Photobleaching regions of living cells to monitor membrane traffic

Erik Lee Snapp, Patrick Lajoie

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Eukaryotic cells are composed of an intricate system of internal membranes that are organized into different compartments-including the endoplasmic reticulum (ER), the nuclear envelope, the Golgi complex (GC), lysosomes, endosomes, caveolae, mitochondria, and peroxisomes-that perform specialized tasks within the cell. The localization and dynamics of intracellular compartments are now being studied in living cells because of the availability of green fluorescent protein (GFP)- fusion proteins and recent advances in fluorescent microscope imaging systems. This protocol outlines two methods for photobleaching living cells to monitor membrane traffic. The first method involves selective photobleaching using a confocal laser-scanning microscope (CLSM) that can bleach discrete selected regions of interest. As outlined in the second method, photobleaching can also be performed with older CLSMs that lack the capacity for selective photobleaching. In this case, photobleaching is accomplished by zooming into a small region of the cell and scanning with full laser power.

Original languageEnglish (US)
Pages (from-to)1366-1367
Number of pages2
JournalCold Spring Harbor Protocols
Volume6
Issue number11
DOIs
StatePublished - Nov 2011

Fingerprint

Photobleaching
Cells
Membranes
Lasers
Microscopes
Scanning
Caveolae
Mitochondria
Peroxisomes
Endosomes
Nuclear Envelope
Golgi Apparatus
Eukaryotic Cells
Green Fluorescent Proteins
Lysosomes
Endoplasmic Reticulum
Imaging systems
Fusion reactions
Availability
Proteins

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Photobleaching regions of living cells to monitor membrane traffic. / Snapp, Erik Lee; Lajoie, Patrick.

In: Cold Spring Harbor Protocols, Vol. 6, No. 11, 11.2011, p. 1366-1367.

Research output: Contribution to journalArticle

Snapp, Erik Lee ; Lajoie, Patrick. / Photobleaching regions of living cells to monitor membrane traffic. In: Cold Spring Harbor Protocols. 2011 ; Vol. 6, No. 11. pp. 1366-1367.
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