(OH)](+) (for M=Ni and Pt).

Effect of freezing parameters (freeze cycle and thaw process) on tissue destruction following renal cryoablation.

BACKGROUND AND PURPOSE: Renal cryoablation is a successful nephron-sparing treatment alternative for selected patients with small renal tumors. The purpose of this study was to compare the effects of the number of freeze cycles (one v two) and the thaw process (active v passive) on renal tissue following cryodestruction. MATERIALS AND METHODS: Sixteen female mongrel dogs (19.9 +/- 2.1 kg) were randomly divided into four groups and underwent transabdominal laparoscopic access by standard techniques. Tissue freezing was performed using argon gas following interstitial cryoprobe (3 mm) placement into the upper and lower poles of the left kidney. Single active (SA), single passive (SP) double active (DA) or double passive (DP) 15-minute treatment cycle(s) were carried out via the CRYOcare Cryosurgical Unit (Endocare, Irving, CA) on eight kidneys each. An active thaw process with helium gas or a passive thaw process was initiated after each freeze period. The cryoprobe was removed when the temperature reached 0 degrees C. Four weeks following cryosurgery, animals were sacrificed, and the renal tissue was evaluated grossly and histologically. RESULTS: Interstitial cryoprobe temperatures decreased from 31.3 degrees C +/- 1.4 degrees C to -142 degrees C +/- 1.0 degrees C following the 15-minute freeze cycle. The temperature reached 0 degrees C significantly faster following active thaw than with the passive process (2.13 +/- 0.24 min/freeze cycle and 15.18 +/- 2.97 min/freeze cycle, respectively; P < 0.0001). Grossly, each lesion consisted of a central area of necrosis surrounded by a rim of white tissue. On microscopic examination, each lesion consisted of a central area of liquefaction necrosis (LN) surrounded by various degrees of fibrosis and granulation tissue admixed with residual parenchyma. The size of the LN was significantly different in tissues subjected to double and single freeze cycles when compared across both thaw processes (active and passive). There was no significant difference in the overall lesion volume following DA, DP, SA, or SP. CONCLUSIONS: Renal cryodestruction via laparoscopic access achieves complete tissue ablation without complications. The double freeze cycle produced significantly larger areas of LN than the single freeze regardless of the thaw process. The type of thaw process did not affect the amount of tissue damage. Utilizing a double 15-minute freeze cycle with the faster active thaw process will effectively cryoablate renal tissue as well as significantly reduce overall operative time.