PMID- 24692492
OWN - NLM
STAT- MEDLINE
DCOM- 20141114
LR  - 20140402
IS  - 1559-6095 (Electronic)
IS  - 1559-6095 (Linking)
VI  - 2014
IP  - 4
DP  - 2014 Apr 1
TI  - Fluorescent visualization of germline apoptosis in living Caenorhabditis elegans.
PG  - 420-7
LID - 10.1101/pdb.prot080226 [doi]
AB  - Visualization of apoptosis using fluorescent tools is quite straightforward in 
      living Caenorhabditis elegans. Several transgenic lines are available that mark 
      dying cells with fluorescent proteins, making it possible to quantify kinetics at 
      various stages of the apoptotic process. Proteins required for the engulfment of 
      cell corpses are particularly useful for detecting early apoptotic stages using 
      this approach. For example, expression of the engulfment protein CED-1 fused to 
      green fluorescent protein (CED-1::GFP) creates a halo around cells during early 
      apoptosis, before their refractile morphology can be detected by differential 
      interference contrast (DIC) optics. In addition, vital dyes such as acridine 
      orange (AO) and SYTO-12 are selectively retained in apoptotic cells and can be 
      used to visualize apoptosis in the germlines of living animals. It is also 
      possible to use vital dyes in combination with transgenic strains expressing 
      fluorescent markers of cell corpses to examine, in detail, multiple stages of 
      apoptosis in vivo. Because of the high optical contrast of fluorescent reagents, 
      apoptosis can be visualized even at low magnification, facilitating the use of 
      screening platforms to identify apoptosis regulators. This protocol describes 
      multiple uses of fluorescent reagents for visualization of germline apoptosis in 
      living C. elegans, including AO staining, time-course studies using fluorescent 
      proteins, and low-throughput screening of cell death genes using RNA interference 
      (RNAi).
FAU - Lant, Benjamin
AU  - Lant B
AD  - Developmental and Stem Cell Biology Program, The Hospital for Sick Children, 
      Toronto, Ontario M5G 1L7, Canada;
FAU - Derry, W Brent
AU  - Derry WB
LA  - eng
PT  - Journal Article
DEP - 20140401
PL  - United States
TA  - Cold Spring Harb Protoc
JT  - Cold Spring Harbor protocols
JID - 101524530
RN  - 0 (Fluorescent Dyes)
RN  - 0 (Luminescent Proteins)
SB  - IM
MH  - Animals
MH  - *Apoptosis
MH  - Caenorhabditis elegans/*physiology
MH  - Fluorescent Dyes/*analysis
MH  - Germ Cells/*physiology
MH  - Luminescent Proteins/*analysis
MH  - Optical Imaging/*methods
MH  - Staining and Labeling/*methods
EDAT- 2014/04/03 06:00
MHDA- 2014/11/15 06:00
CRDT- 2014/04/03 06:00
PHST- 2014/04/03 06:00 [entrez]
PHST- 2014/04/03 06:00 [pubmed]
PHST- 2014/11/15 06:00 [medline]
AID - 2014/4/pdb.prot080226 [pii]
AID - 10.1101/pdb.prot080226 [doi]
PST - epublish
SO  - Cold Spring Harb Protoc. 2014 Apr 1;2014(4):420-7. doi: 10.1101/pdb.prot080226.