Addressing the Requirements of High‐Sensitivity Single‐Molecule Imaging of Low‐Copy‐Number Proteins in Bacteria

Single‐molecule fluorescence super‐resolution imaging and tracking provide nanometer‐scale information about subcellular protein positions and dynamics. These single‐molecule imaging experiments can be very powerful, but they are best suited to high‐copy number proteins where many measurements can be made sequentially in each cell. We describe artifacts associated with the challenge of imaging a protein expressed in only a few copies per cell. We image live Bacillus subtilis in a fluorescence microscope, and demonstrate that under standard single‐molecule imaging conditions, unlabeled B. subtilis cells display punctate red fluorescent spots indistinguishable from the few PAmCherry fluorescent protein single molecules under investigation. All Bacillus species investigated were strongly affected by this artifact, whereas we did not find a significant number of these background sources in two other species we investigated, Enterococcus faecalis and Escherichia coli. With single‐molecule resolution, we characterize the number, spatial distribution, and intensities of these impurity spots.Bright spots: A single‐molecule‐like fluorescent background signal is reported in Bacillus subtilis cells, and the density and fluorescence intensity of these spots are quantified in several Bacillus species and other Gram‐negative and Gram‐positive organisms.Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/144710/1/cphc201600035_am.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/144710/2/cphc201600035.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/144710/3/cphc201600035-sup-0001-misc_information.pd

Magnetic field dependence of superconducting energy gaps in YNi2B2C: Evidence of multiband superconductivity

We present results of in field directional point contact spectroscopy (DPCS) study in the quaternary borocarbide superconductor YNi2B2C, which is characterized by a highly anisotropic superconducting gap function. For I||a, the superconducting energy gap (D), decreases linearly with magnetic field and vanishes around 3.25T which is well below the upper critical field (Hc2~6T) measured at the same temperature (2.2K). For I||c, on the other hand, D decreases weakly with magnetic field but the broadening parameter (G) increases rapidly with magnetic field with the absence of any resolvable feature above 3.5T. From an analysis of the field variation of energy gaps and the zero bias density of states we show that the unconventional gap function observed in this material could originate from multiband superconductivity.Comment: 19 pages including figures (final version

Human monoclonal islet specific autoantibodies share features of islet cell and 64 kDa antibodies

The first human monoclonal islet cell antibodies of the IgG class (MICA 1-6) obtained from an individual with Type 1 (insulin-dependent) diabetes mellitus were cytoplasmic islet cell antibodies selected by the indirect immunofluorescence test on pancreas sections. Surprisingly, they all recognized the 64 kDa autoantigen glutamate decarboxylase. In this study we investigated which typical features of cytoplasmic islet cell antibodies are represented by these monoclonals. We show by double immunofluorescence testing that MICA 1-6 stain pancreatic beta cells which is in agreement with the beta-cell specific expression of glutamate decarboxylase. In contrast an islet-reactive IgM monoclonal antibody obtained from a pre-diabetic individual stained all islet cells but lacked the tissue specificity of MICA 1-6 and must therefore be considered as a polyreactive IgM-antibody. We further demonstrate that MICA 1-6 revealed typical features of epitope sensitivity to biochemical treatment of the target tissue which has been demonstrated for islet cell antibodies, and which has been used to argue for a lipid rather than a protein nature of target antigens. Our results provide direct evidence that the epitopes recognized by the MICA are destroyed by methanol/chloroform treatment but reveal a high stability to Pronase digestion compared to proinsulin epitopes. Conformational protein epitopes in glutamate decarboxylase therefore show a sensitivity to biochemical treatment of sections such as ganglioside epitopes. MICA 1-6 share typical features of islet cell and 64 kDa antibodies and reveal that glutamate decarboxylase-reactive islet cell antibodies represent a subgroup of islet cell antibodies present in islet cell antibody-positive sera

A Novel Dielectric Anomaly in Cuprates and Nickelates: Signature of an Electronic Glassy State

The low-frequency dielectric response of hole-doped insulators La_{2}Cu_{1-x}Li_{x}O_{4} and La_{2-x}Sr_{x}NiO_{4} shows a large dielectric constant \epsilon ^{'} at high temperature and a step-like drop by a factor of 100 at a material-dependent low temperature T_{f}. T_{f} increases with frequency and the dielectric response shows universal scaling in a Cole-Cole plot, suggesting that a charge glass state is realized both in the cuprates and in the nickelates.Comment: 5 pages, 4 figure

Hidden Magnetism and Quantum Criticality in the Heavy Fermion Superconductor CeRhIn5

With understood exceptions, conventional superconductivity does not coexist with long-range magnetic order[1]. In contrast, unconventional superconductivity develops near a boundary separating magnetically ordered and magnetically disordered phases[2,3]. A maximum in the superconducting transition temperature Tc develops where this boundary extrapolates to T=0 K, suggesting that fluctuations associated with this magnetic quantum-critical point are essential for unconventional superconductivity[4,5]. Invariably though, unconventional superconductivity hides the magnetic boundary when T < Tc, preventing proof of a magnetic quantum-critical point[5]. Here we report specific heat measurements of the pressure-tuned unconventional superconductor CeRhIn5 in which we find a line of quantum-phase transitions induced inside the superconducting state by an applied magnetic field. This quantum-critical line separates a phase of coexisting antiferromagnetism and superconductivity from a purely unconventional superconducting phase and terminates at a quantum tetracritical point where the magnetic field completely suppresses superconductivity. The T->0 K magnetic field-pressure phase diagram of CeRhIn5 is well described with a theoretical model[6,7] developed to explain field-induced magnetism in the high-Tc cuprates but in which a clear delineation of quantum-phase boundaries has not been possible. These experiments establish a common relationship among hidden magnetism, quantum criticality and unconventional superconductivity in cuprate and heavy-electron systems, such as CeRhIn5.Comment: journal reference adde

Fluctuation Study of the Specific Heat of MgB2

The specific heat of polycrystalline Mg$^{11}$B$_{2}$ has been measured with high resolution ac calorimetry from 5 to 45 K at constant magnetic fields. The excess specific heat above T$_{c}$ is discussed in terms of Gaussian fluctuations and suggests that Mg$^{11}$B$_{2}$ is a bulk superconductor with Ginzburg-Landau coherence length $\xi_{0}=26$ \AA . The transition-width broadening in field is treated in terms of lowest-Landau-level (LLL) fluctuations. That analysis requires that $\xi_{0}=20$ \AA . The underestimate of the coherence length in field, along with deviations from 3D LLL predictions, suggest that there is an influence from the anisotropy of B$_{c2}$ between the c-axis and the a-b plane.Comment: Phys. Rev. B 66, 134515 (2002

Specific Heat Study of the Magnetic Superconductor HoNi2B2C

The complex magnetic transitions and superconductivity of HoNi2B2C were studied via the dependence of the heat capacity on temperature and in-plane field angle. We provide an extended, comprehensive magnetic phase diagram for B // [100] and B // [110] based on the thermodynamic measurements. Three magnetic transitions and the superconducting transition were clearly observed. The 5.2 K transition (T_{N}) shows a hysteresis with temperature, indicating the first order nature of the transition at B=0 T. The 6 K transition (T_{M}), namely the onset of the long-range ordering, displays a dramatic in-plane anisotropy: T_{M} increases with increasing magnetic field for B // [100] while it decreases with increasing field for B // [110]. The anomalous anisotropy in T_{M} indicates that the transition is related to the a-axis spiral structure. The 5.5 K transition (T^{*}) shows similar behavior to the 5.2 K transition, i.e., a small in-plane anisotropy and scaling with Ising model. This last transition is ascribed to the change from a^{*} dominant phase to c^{*} dominant phase.Comment: 9 pages, 11 figure