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We applied ASEM to three-dimensional FIB-SEM images of cells prepared by either CF or HPFS. We note that while cellular samples have traditionally been processed by chemical fixation (CF) and staining at room temperature, HPFS at cryogenic temperatures (as was the case for the COSEM Project data) yields a substantial increase in the preservation of many complex cellular features. We illustrate here the utility of ASEM by describing the results of its application to data from several types of cells, including FIB-SEM datasets made publicly available by the COSEM Project ( Heinrich et al., 2021). The most common strategy used by the COSEM project involved training with ground truth annotations from multiple classes of objects at the same time, typically at a high computational cost (500,000 or more training iterations Heinrich et al., 2021). A pipeline created by the COSEM project ( Heinrich et al., 2021) enables automated whole-cell simultaneous segmentation of up to 35 organelles from relatively sparse but very precise 3D ground truth annotations from FIB-SEM data of cells prepared by high-pressure freezing and freeze substitution (HPFS), obtained at 3–5 nm voxel size with approximately isotropic resolution. However, it was conceived for anisotropic data and therefore has conceptual limitations when applied to 3D isotropic FIBSEM data. CDeep3M was the first project using cloud computing and achieves good results on mostly large-size organelles or clusters of smaller-size vesicles. Examples using deep convolutional networks for data with anisotropic resolution include DeepEM3D ( Zeng et al., 2017) and CDeep3M ( Haberl et al., 2018), for segmentation of mitochondria and Golgi apparatus with extensive post-processing ( Žerovnik Mekuč et al., 2020 Žerovnik Mekuč et al., 2022), as well as cell organelle segmentation in quasi-isotropic FIB-SEM data of beta cells ( Müller et al., 2021). The widespread success of machine learning in bioimage analysis has recently inspired the application of deep learning approaches to automated segmentation. We uncovered a wide range of membrane–nuclear pore diameters within a single cell and derived morphological metrics from clathrin-coated pits and vesicles, consistent with the classical constant-growth assembly model. We identified mitochondria, Golgi apparatus, endoplasmic reticulum, nuclear pore complexes, caveolae, clathrin-coated pits, and vesicles imaged by focused ion beam scanning electron microscopy. Model generalization was improved with a rapid, computationally effective strategy to refine a trained model by including a few additional annotations. We obtained dedicated models for each structure based on a small number of sparsely annotated ground truth images from only one or two cells. We resolved this bottleneck with automated segmentation of intracellular substructures in electron microscopy (ASEM), a new pipeline to train a convolutional neural network to detect structures of a wide range in size and complexity. Identification of intracellular structures is a laborious process limiting the effective use of this potentially powerful tool. In an apparent suicide, the photographer was found dead with a plastic bag over his head on Novemin Paris, France.Volume electron microscopy is an important imaging modality in contemporary cell biology. In 2016, the photographer was accused of rape by four separate women who had served as former models. Similar to the work of Jock Sturges and Sally Mann, Hamilton’s erotic subject matter caused complaints of indecency over the decades. He published numerous photobooks during his life, The Age of Innocence (1995) being one of his most popular.

Nn model ru sets movie#

In 1979, his film Laura was released in France-the movie blends melodrama with soft-core pornography, as wells as Hamilton’s hallmark soft focus. Born on Apin London, United Kingdom, the artist initially found work as an art director, then a commercial photographer in the 1950s and 60s in France. Using a photographic technique akin to painting, he has acknowledged the influence of Lucas Cranach on his style. Often controversial, the images evoke an effect of dreaminess through soft focus and varied grain. David Hamilton was a British photographer and film director known for his nude photographs of adolescent girls.