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 language | English (US) |
|---|---|
| Pages (from-to) | 1366-1367 |
| Number of pages | 2 |
| Journal | Cold Spring Harbor Protocols |
| Volume | 6 |
| Issue number | 11 |
| DOIs | |
| State | Published - Nov 2011 |
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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 journal › Article
}
TY - JOUR
T1 - Photobleaching regions of living cells to monitor membrane traffic
AU - Snapp, Erik Lee
AU - Lajoie, Patrick
PY - 2011/11
Y1 - 2011/11
N2 - 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.
AB - 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.
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U2 - 10.1101/pdb.prot066563
DO - 10.1101/pdb.prot066563
M3 - Article
C2 - 22046038
AN - SCOPUS:80455127029
VL - 6
SP - 1366
EP - 1367
JO - Cold Spring Harbor Protocols
JF - Cold Spring Harbor Protocols
SN - 1559-6095
IS - 11
ER -