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Cryo-soft X-ray tomography: a journey into the world of the native-state cell.

Carzaniga R, Domart MC, Collinson LM, Duke E - Protoplasma (2013)

Bottom Line: One of the ultimate aims of imaging in biology is to achieve molecular localisation in the context of the structure of cells in their native state.Here, we review the current state of the art in cryo-soft X-ray tomography (cryo-SXT), which is the only imaging modality that can provide nanoscale 3D information from cryo-preserved, unstained, whole cells thicker than 1 μm.Correlative cryo-fluorescence and cryo-SXT adds functional information to structure, enabling studies of cellular events that cannot be captured using light, electron or X-ray microscopes alone.

View Article: PubMed Central - PubMed

Affiliation: Electron Microscopy Unit, London Research Institute, Cancer Research UK, London, WC2A 3LY, UK.

ABSTRACT
One of the ultimate aims of imaging in biology is to achieve molecular localisation in the context of the structure of cells in their native state. Here, we review the current state of the art in cryo-soft X-ray tomography (cryo-SXT), which is the only imaging modality that can provide nanoscale 3D information from cryo-preserved, unstained, whole cells thicker than 1 μm. Correlative cryo-fluorescence and cryo-SXT adds functional information to structure, enabling studies of cellular events that cannot be captured using light, electron or X-ray microscopes alone.

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Related in: MedlinePlus

Zone plates for X-ray microscopy. Zone plates are made from concentric rings of material that focus the X-ray beam. The width of the outermost zone (Δrn) governs the maximum resolution achievable from data collected using the zone plate
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Fig3: Zone plates for X-ray microscopy. Zone plates are made from concentric rings of material that focus the X-ray beam. The width of the outermost zone (Δrn) governs the maximum resolution achievable from data collected using the zone plate

Mentions: In the transmission X-ray microscope, the X-rays are condensed onto the sample and collected using a diffractive, focusing optical element called a zone plate. Zone plates were invented by Rayleigh (unpublished) and the theory of their operation described by Soret (1875). Decades of research and development in both materials and manufacturing techniques were required to produce zone plates suitable for use with soft X-rays (Baez 1961). Formed from alternating opaque and transparent concentric rings (zones), it is the radius of the outermost ring that determines the resolution of the microscope (Fig. 3). Synchrotron-hosted soft X-ray microscopes routinely operate using two alternative zones plates, one at 40-nm resolution and the other at 25-nm resolution. Unfortunately, zone plates are incredibly inefficient optical elements, with the majority operating at around 10 % efficiency, particularly in the soft X-ray region. Given that the zone plate is fundamental to the operation of the X-ray microscope, it is not surprising that there are worldwide efforts to develop new techniques for zone plate fabrication.Fig. 3


Cryo-soft X-ray tomography: a journey into the world of the native-state cell.

Carzaniga R, Domart MC, Collinson LM, Duke E - Protoplasma (2013)

Zone plates for X-ray microscopy. Zone plates are made from concentric rings of material that focus the X-ray beam. The width of the outermost zone (Δrn) governs the maximum resolution achievable from data collected using the zone plate
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3927064&req=5

Fig3: Zone plates for X-ray microscopy. Zone plates are made from concentric rings of material that focus the X-ray beam. The width of the outermost zone (Δrn) governs the maximum resolution achievable from data collected using the zone plate
Mentions: In the transmission X-ray microscope, the X-rays are condensed onto the sample and collected using a diffractive, focusing optical element called a zone plate. Zone plates were invented by Rayleigh (unpublished) and the theory of their operation described by Soret (1875). Decades of research and development in both materials and manufacturing techniques were required to produce zone plates suitable for use with soft X-rays (Baez 1961). Formed from alternating opaque and transparent concentric rings (zones), it is the radius of the outermost ring that determines the resolution of the microscope (Fig. 3). Synchrotron-hosted soft X-ray microscopes routinely operate using two alternative zones plates, one at 40-nm resolution and the other at 25-nm resolution. Unfortunately, zone plates are incredibly inefficient optical elements, with the majority operating at around 10 % efficiency, particularly in the soft X-ray region. Given that the zone plate is fundamental to the operation of the X-ray microscope, it is not surprising that there are worldwide efforts to develop new techniques for zone plate fabrication.Fig. 3

Bottom Line: One of the ultimate aims of imaging in biology is to achieve molecular localisation in the context of the structure of cells in their native state.Here, we review the current state of the art in cryo-soft X-ray tomography (cryo-SXT), which is the only imaging modality that can provide nanoscale 3D information from cryo-preserved, unstained, whole cells thicker than 1 μm.Correlative cryo-fluorescence and cryo-SXT adds functional information to structure, enabling studies of cellular events that cannot be captured using light, electron or X-ray microscopes alone.

View Article: PubMed Central - PubMed

Affiliation: Electron Microscopy Unit, London Research Institute, Cancer Research UK, London, WC2A 3LY, UK.

ABSTRACT
One of the ultimate aims of imaging in biology is to achieve molecular localisation in the context of the structure of cells in their native state. Here, we review the current state of the art in cryo-soft X-ray tomography (cryo-SXT), which is the only imaging modality that can provide nanoscale 3D information from cryo-preserved, unstained, whole cells thicker than 1 μm. Correlative cryo-fluorescence and cryo-SXT adds functional information to structure, enabling studies of cellular events that cannot be captured using light, electron or X-ray microscopes alone.

Show MeSH
Related in: MedlinePlus