Computed tomography of cryogenic biological specimens based on X-ray microscopic images

Weiss, Daniel H.; Schneider, Gerd; Niemann, B.; Guttmann, Peter; Rudolph, D.; Schmahl, G.

doi:10.1016/s0304-3991(00)00034-6

Cited by 252 publications

(137 citation statements)

References 9 publications

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“…As long as the thickness of the object does not exceed the depth of focus of the objective lens of the microscope, transmission images' approximate projections through the object, which can be used to obtain three-dimensional reconstructions by the method of tomography. The high contrast enables images to be obtained at a tolerable dose at a resolution approaching 30 nm, but further improvements in resolution for this wavelength range require increasing the numerical aperture of the objective, which would reduce the depth of focus to substantially less than 1 μm, thinner than most cells [4].…”

Section: Introductionmentioning

confidence: 99%

“…Current X-ray microscopy techniques operate near this limit. Images of 5-μm-thick cryogenically cooled cells were obtained by water-window absorption-based imaging at about 30 nm resolution [2,4] at about 1 GGy dose. Cryogenically cooled cells of 18 μm thickness were imaged by ptychography at 6.2 keV photon energy, achieving 180-nm resolution at 670 kGy [21], and a 5-μm-wide alga cell was imaged in a similar fashion at 5.2 keV photon energy to achieve 18-nm resolution at 29 MGy [22].…”

Section: Introductionmentioning

confidence: 99%

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Dose efficient Compton X-ray microscopy

Villanueva-Pérez¹,

Bajt²,

Chapman³

2018

Optica

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X-ray imaging techniques have proven invaluable to study biological systems at high resolution due to the penetration power and short wavelength of this radiation. In practice, the resolution and sensitivity of current X-ray imaging techniques are not limited by the performance of optics or image-recovery methods but by radiation damage. We propose the use of Compton (inelastic) X-ray scattering for high-resolution cellular imaging and provide a study of a scanning microscope geometry that requires a dose to achieve a given resolution that is three orders of magnitude lower than for coherent (elastic) scattering. We find that the dose per imaging signal is minimized at a photon energy of 64 keV. This corresponds to a short enough wavelength (0.02 nm) to provide nanometer transverse resolution and micrometer depth of field for tomographic imaging of whole cells. The microscope could be implemented at future high-energy and high-brightness synchrotron-radiation facilities to provide images of unsectioned and unlabeled cells in their native conditions at enough detail to bridge the techniques of super-resolution optical microscopy and cryo-electron microscopy. © 2018 Optical Society of America under the terms of the OSA Open Access Publishing Agreement OCIS codes: (110.7440) X-ray imaging; (340.7460) X-ray microscopy; (170.3880) Medical and biological imaging.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Dose efficient Compton X-ray microscopy

Villanueva-Pérez¹,

Bajt²,

Chapman³

2018

Optica

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“…Synchrotron radiation-based X-ray tomography can traverse the sample thickness limits of electron microscopy and resolution restrictions of CLSM [12,17,18]. Several groups demonstrated high resolution 3D imaging of biological objects of yeast [16,17,19], lymphocyte [18], virus membranes [20], bacteria [21,22], eukaryotic chromosomes [23] and algae [24] in the soft X-ray in the "water window" spectral region (wavelength region: 2.34 nm -4.44 nm, photon energy region: 0.28 -0.53 KeV) [16,17,20] and hard X-ray spectral region (wavelength region: 0.01 nm -0.62 nm, photon energy region: 2 -100 KeV) [21,23]. Compared with soft X-ray microscopy, hard X-ray microscopy has relatively large penetration depths, focal lengths and depths of focus, which has advantages for 3D reconstruction of much thicker cells and tissues [25,26].…”

Section: Introductionmentioning

confidence: 99%

The analysis of microscopy imaging on liquid crystalline components of the cell nucleus

Sun¹,

Liu²,

Dai³

et al. 2012

JBiSE

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Dinoflagellates nuclei allow for liquid crystalline characterization without core histones. In this study, nuclei were isolated from the athecate Karenia dinoflagellate species with minimum destruction to their native structure during preparation procedures. The liquid crystalline nuclei were studied by microscopy techniques of Metripol birefringence microscopy, Confocal Laser Scanning Microscopy (CLSM) and synchrotron radiation-based hard X-ray Microscopy with computed tomography, respectively. The 3D reconstruction techniques of hard X-ray tomography and CLSM were also discussed. The important biophysical parameters of the interspaces between chromosomes, nuclear surface areas and chromosome-occupied volumes were calculated from a 3D rendering of a reconstructed nucleus. The results of calculated average chromosomal DNA concentration of dinoflagellate was consistent with the concentration which can spontaneously assemble into the cholesteric liquid crystal phase in vitro.

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“…Transmission x-ray microscopy in the spectral range of the "water window" region (λ = 2.3 -4.4 nm) is a powerful tool for the investigation of biological and mineralogical samples, including, e.g., tomographic studies of cryogenic cells [1][2][3][4][5][6][7][8] and spectromicroscopic analysis of soils due to the element-specific contrast [9]. Spatial resolutions of 10 nm have been achieved [10] making use of Fresnel zone plates as highly magnifying objectives.…”

Section: Introductionmentioning

confidence: 99%