Combining X-ray excited optical luminescence and X-ray absorption spectroscopy for correlative imaging on the nanoscale

X-ray absorption and optical luminescence can both provide valuable but very different information on the chemical and physical properties of materials. Although it is known that the spectral characteristics of many materials are highly heterogeneous on the micro- and/or nanoscale, no methodology has so far been shown to be capable of spatially resolving both full X-ray absorption and X-ray excited optical luminescence (XEOL) spectra on the nanoscale in a correlative manner. For this purpose, the scanning transmission X-ray microscope at the HERMES beamline of the SOLEIL synchrotron was equipped with an optical detection system capable of recording high-resolution XEOL spectra using a 40 nm soft X-ray probe. The functionality of the system was demonstrated by analyzing ZnO powder dispersions — showing simultaneously the X-ray linear dichroism and XEOL behavior of individual submicrometric ZnO crystallites.

Steady-state ferumoxytol-enhanced MRI: early observations in benign abdominal organ masses and clinical implications

Introduction The off-label use of ferumoxytol as a vascular MR imaging agent is growing rapidly. However, the properties of ferumoxytol suggest that it may play an important role in the detection and characterization of abdominal mass lesions. Methods Thirty-six patients with benign abdominal mass lesions who underwent MR angiography with ferumoxytol also had T2-weighted HASTE imaging and fat-suppressed 3D T1-weighted imaging. The T1 and T2 enhancement characteristics of the lesions were analyzed and correlated with other imaging modalities and/or surgical findings and/or clinical follow-up. Results In all patients with benign masses in the liver (n = 22 patients), spleen (n = 6 patients), kidneys (n = 33 patients), adrenal (n = 2 patients) and pancreas (n = 4 patients), based on the enhancement characteristics with ferumoxytol, readers were confident of the benign nature of the lesions and their conclusions were consistent with correlative imaging, tissue sampling and follow-up. One patient with a suspicious enhancing 2F Bosniak renal cyst had renal cell carcinoma confirmed on biopsy. Conclusion Ferumoxytol-enhanced MRI can increase diagnostic confidence for benign abdominal masses and can increase the conspicuity of mass lesions, relative to unenhanced MRI. Graphic Abstract

Imaging of retina cellular and subcellular structures using ptychographic hard X-ray tomography

ABSTRACT Ptychographic hard X-ray computed tomography (PXCT) is a recent method allowing imaging with quantitative electron-density contrast. Here, we imaged, at cryogenic temperature and without sectioning, cellular and subcellular structures of a chemically fixed and stained wild-type mouse retina, including axons and synapses, with complete isotropic 3D information over tens of microns. Comparison with tomograms of degenerative retina from a mouse model of retinitis pigmentosa illustrates the potential of this method for analyzing disease processes like neurodegeneration at sub-200 nm resolution. As a non-destructive imaging method, PXCT is very suitable for correlative imaging. Within the outer plexiform layer containing the photoreceptor synapses, we identified somatic synapses. We used a small region inside the X-ray-imaged sample for further high-resolution focused ion beam/scanning electron microscope tomography. The subcellular structures of synapses obtained with the X-ray technique matched the electron microscopy data, demonstrating that PXCT is a powerful scanning method for tissue volumes of more than 60 cells and sensitive enough for identification of regions as small as 200 nm, which remain available for further structural and biochemical investigations.

Direct capsid labeling of infectious HIV-1 by genetic code expansion allows detection of largely complete nuclear capsids and suggests nuclear entry of HIV-1 complexes via common routes

The cone-shaped mature HIV-1 capsid is the main orchestrator of early viral replication. After cytosolic entry, it transports the viral replication complex along microtubules towards the nucleus. Capsid uncoating from the viral genome apparently occurs beyond the nuclear pore. Observation of post-entry events via microscopic detection of HIV-1 capsid protein (CA) is challenging, since epitope shielding limits immunodetection, and the genetic fragility of CA hampers other labeling approaches. Here, we present a minimally invasive strategy based on genetic code expansion and click chemistry that allows for site-directed fluorescent labeling of HIV-1 CA, while retaining virus morphology and infectivity. Thereby, we could directly visualize virions and subviral complexes using advanced microscopy, including nanoscopy and correlative imaging. Quantification of signal intensities of subviral complexes showed that the amount of CA associated with nuclear complexes in HeLa-derived cells and primary T cells is consistent with a complete capsid and revealed that treatment with the small molecule inhibitor PF74 did not result in capsid dissociation from nuclear complexes. Cone-shaped objects detected in the nucleus by electron tomography were clearly identified as capsid-derived structures by correlative microscopy. High-resolution imaging revealed dose-dependent clustering of nuclear capsids, suggesting that incoming particles may follow common entry routes.

Coming together—symbiont acquisition and early development in deep-sea bathymodioline mussels

How and when symbionts are acquired by their animal hosts has a profound impact on the ecology and evolution of the symbiosis. Understanding symbiont acquisition is particularly challenging in deep-sea organisms because early life stages are so rarely found. Here, we collected early developmental stages of three deep-sea bathymodioline species from different habitats to identify when these acquire their symbionts and how their body plan adapts to a symbiotic lifestyle. These mussels gain their nutrition from chemosynthetic bacteria, allowing them to thrive at deep-sea vents and seeps worldwide. Correlative imaging analyses using synchrotron-radiation based microtomography together with light, fluorescence and electron microscopy revealed that the pediveliger larvae were aposymbiotic. Symbiont colonization began during metamorphosis from a planktonic to a benthic lifestyle, with the symbionts rapidly colonizing first the gills, the symbiotic organ of adults, followed by all other epithelia of their hosts. Once symbiont densities in plantigrades reached those of adults, the host's intestine changed from the looped anatomy typical for bivalves to a straightened form. Within the Mytilidae, this morphological change appears to be specific to Bathymodiolus and Gigantidas , and is probably linked to the decrease in the importance of filter feeding when these mussels switch to gaining their nutrition largely from their symbionts.