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Non-separate Mouse Sclerochoroid/RPE/Retina Staining and Whole Mount for the Integral Observation of Subretinal Layer

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Cryo-transmission Electron Microscopy of Outer-inner Membrane Vesicles Naturally Secreted by Gram-negative Pathogenic Bacteria

Authors: Lidia Delgado, Nicolás Baeza, Carla Pérez-Cruz, Carmen López-Iglesias and Elena Mercadé, date: 09/20/2019, view: 2228, Q&A: 0

[Abstract] A protocol was developed to visualize and analyze the structure of membrane vesicles (MVs) from Gram-negative bacteria. It is now accepted that these micrometric spherical vesicles are commonly produced by cells from all three domains of life, so the protocol could be useful in the study of vesicles produced by eukaryotes and archaea as well as ...

Electron Microscopy Sample Preparation Protocol Enabling Nano-to-mesoscopic Mapping of Cellular Connectomes and Their Habitats in Human Tissues and Organs

Authors: Lucy Ngo, Anton D. Nathanson, Tomasz Garbowski, Ulf Knothe, Dirk Zeidler and Melissa L. Knothe Tate, date: 07/20/2019, view: 3708, Q&A: 0

[Abstract] Multibeam scanning electron microscopy (multiSEM) provides a technical platform for seamless nano-to-mesoscale mapping of cells in human tissues and organs, which is a major new initiative of the U.S. National Institutes of Health. Such cross-length-scale imaging is expected to provide unprecedented understanding of relationships between cellular ...

Preparation and Manipulation of Olfactory Epithelium Explant Cultures for Measurement of the Mechanical Tension of Individual Axons Using the Biomembrane Force Probe

Authors: Coralie Fouquet and Alain Trembleau, date: 04/20/2019, view: 2408, Q&A: 0

[Abstract] In this paper, we describe a protocol allowing measurement of the mechanical tension of individual axons grown ex vivo from neural tissue explants. This protocol was developed with primary cultures of olfactory epithelium explants from embryonic (E13.5) mice. It includes a detailed description of explant dissection and culture, as well as ...

Visualization of Plant Cell Wall Epitopes Using Immunogold Labeling for Electron Microscopy

Author: Mateusz Majda, date: 04/05/2019, view: 3567, Q&A: 0

[Abstract] Plant cell walls consist of different polysaccharides and structural proteins, which form a rigid layer located outside of the plasma membrane. The wall is also a very dynamic cell composite, which is characterized by complex polysaccharide interactions and various modifications during cell development. The visualization of cell wall components in ...

Electron Microscopic Detection of Proteins and Protein Complexes in Mammalian Cells Using APEX-tagged, Conditionally Stable Nanobodies

Authors: Thomas E. Hall, James Rae, Nicholas Ariotti and Robert G. Parton, date: 11/20/2018, view: 2845, Q&A: 0

[Abstract] The recent development of 3D electron microscopic techniques for cells and tissues has necessitated the development of new methods for the detection of proteins and protein-complexes in situ. The development of new genetic tags, such as the ascorbate peroxidase, APEX, for electron microscopic detection of tagged proteins has expanded the ...

Examining Autophagy in Plant by Transmission Electron Microscopy (TEM)

Authors: Xiyin Zheng, Chenguang Zhao and Yule Liu, date: 10/20/2018, view: 3278, Q&A: 0

[Abstract] In plants, macroautophagy, here referred as autophagy, is a degradation pathway during which the double-membrane structure named autophagosome engulfs the cargo and then fuses with vacuole for material recycling.

To investigate the process of autophagy, transmission electron microscopy (TEM) was used to monitor the ultrastructure of ...

Metal-tagging Transmission Electron Microscopy for Localisation of Tombusvirus Replication Compartments in Yeast

Authors: Isabel Fernández de Castro and Cristina Risco, date: 04/20/2018, view: 4114, Q&A: 0

[Abstract] Positive-stranded (+) RNA viruses are intracellular pathogens in humans, animals and plants. To build viral replicase complexes (VRCs) viruses manipulate lipid flows and reorganize subcellular membranes. Redesigned membranes concentrate viral and host factors and create an environment that facilitates the formation of VRCs within replication ...

Electron Tomography to Study the Three-dimensional Structure of Plasmodesmata in Plant Tissues–from High Pressure Freezing Preparation to Ultrathin Section Collection

Authors: William J. Nicolas, Emmanuelle Bayer and Lysiane Brocard, date: 01/05/2018, view: 5986, Q&A: 0

[Abstract] Plasmodesmata (PD) are nanometric (~20 nm wide) membrane lined pores encased in the cell walls of the adjacent plant cells. They allow the cells to exchange all types of molecules ranging from nutrients like sugar, hormones, to RNAs and various proteins. Unfortunately, they are also hijacked by phyto-viruses, enabling them to spread from ...

Immunogold Electron Microscopy of the Autophagosome Marker LC3

Authors: Domenico Mattoscio, Andrea Raimondi, Carlo Tacchetti and Susanna Chiocca, date: 12/20/2017, view: 8540, Q&A: 0

[Abstract] Even though autophagy was firstly observed by transmission electron microscopy already in the 1950s (reviewed in Eskelinen et al., 2011), nowadays this technique remains one of the most powerful systems to monitor autophagic processes. The autophagosome, an LC3-positive double membrane structures enclosing cellular materials, represents ...

Using Light and Electron Microscopy to Estimate Structural Variation in Thylakoid Membranes

Authors: Hatsumi Nozue and Kiyokazu Kametani, date: 12/05/2017, view: 6137, Q&A: 0

[Abstract] The shapes of chloroplasts and the architectures of internal thylakoid membranes are altered by growth and environmental changes (Lichtenthaler et al., 1981; Kutik, 1985; Terashima and Hikosaka, 1995). These morphological alterations proceed via transitional intermediates, during which dynamic and heterogeneous thylakoid membranes are ...

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