Quantitative imaging of <i>Candida utilis</i> and its organelles by soft X‐ray Nano‐CT

Liu, J.; Li, F.; Chen, L.; Guan, Yong; Tian, Li-Jiao; Xiong, Ying; Liu, G.; Tian, Yangchao

doi:10.1111/jmi.12650

Cited by 25 publications

(13 citation statements)

References 34 publications

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“…From their LAC histogram ( fig. S11), the average LAC value of the Iod-CBT-NPs in the above 3D rendering image was calculated to be 0.182 ± 0.078 m −1 by Gaussian fitting (32,33). In contrast, the average LAC value of the background of above 3D rendering image (i.e., buffer salts) was much lower (0.016 ± 2.043 × 10 −4 m −1 ) ( fig.…”

Section: Ntr-triggered Iod-cbt-np Formation In Vitromentioning

confidence: 96%

Directly observing intracellular nanoparticle formation with nanocomputed tomography

et al. 2020

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Directly observing intracellular nanostructure formation remains challenging. In this work, using a rationally designed small-molecule 4-nitrobenzyl carbamate–Cys(SEt)-Asp-Asp-Phe(iodine)–2-cyano-benzothiazole (NBC-Iod-CBT), we directly observed intracellular nanoparticle formation with nanocomputed tomography (nano-CT). In vitro, upon glutathione reduction and nitroreductase (NTR) cleavage, NBC-Iod-CBT undergoes a CBT-Cys click condensation reaction to self-assemble nanoparticles Iod-CBT-NPs with an average linear absorption coefficient (LAC) value of 0.182 ± 0.078 μm−1 to x-ray. Nano-CT imaging of the NBC-Iod-CBT–treated, NTR-overexpressing HeLa cells showed the existence of Iod-CBT-NPs in their cytoplasm with an average LAC value of 0.172 ± 0.032 μm−1. We anticipate that our strategy could help people to deeply understand the formation mechanism of intracellular nanostructures in the near future.

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Section: Ntr-triggered Iod-cbt-np Formation In Vitromentioning

confidence: 96%

Directly observing intracellular nanoparticle formation with nanocomputed tomography

et al. 2020

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“…On the other hand, the use of X-ray based imaging can provide information about thick and large area biological samples and under cryogenic conditions to avoid the radiation damage, for instance the modalities of X-ptychography of vegetal tissues 18 and soft X-rays of whole cells 19 . Mitochondria has been already explored by X-ray imaging to be identified in the cellular context of wild-type single cells 19,20 , in cancer cells 21,22 and isolated from the cells aiming to unveil its internal structure 23 . However, there are no reports of X-ray imaging-based analysis focusing on extracting 3D morphological and quantitative information about the OMM and IMM organization.…”

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confidence: 99%

Three-dimensional imaging of mitochondrial cristae complexity using cryo-soft X-ray tomography

Polo

Fonseca‐Alaniz

Chen

et al. 2020

Sci Rep

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Mitochondria are dynamic organelles that change morphology to adapt to cellular energetic demands under both physiological and stress conditions. Cardiomyopathies and neuronal disorders are associated with structure-related dysfunction in mitochondria, but three-dimensional characterizations of the organelles are still lacking. In this study, we combined high-resolution imaging and 3D electron density information provided by cryo-soft X-ray tomography to characterize mitochondria cristae morphology isolated from murine. Using the linear attenuation coefficient, the mitochondria were identified (0.247 ± 0.04 µm−1) presenting average dimensions of 0.90 ± 0.20 µm in length and 0.63 ± 0.12 µm in width. The internal mitochondria structure was successfully identified by reaching up the limit of spatial resolution of 35 nm. The internal mitochondrial membranes invagination (cristae) complexity was calculated by the mitochondrial complexity index (MCI) providing quantitative and morphological information of mitochondria larger than 0.90 mm in length. The segmentation to visualize the cristae invaginations into the mitochondrial matrix was possible in mitochondria with MCI ≥ 7. Altogether, we demonstrated that the MCI is a valuable quantitative morphological parameter to evaluate cristae modelling and can be applied to compare healthy and disease state associated to mitochondria morphology.

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“…The LAC values represent the soft X-ray absorption intensity, and can be calculated using the following equation (1) (39): where I 0 is the initial X-ray intensity and I is the output X-ray intensity. μ (x) is the local LAC value, which can be calculated by an iterative reconstruction using the TV-SART algorithm (40). The LAC value was determined based on the mass absorption coefficient and the density of the composition, as defined in the following equation (2): where μ is the LAC, μ m is the mass absorption coefficient, and ρ is the density of the composition.…”

Section: Discussionmentioning

confidence: 99%

“…The reconstruction results of the TV-SART algorithm were evaluated using the LAC values. The LAC represents the inherent ability of a substance for absorption the soft x-ray (39), and the images depicting the LAC were segmented to identify the different organelle structures and chemical composition (40, 41). Amira software was used to segment the reconstructed anammox bacteria cell, calculate the LAC values, measure the number of pixels of the anammoxosome organelle and anammox bacteria cell, and to create movies (26, 42).…”

Section: Methodsmentioning

confidence: 99%

Quantitative three-dimensional nondestructive imaging of whole anaerobic ammonium-oxidizing bacteria

Peng

Guan

Liu

et al. 2019

Preprint

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AbstractAnaerobic ammonium-oxidizing (anammox) bacteria play a key role in the global nitrogen cycle and the treatment of nitrogenous wastewater. These functions are closely related to the unique biophysical structure of anammox bacteria. However, the research on the biophysical ultrastructure of intact anammox bacteria is lacking. In this study, in-situ three-dimensional nondestructive ultrastructure imaging of whole anammox cell was performed using synchrotron soft X-ray nano-computed tomography and the total variation-based simultaneous algebraic reconstruction technique (TV-SART). Statistical and quantitative analyses of the ultrastructures of intact anammox bacteria were performed. The linear absorption coefficient values of the ultrastructures of anammox bacteria were calculated and the asymmetric structure of the anammox bacteria was quantified. On this basis, the shape adaptation of the anammox bacteria responses to Fe2+ were explored, and the underlying regulation mechanism of Fe2+ on anammox bacteria was explored. Furthermore, a promising method to study the biophysical properties of cells in different environments and engineering processes was proposed.Graphical AbstractStatement of SignificanceAnaerobic ammonium-oxidizing (anammox) bacteria play key role in global nitrogen cycle, and this physiological function depends on the unique morphology of anammox bacteria. In this study, synchrotron soft-X ray imaging technique coupled with simultaneous algebraic reconstruction technique with total variation (SART-TV) algorithm were performed to quantify the three-dimensional ultrastructure of the whole anammox bacteria for the first time. On this basis, the shape adaptation and mechanism of the anammox bacteria responses to Fe2+ were explored and a promising method for detecting the physiological properties of anammox bacteria was proposed.

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Quantitative imaging of Candida utilis and its organelles by soft X‐ray Nano‐CT

Cited by 25 publications

References 34 publications

Directly observing intracellular nanoparticle formation with nanocomputed tomography

Directly observing intracellular nanoparticle formation with nanocomputed tomography

Three-dimensional imaging of mitochondrial cristae complexity using cryo-soft X-ray tomography

Quantitative three-dimensional nondestructive imaging of whole anaerobic ammonium-oxidizing bacteria

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