Microrheology imaging of fiber suspensions – a case study for lyophilized collagen I in HCl solutions

In fiber suspensions with low optical contrast, the in situ characterization of structural properties with conventional microscopy methods fails. However, overlaying subsequent images of multiple particle tracking (MPT) videos including short trajectories usually discarded in MPT analysis allowed for direct visualization of individual fibers and the network structure of lyophilized collagen I (Coll) distributed in hydrochloric acid solutions. MPT yielded a broad distribution of mean square displacements (MSDs). Freely diffusing tracer particles yielded viscosities indicating that, irrespective of concentration, a constant amount of Coll is dissolved in the aqueous phase. Particles found elastically trapped within fibrous Coll structures exhibited a broad range of time-independent MSDs and we propose a structure comprising multiple fiber bundles with dense regions inaccessible to tracers and elastic regions of different stiffness in between. Bulky aggregates inaccessible to the 0.2 μm tracers exist even at low Coll concentrations, a network of slender fibers evolves above the sol–gel transition and these fibers densify with increasing Coll concentration. This novel MPT-based imaging technique possesses great potential to characterize the fiber distribution in and structural properties of a broad range of biological and technical suspensions showing low contrast when imaged with conventional techniques. Thus, MPT imaging and microrheology will help to better understand the effect of fiber distribution and network structure on the viscoelastic properties of complex suspensions

Radical Ions in the Pentalene Series. Part III. Three Paramagnetic Redox Stages of a Dicyclopenta[a,e]pentalene

Five redox stages have been observed for the recently synthesized 1,3,5,7-tetra(tert-butyl) derivative 1 of the dicyclopenta[a,e]pentalene, a novel non-alternant hydrocarbon: the radical cation 1 , the neutral compound 1, the radical anion Is, the dianion 1²-, and the radical trianion 1³. Information about the electronic structure of the three paramagnetic stages, 1, 1⁻, and 1³⁻ is provided by the use of ESR, ENDOR, and TRIPLE resonance spectroscopy. The unpaired electron in the trianion resides mainly on the ‘inner’ butadiene-π-system, whereas in the cation and the anion, it is largely localized on the two ‘outer’ five-membered rings

Synthesis and Properties of the First [4.4]Ferrocenophane-1,3,15,17-tetrayne

Building butadiyne bridges: The kinetically stabilized 1,1′-diethynylferrocene, readily prepared from the pentafulvenoid allene 1, can be transformed into the [4.4]ferrocenophane 2 by oxidative coupling. This compound exhibits a remarkably symmetrical structure in which the electrons are delocalized through the butadiyne bridges; in the crystalline state 2 exists in a helical-chiral conformation

Synthese und Eigenschaften des ersten [4.4]Ferrocenophan-1,3,15,17-tetrains

Das aus dem pentafulvenoiden Allen 1 leicht zugängliche, kinetisch stabilisierte 1,1′-Diethinylferrocen lässt sich durch oxidative Kupplung in das [4.4]Ferrocenophan 2 überführen. Diese Verbindung hat eine bemerkenswert symmetrische Struktur, die Elektronen sind über die Butadiinbrücken hinweg delokalisiert, und im Kristall liegt 2 in einer helical-chiralen Konformation vor

A Novel Optical Tissue Clearing Protocol for Mouse Skeletal Muscle to Visualize Endplates in Their Tissue Context

Neuromuscular junctions (NMJs) mediate skeletal muscle contractions and play an important role in several neuromuscular disorders when their morphology and function are compromised. However, due to their small size and sparse distribution throughout the comparatively large, inherently opaque muscle tissue the analysis of NMJ morphology has been limited to teased fiber preparations, longitudinal muscle sections, and flat muscles. Consequently, whole mount analyses of NMJ morphology, numbers, their distribution, and assignment to a given muscle fiber have also been impossible to determine in muscle types that are frequently used in experimental paradigms. This impossibility is exacerbated by the lack of optical tissue clearing techniques that are compatible with clear and persistent NMJ stains. Here, we present MYOCLEAR, a novel and highly reproducible muscle tissue clearing protocol. Based on hydrogel-based tissue clearing methods, this protocol permits the labeling and detection of all NMJs in adult hindleg extensor digitorum longus muscles from wildtype and diseased mice. The method is also applicable to adult mouse diaphragm muscles and can be used for different staining agents, including toxins, lectins, antibodies, and nuclear dyes. It will be useful in understanding the distribution, morphological features, and muscle tissue context of NMJs in hindleg muscle whole mounts for biomedical and basic research

A Novel Optical Tissue Clearing Protocol for Mouse Skeletal Muscle to Visualize Endplates in Their Tissue Context

Neuromuscular junctions (NMJs) mediate skeletal muscle contractions and play an important role in several neuromuscular disorders when their morphology and function are compromised. However, due to their small size and sparse distribution throughout the comparatively large, inherently opaque muscle tissue the analysis of NMJ morphology has been limited to teased fiber preparations, longitudinal muscle sections, and flat muscles. Consequently, whole mount analyses of NMJ morphology, numbers, their distribution, and assignment to a given muscle fiber have also been impossible to determine in muscle types that are frequently used in experimental paradigms. This impossibility is exacerbated by the lack of optical tissue clearing techniques that are compatible with clear and persistent NMJ stains. Here, we present MYOCLEAR, a novel and highly reproducible muscle tissue clearing protocol. Based on hydrogel-based tissue clearing methods, this protocol permits the labeling and detection of all NMJs in adult hindleg extensor digitorum longus muscles from wildtype and diseased mice. The method is also applicable to adult mouse diaphragm muscles and can be used for different staining agents, including toxins, lectins, antibodies, and nuclear dyes. It will be useful in understanding the distribution, morphological features, and muscle tissue context of NMJs in hindleg muscle whole mounts for biomedical and basic research.</p