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1.
PLoS One ; 13(10): e0206172, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30352088

RESUMO

Serial section transmission electron microscopy (ssTEM) is the most promising tool for investigating the three-dimensional anatomy of the brain with nanometer resolution. Yet as the field progresses to larger volumes of brain tissue, new methods for high-yield, low-cost, and high-throughput serial sectioning are required. Here, we introduce LASSO (Loop-based Automated Serial Sectioning Operation), in which serial sections are processed in "batches." Batches are quantized groups of individual sections that, in LASSO, are cut with a diamond knife, picked up from an attached waterboat, and placed onto microfabricated TEM substrates using rapid, accurate, and repeatable robotic tools. Additionally, we introduce mathematical models for ssTEM with respect to yield, throughput, and cost to access ssTEM scalability. To validate the method experimentally, we processed 729 serial sections of human brain tissue (~40 nm x 1 mm x 1 mm). Section yield was 727/729 (99.7%). Sections were placed accurately and repeatably (x-direction: -20 ± 110 µm (1 s.d.), y-direction: 60 ± 150 µm (1 s.d.)) with a mean cycle time of 43 s ± 12 s (1 s.d.). High-magnification (2.5 nm/px) TEM imaging was conducted to measure the image quality. We report no significant distortion, information loss, or substrate-derived artifacts in the TEM images. Quantitatively, the edge spread function across vesicle edges and image contrast were comparable, suggesting that LASSO does not negatively affect image quality. In total, LASSO compares favorably with traditional serial sectioning methods with respect to throughput, yield, and cost for large-scale experiments, and represents a flexible, scalable, and accessible technology platform to enable the next generation of neuroanatomical studies.


Assuntos
Processamento de Imagem Assistida por Computador/métodos , Imageamento Tridimensional/métodos , Microscopia Eletrônica de Transmissão/métodos , Neuroanatomia/métodos , Encéfalo/anatomia & histologia , Encéfalo/ultraestrutura , Humanos , Reprodutibilidade dos Testes
2.
Science ; 355(6327): 804, 2017 Feb 24.
Artigo em Inglês | MEDLINE | ID: mdl-28232543
3.
Proc Natl Acad Sci U S A ; 108(19): 7669-73, 2011 May 10.
Artigo em Inglês | MEDLINE | ID: mdl-21518911

RESUMO

Why does a single fire ant Solenopsis invicta struggle in water, whereas a group can float effortlessly for days? We use time-lapse photography to investigate how fire ants S. invicta link their bodies together to build waterproof rafts. Although water repellency in nature has been previously viewed as a static material property of plant leaves and insect cuticles, we here demonstrate a self-assembled hydrophobic surface. We find that ants can considerably enhance their water repellency by linking their bodies together, a process analogous to the weaving of a waterproof fabric. We present a model for the rate of raft construction based on observations of ant trajectories atop the raft. Central to the construction process is the trapping of ants at the raft edge by their neighbors, suggesting that some "cooperative" behaviors may rely upon coercion.


Assuntos
Formigas/fisiologia , Comportamento Animal , Animais , Formigas/anatomia & histologia , Fenômenos Biofísicos , Inundações , Interações Hidrofóbicas e Hidrofílicas , Microscopia Eletrônica de Varredura , Modelos Biológicos , Comportamento Social , Propriedades de Superfície , Imagem com Lapso de Tempo , Gravação em Vídeo , Água
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