Native Cellular Ultrastructure
Native Cellular Ultrastructure - Related Videos
12:03
Blue Native Polyacrylamide Gel Electrophoresis (BN-PAGE) for Analysis of Multiprotein Complexes from Cellular Lysates
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Cited by 92 •
2011
In this video, we describe the characterization of multiprotein complexes (MPCs) by blue native polyacrylamide gel electrophoresis (BN-PAGE). In a first dimension, dialyzed cellular lysates are separated by BN-PAGE to identify individual MPCs. In a second dimension SDS-PAGE, MPCs of interest are further subdivided to analyze their constituents by immunoblotting.
10:49
Identification of Small Molecule-binding Proteins in a Native Cellular Environment by Live-cell Photoaffinity Labeling
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Cited by 4 •
2016
We describe here a method for identification of small molecule-binding proteins using photoaffinity labeling. The advantage of this technique is that binding and covalent labeling of the target proteins occurs within the live cellular environment, removing the risk of disrupting native protein structure and binding conditions upon cell lysis.
02:53
Ultrastructural Analysis of a Mouse Brain Section Using Transmission Electron Microscopy
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2025
Source: Lutz, D., et.al. Assessment of Ultrastructural Neuroplasticity Parameters After In Utero Transduction of the Developing Mouse Brain and Spinal Cord. J. Vis. Exp. (2019).This video demonstrates the preparation and transmission electron microscopy (TEM) analysis of a brain section from a mouse pup. The protocol starts with osmium tetroxide-fixed tissue embedded in resin, followed by precise trimming to isolate the region of interest and ultramicrotomy to produce semithin and ultrathin...
09:02
Split-BioID — Proteomic Analysis of Context-specific Protein Complexes in Their Native Cellular Environment
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Cited by 12 •
2018
We provide a step-by-step protocol for split-BioID, a protein fragments-complementation assay based on the proximity-labeling technique BioID. Activated on the interaction of two given proteins, it allows the proteomics analysis of context-dependent protein complexes in their native cellular environment. The method is simple, cost-effective and only requires standard laboratory equipment.
05:48
Rapid Generation of Amyloid from Native Proteins In vitro
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Cited by 10 •
2013
Proteins can either adopt a native structure or misfold into insoluble amyloid. Conditions that favor the misfolding pathway lead to the formation of different types of amyloid fibrils. The methods described here allow rapid conversion of native proteins into amyloid in vitro.
FAQs
Rapid freezing converts cellular water into a vitrified state, stabilizing it without the structural disruption associated with chemical fixation or dehydration. This preservation keeps membranes, organelles, cytoskeletal elements, and protein assemblies positioned close to their native cellular relationships. As a result, images can be interpreted as evidence of organization within near-natural neuronal contexts rather than as isolated processed components.
Membranes, organelles, cytoskeletal elements, and protein assemblies provide complementary structural information. Membranes define cellular compartments, cytoskeletal elements reveal internal organization, and protein assemblies contribute fine-scale molecular detail. Considering these features together helps connect the placement of cellular components with how synapses, axons, dendrites, and intracellular compartments are arranged and interact.
Three-dimensional tomograms place observed structures within a volumetric context, rather than limiting interpretation to a single electron image. This makes it possible to examine how cellular components are arranged relative to one another and to follow relationships across complex neuronal regions. Such spatial information strengthens comparisons between molecular architecture and the organization of synapses, axons, dendrites, and intracellular compartments.
By avoiding chemical fixation and dehydration, the approach preserves cellular material under near-natural conditions before imaging. That distinction matters because the resulting structural record is intended to retain the organization of membranes, organelles, cytoskeletal elements, and protein assemblies. It therefore supports a different interpretation from methods that first chemically stabilize or dehydrate the specimen.
The workflow begins with rapid freezing to vitrify cellular water and stabilize the specimen. Cryogenic electron microscopy then produces images of the preserved material, and those images are reconstructed into three-dimensional tomograms. This sequence links physical preservation to volumetric structural analysis, allowing investigators to examine native cellular organization within neuronal samples.
In neuroscience, the method can be directed toward synapses, axons, dendrites, and intracellular compartments. Examining these structures in their preserved cellular arrangement helps researchers study how neuronal regions are positioned and how they interact. The resulting architecture provides a structural basis for interpreting neural organization, rather than treating each compartment as an unrelated isolated feature.
Native Cellular Ultrastructure can connect molecular architecture with cellular activity by showing where fine structural elements occur within neuronal cells. This relationship may help clarify mechanisms of neural communication and development, while also informing investigations of disease. Its value lies in relating detailed physical organization to broader questions about how brain cells function and change.