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Related Concept Videos

Introduction to Membrane Traffic01:44

Introduction to Membrane Traffic

The ER, Golgi apparatus, endosomes, and lysosomes work in tandem to modify, sort, and package proteins and lipids. An integrated membrane trafficking network facilitates the back and forth shuttling of molecules within different organelles in the same cell or across the cell membrane.
The transport of soluble and membrane proteins is mediated by transport vesicles that collect cargo from one cellular compartment and deliver it to another by fusing with the target organelle membrane. The Rab...
Introduction to Membrane Traffic01:44

Introduction to Membrane Traffic

The ER, Golgi apparatus, endosomes, and lysosomes work in tandem to modify, sort, and package proteins and lipids. An integrated membrane trafficking network facilitates the back and forth shuttling of molecules within different organelles in the same cell or across the cell membrane.
The transport of soluble and membrane proteins is mediated by transport vesicles that collect cargo from one cellular compartment and deliver it to another by fusing with the target organelle membrane. The Rab...

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High-resolution Spatiotemporal Analysis of Receptor Dynamics by Single-molecule Fluorescence Microscopy
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Published on: July 25, 2014

Intracellular membrane traffic at high resolution.

Jan R T van Weering1, Edward Brown, Thomas H Sharp

  • 1Department of Biochemistry, School of Medical Sciences, University of Bristol, University Walk, Bristol BS81TD, United Kingdom.

Methods in Cell Biology
|September 28, 2010
PubMed
Summary
This summary is machine-generated.

Electron microscopy (EM) visualizes cellular processes at nanometer resolution. This review details advanced EM techniques for studying intracellular membrane traffic, particularly within the endosomal system.

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Area of Science:

  • Cell Biology
  • Microscopy

Background:

  • Membrane traffic is vital for cell homeostasis.
  • Key trafficking steps occur in organelle microdomains requiring high-resolution analysis.
  • Electron microscopy (EM) provides detailed subcellular morphology.

Purpose of the Study:

  • To review the application of EM techniques for studying intracellular membrane traffic.
  • To focus on the endosomal system and its molecular mechanisms.
  • To provide practical protocols for advanced EM methods.

Main Methods:

  • Review of advanced EM techniques including cryo-immuno-EM, correlative light electron microscopy (CLEM), and 3D electron tomography.
  • Detailed protocol for immunolabeling on Lowicryl sections of high-pressure frozen cells.
  • Description of a simple CLEM method for membrane trafficking studies.

Main Results:

  • EM techniques have evolved from morphological analysis to advanced applications.
  • Specific protocols are provided for immunolabeling and CLEM.
  • These methods enable detailed molecular analysis of membrane traffic.

Conclusions:

  • Advanced EM techniques are crucial for understanding intracellular membrane traffic.
  • These methods offer high-resolution insights into endosomal sorting and general membrane dynamics.
  • EM-based approaches significantly expand our knowledge of molecular mechanisms in cell biology.