a-3d-cartographic-description-cell-cryo-soft-x-ray

A 3D Cartographic Description of the Cell by Cryo Soft X-ray Tomography

Imaging techniques are fundamental in order to understand cell organization and machinery in biological research and the related fields. Among these ...

cryo-structured-illumination-microscopic-data-collection-from

Cryo-Structured Illumination Microscopic Data Collection from Cryogenically Preserved Cells

Three-dimensional (3D) structured illumination microscopy (SIM) allows imaging of fluorescently labelled cellular structures at higher resolution than ...

inext-discovery-instruct-eric-integrating-high-end-services-for

iNEXT-Discovery and Instruct-ERIC: Integrating High-End Services for Translational Research in Structural Biology

ARTICLES DISCUSSED:
Ahmad, M. U. D. et al. Nano-differential scanning fluorimetry for screening in fragment-based lead discovery. Journal of Visualized ...

Alba Light Source, Cerdanyola del Valles, Spain

Beamline 09 - MISTRAL

ALBA Light Source

The MISTRAL beamline is one of the seven phase-I beamlines at the ALBA synchrotron light source (Barcelona, Spain) that will be opened to users at the end of 2010. MISTRAL will be devoted to cryotomography in the water window and multi-keV spectral regions for biological applications. The optics design consists of a plane-grating monochromator that has been implemented using variable-line-spacing gratings to fulfil the requirements of X-ray microscopy using a reflective condenser. For instance, a fixed-focus condition independent of the included angle, constant magnification as well as coma and spherical aberration corrections are achieved with this system. The reported design is of wider use.

A soft X-ray beamline for transmission X-ray microscopy at ALBA.

Recent advances in nanoparticle design have generated new possibilities for nano-biotechnology and nano-medicine. Here we used cryo-soft X-ray tomography (cryo-SXT) to collect comprehensive three-dimensional (3D) data to characterise the interaction of superparamagnetic iron oxide nanoparticles (SPION) with a breast cancer cell line.

Cryo-soft X-ray tomography as a quantitative three-dimensional tool to model nanoparticle:cell interaction.

Correlative cryo-soft X-ray tomography of cells.

3D correlative microscopy methods have revolutionized biomedical research, allowing the acquisition of multidimensional information to gain an in-depth understanding of biological systems. With the advent of relevant cryo-preservation methods, correlative imaging of cryogenically preserved samples has led to nanometer resolution imaging (2-50 nm) under harsh imaging regimes such as electron and soft X-ray tomography. These methods have now been combined with conventional and super-resolution fluorescence imaging at cryogenic temperatures to augment information content from a given sample, resulting in the immediate requirement for protocols that facilitate hassle-free, unambiguous cross-correlation between microscopes. We present here sample preparation strategies and a direct comparison of different working fiducialization regimes that facilitate 3D correlation of cryo-structured illumination microscopy and cryo-soft X-ray tomography. Our protocol has been tested at two synchrotron beamlines (B24 at Diamond Light Source in the UK and BL09 Mistral at ALBA in Spain) and has led to the development of a decision aid that facilitates experimental design with the strategic use of markers based on project requirements. This protocol takes between 1.5 h and 3.5 d to complete, depending on the cell populations used (adherent cells may require several days to grow on sample carriers).

Eva Pereiro

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Alba Light Source, Cerdanyola del Valles, Spain

A 3D Cartographic Description of the Cell by Cryo Soft X-ray Tomography

Cryo-Structured Illumination Microscopic Data Collection from Cryogenically Preserved Cells

iNEXT-Discovery and Instruct-ERIC: Integrating High-End Services for Translational Research in Structural Biology

MISTRAL beamline