Bridging fluorescence microscopy and electron microscopy

Development of new fluorescent probes and fluorescence microscopes has led to new ways to study cell biology. With the emergence of specialized microscopy units at most universities and research centers, the use of these techniques is well within reach for a broad research community. A major breakthrough in fluorescence microscopy in biology is the ability to follow specific targets on or in living cells, revealing dynamic localization and/or function of target molecules. One of the inherent limitations of fluorescence microscopy is the resolution. Several efforts are undertaken to overcome this limit. The traditional and most well-known way to achieve higher resolution imaging is by electron microscopy. Moreover, electron microscopy reveals organelles, membranes, macromolecules, and thus aids in the understanding of cellular complexity and localization of molecules of interest in relation to other structures. With the new probe development, a solid bridge between fluorescence microscopy and electron microscopy is being built, even leading to correlative imaging. This connection provides several benefits, both scientifically as well as practically. Here, I summarize recent developments in bridging microscopy

DNA protection by histone-like protein HU from the hyperthermophilic eubacterium Thermotoga maritima

In mesophilic prokaryotes, the DNA-binding protein HU participates in nucleoid organization as well as in regulation of DNA-dependent processes. Little is known about nucleoid organization in thermophilic eubacteria. We show here that HU from the hyperthermophilic eubacterium Thermotoga maritima HU bends DNA and constrains negative DNA supercoils in the presence of topoisomerase I. However, while binding to a single site occludes ∼35 bp, association of T. maritima HU with DNA of sufficient length to accommodate multiple protomers results in an apparent shorter occluded site size. Such complexes consist of ordered arrays of protomers, as revealed by the periodicity of DNase I cleavage. Association of TmHU with plasmid DNA yields a complex that is remarkably resistant to DNase I-mediated degradation. TmHU is the only member of this protein family capable of occluding a 35 bp nonspecific site in duplex DNA; we propose that this property allows TmHU to form exceedingly stable associations in which DNA flanking the kinks is sandwiched between adjacent proteins. We suggest that T. maritima HU serves an architectural function when associating with a single 35 bp site, but generates a very stable and compact aggregate at higher protein concentrations that organizes and protects the genomic DNA

Spatially Resolving Spin-split Edge States of Chiral Graphene Nanoribbons

A central question in the field of graphene-related research is how graphene behaves when it is patterned at the nanometer scale with different edge geometries. Perhaps the most fundamental shape relevant to this question is the graphene nanoribbon (GNR), a narrow strip of graphene that can have different chirality depending on the angle at which it is cut. Such GNRs have been predicted to exhibit a wide range of behaviour (depending on their chirality and width) that includes tunable energy gaps and the presence of unique one-dimensional (1D) edge states with unusual magnetic structure. Most GNRs explored experimentally up to now have been characterized via electrical conductivity, leaving the critical relationship between electronic structure and local atomic geometry unclear (especially at edges). Here we present a sub-nm-resolved scanning tunnelling microscopy (STM) and spectroscopy (STS) study of GNRs that allows us to examine how GNR electronic structure depends on the chirality of atomically well-defined GNR edges. The GNRs used here were chemically synthesized via carbon nanotube (CNT) unzipping methods that allow flexible variation of GNR width, length, chirality, and substrate. Our STS measurements reveal the presence of 1D GNR edge states whose spatial characteristics closely match theoretical expectations for GNR's of similar width and chirality. We observe width-dependent splitting in the GNR edge state energy bands, providing compelling evidence of their magnetic nature. These results confirm the novel electronic behaviour predicted for GNRs with atomically clean edges, and thus open the door to a whole new area of applications exploiting the unique magnetoelectronic properties of chiral GNRs

Field-induced p-n transition in yttria-stabilized zirconia

Oxide ion conducting yttria-stabilised zirconia ceramics show the onset of electronic conduction under a small bias voltage. Compositions with a high yttria content undergo a transition from p-type to n-type behavior at voltages in the range 2.4 to 10 V, which also depends on oxygen partial pressure. Surface reactions have a direct influence on bulk electronic conductivities, with possible implications for voltage-induced flash phenomena and resistive switching

Ferromagnetism in graphene nanoribbons: split versus oxidative unzipped ribbons

Two types of graphene nanoribbons: (a) potassium-split graphene nanoribbons (GNRs), and (b) oxidative unzipped and chemically converted graphene nanoribbons (CCGNRs) were investigated for their magnetic properties using the combination of static magnetization and electron spin resonance measurements. The two types of ribbons possess remarkably different magnetic properties. While the low temperature ferromagnet-like feature is observed in both types of ribbons, such room temperature feature persists only in potassium-split ribbons. The GNRs show negative exchange bias, but the CCGNRs exhibit a 'positive exchange bias'. Electron spin resonance measurements infer that the carbon related defects may responsible for the observed magnetic behaviour in both types of ribbons. Furthermore, proton hyperfine coupling strength has been obtained from hyperfine sublevel correlation experiments performed on the GNRs. Electron spin resonance provides no indications for the presence of potassium (cluster) related signals, emphasizing the intrinsic magnetic nature of the ribbons. Our combined experimental results may infer the coexistence of ferromagnetic clusters with anti-ferromagnetic regions leading to disordered magnetic phase. We discuss the origin of the observed contrast in the magnetic behaviours of these two types of ribbons

Association between erythrocyte Na+K+-ATPase activity and some blood lipids in type 1 diabetic patients from Lagos, Nigeria

<p>Abstract</p> <p>Background</p> <p>Altered levels of erythrocyte Na⁺K⁺-ATPase, atherogenic and anti-atherogenic lipid metabolites have been implicated in diabetic complications but their pattern of interactions remains poorly understood.</p> <p>This study evaluated this relationship in Nigerian patients with Type 1 diabetes mellitus.</p> <p>Methods</p> <p>A total of 34 consented Type 1 diabetic patients and age -matched 27 non-diabetic controls were enrolled. Fasting plasma levels of total cholesterol, triglycerides and HDL-cholesterol were determined spectrophotometrically and LDL-cholesterol estimated using Friedewald formula. Total protein content and Na+K+-ATPase activity were also determined spectrophotometrically from ghost erythrocyte membrane prepared by osmotic lysis.</p> <p>Results</p> <p>Results indicate significant (P < 0.05) reduction in Na⁺K⁺-ATPase activity in the Type 1 diabetic patients (0.38 ± 0.08 vs. 0.59 ± 0.07 uM Pi/mgprotein/h) compared to the control but with greater reduction in the diabetic subgroup with poor glycemic control (n = 20) and in whom cases of hypercholesterolemia (8.8%), hypertriglyceridemia (2.9%) and elevated LDL-cholesterol (5.9% each) were found. Correlation analyses further revealed significant (P < 0.05) inverse correlations [r = -(0.708-0.797] between all the atherogenic lipid metabolites measured and Na⁺K⁺-ATPase in this subgroup contrary to group with good glycemic control or non-diabetic subjects in which significant (P < 0.05) Na⁺K⁺-ATPase and HDL-C association were found (r = 0.427 - 0.489). The Na⁺K⁺-ATPase from the diabetic patients also exhibited increased sensitivity to digoxin and alterations in kinetic constants Vmax and Km determined by glycemic status of the patients.</p> <p>Conclusion</p> <p>It can be concluded that poor glycemic control evokes greater reduction in erythrocyte Na⁺K⁺-ATPase activity and promote enzyme-blood atherogenic lipid relationships in Type 1 diabetic Nigerian patients.</p

Evidence of Activity-Specific, Radial Organization of Mitotic Chromosomes in Drosophila

A fluorescently labeled protein specifically binding to genes was reproducibly found at the periphery of condensed mitotic fruit fly chromosomes, illustrating preservation of a radial structure between consecutive divisions