Hostname: page-component-586b7cd67f-l7hp2 Total loading time: 0 Render date: 2024-11-28T17:46:28.430Z Has data issue: false hasContentIssue false
  • English
  • Français

Dual Wavelength Evanescent Field Microscopy of Exocytosis and Endocytosis in Single Cells

Published online by Cambridge University Press:  02 July 2020

C. Merrifield ,
M. Feldman  and
W. Almers
C. Merrifield
Affiliation:
Vollum Institute, Oregon Health Sciences University, 3181 Sam Jackson Park Road, Portland, Oregon, 97201
M. Feldman
Affiliation:
Vollum Institute, Oregon Health Sciences University, 3181 Sam Jackson Park Road, Portland, Oregon, 97201
W. Almers
Affiliation:
Vollum Institute, Oregon Health Sciences University, 3181 Sam Jackson Park Road, Portland, Oregon, 97201
Get access

Abstract

The plasma membrane defines the outer limit of eukaryotic cells and it is here that cells interact with their environment through receptor proteins. Clearly the management of cell surface receptor concentration is critical for normal cellular function, and this is achieved through two key processes, exocytosis and endocytosis. During exocytosis transport vesicles containing both membrane receptors and soluble cargo fuse with the plasma membrane releasing cargo and delivering receptors to the cell surface. During clathrin mediated endocytosis receptors destined for internalisation become concentrated in clathrin coated pits before coat invagination and membrane fission. Since transferrin receptors are recycled and undergo numerous rounds of exocytosis and endocytosis by labeling transferrin receptor with an appropriate fluorophore it is possible to label both exocytic and endocytic organelles in the same cell. We have exploited this phenomenon to visualise the spatial and temporal organisation of constitutive exocytosis and endocytosis at the surface of single cells using dual wavelength evanescent field microscopy.

Type
Correlative Fluorescent Microscopy and Flow Cytometry Techniques (Organized by R. Smith)
Information
Microscopy and Microanalysis , Volume 7 , Issue S2: Proceedings: Microscopy & Microanalysis 2001, Microscopy Society of America 59th Annual Meeting, Microbeam Analysis Society 35th Annual Meeting, Long Beach, California August 5-9, 2001 , August 2001 , pp. 614 - 615
Copyright
Copyright © Microscopy Society of America 2001

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

1.Jahn, R, Sudhof, TC.Membrane fusion and exocytosis. Annu Rev Biochem. 1999;68:863-911. Review.Google Scholar
2.Owen, DJ, Luzio, JP.Structural insights into clathrin mediated endocytosis. Curr Opin Cell Biol. 2000 Aug;12(4):467-74.CrossRefGoogle ScholarPubMed
3.Sako, Y, Minoghci, S, Yanagida, T.Single-molecule imaging of EGFR signalling on the surface of living cells. Nat Cell Biol. 2000 Mar;2(3): 168-72.CrossRefGoogle ScholarPubMed
4.Ciechanovar, A, Schwartz, AL, Lodish, HF.Sorting and recycling of cell surface receptors and endocytosed ligands: the asialoglycoprotein and transferrin receptors. J Cell Biochem. 1983;23(l-4):107-30.CrossRefGoogle Scholar