Three-dimensional true FISP for high-resolution imaging of the whole brain

Abstract Background Organoids are a reliable model used in the study of human brain development and under pathological conditions. However, current methods for brain organoid culture generate tissues that range from 0.5 to 2 mm of size, which need to be constantly agitated to allow proper oxygenation. The culture conditions are, therefore, not suitable for whole-brain organoid live imaging, required to study developmental processes and disease progression within physiologically relevant time frames (i.e. days, weeks, months). Results Here we designed 3D-printed microplate inserts adaptable to standard 24 multi-well plates, which allow the growth of multiple organoids in pre-defined and fixed XYZ coordinates. This innovation facilitates high-resolution imaging of whole-cerebral organoids, allowing precise assessment of organoid growth and morphology, as well as cell tracking within the organoids, over long periods. We applied this technology to track neocortex development through neuronal progenitors in brain organoids, as well as the movement of patient-derived glioblastoma stem cells within healthy brain organoids. Conclusions This new bioengineering platform constitutes a significant advance that permits long term detailed analysis of whole-brain organoids using multimodal inverted fluorescence microscopy.

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Two- and Three-Dimensional High Resolution Imaging of Genetic Expression of Luciferase

Biomedical Topical Meeting ◽

10.1364/bio.2004.thf21 ◽

2004 ◽

Author(s):

B. Chance ◽

W. El-Deiry ◽

Y. Li ◽

L Zhou

Keyword(s):

High Resolution ◽

Three Dimensional ◽

High Resolution Imaging ◽

Genetic Expression ◽

Resolution Imaging

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

Scientific Reports ◽

10.1038/s41598-020-78150-3 ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Carla C. Polo ◽

Miriam H. Fonseca-Alaniz ◽

Jian-Hua Chen ◽

Axel Ekman ◽

Gerry McDermott ◽

...

Keyword(s):

High Resolution ◽

Three Dimensional ◽

Linear Attenuation Coefficient ◽

High Resolution Imaging ◽

Mitochondrial Matrix ◽

Morphological Parameter ◽

Mitochondrial Membranes ◽

X Ray ◽

Resolution Imaging ◽

Neuronal Disorders

AbstractMitochondria 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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