PMID- 36533827
OWN - NLM
STAT- MEDLINE
DCOM- 20221220
LR  - 20231203
IS  - 1940-087X (Electronic)
IS  - 1940-087X (Linking)
IP  - 190
DP  - 2022 Dec 2
TI  - Long-Term Culture of Individual Caenorhabditis elegans on Solid Media for 
      Longitudinal Fluorescence Monitoring and Aversive Interventions.
LID - 10.3791/64682 [doi]
AB  - Caenorhabditis elegans are widely used to study aging biology. The standard 
      practice in C. elegans aging studies is to culture groups of worms on solid 
      nematode growth media (NGM), allowing the efficient collection of 
      population-level data for survival and other physiological phenotypes, and 
      periodic sampling of subpopulations for fluorescent biomarker quantification. 
      Limitations to this approach are the inability to (1) follow individual worms 
      over time to develop age trajectories for phenotypes of interest and (2) monitor 
      fluorescent biomarkers directly in the context of the culture environment. 
      Alternative culture approaches use liquid culture or microfluidics to monitor 
      individual animals over time, in some cases including fluorescence 
      quantification, with the tradeoff that the culture environment is contextually 
      distinct from solid NGM. The WorMotel is a previously described microfabricated 
      multi-well device for culturing isolated worms on solid NGM. Each worm is 
      maintained in a well containing solid NGM surrounded by a moat filled with copper 
      sulfate, a contact repellent for C. elegans, allowing longitudinal monitoring of 
      individual animals. We find copper sulfate insufficient to prevent worms from 
      fleeing when subjected to aversive interventions common in aging research, 
      including dietary restriction, pathogenic bacteria, and chemical agents that 
      induce cellular stress. The multi-well devices are also molded from 
      polydimethylsiloxane, which produces high background artifacts in fluorescence 
      imaging. This protocol describes a new approach for culturing isolated roundworms 
      on solid NGM using commercially available polystyrene microtrays, originally 
      designed for human leukocyte antigen (HLA) typing, allowing the measurement of 
      survival, physiological phenotypes, and fluorescence across the lifespan. A 
      palmitic acid barrier prevents worms from fleeing, even in the presence of 
      aversive conditions. Each plate can culture up to 96 animals and easily adapts to 
      a variety of conditions, including dietary restriction, RNAi, and chemical 
      additives, and is compatible with automated systems for collecting lifespan and 
      activity data.
FAU - Espejo, Luis
AU  - Espejo L
AD  - Department of Molecular & Cellular Biology, University of Arizona, Tucson.
FAU - Hull, Bradford
AU  - Hull B
AD  - Department of Molecular & Cellular Biology, University of Arizona, Tucson.
FAU - Chang, Leah M
AU  - Chang LM
AD  - Department of Molecular & Cellular Biology, University of Arizona, Tucson.
FAU - DeNicola, Destiny
AU  - DeNicola D
AD  - Department of Molecular & Cellular Biology, University of Arizona, Tucson.
FAU - Freitas, Samuel
AU  - Freitas S
AD  - Department of Molecular & Cellular Biology, University of Arizona, Tucson.
FAU - Silbar, Vanessa
AU  - Silbar V
AD  - Department of Molecular & Cellular Biology, University of Arizona, Tucson.
FAU - Haskins, Anne
AU  - Haskins A
AD  - Department of Molecular & Cellular Biology, University of Arizona, Tucson.
FAU - Turner, Emily A
AU  - Turner EA
AD  - Department of Molecular & Cellular Biology, University of Arizona, Tucson.
FAU - Sutphin, George L
AU  - Sutphin GL
AD  - Department of Molecular & Cellular Biology, University of Arizona, Tucson; 
      sutphin@arizona.edu.
LA  - eng
GR  - R35 GM133588/GM/NIGMS NIH HHS/United States
GR  - T32 GM008659/GM/NIGMS NIH HHS/United States
GR  - T32 GM139779/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PT  - Video-Audio Media
DEP - 20221202
PL  - United States
TA  - J Vis Exp
JT  - Journal of visualized experiments : JoVE
JID - 101313252
RN  - LRX7AJ16DT (Copper Sulfate)
RN  - 0 (Culture Media)
RN  - 0 (Biomarkers)
SB  - IM
MH  - Animals
MH  - Humans
MH  - *Caenorhabditis elegans/genetics
MH  - *Copper Sulfate
MH  - Longevity/genetics
MH  - Aging/physiology
MH  - Phenotype
MH  - Culture Media
MH  - Biomarkers
PMC - PMC10368144
MID - NIHMS1909497
EDAT- 2022/12/20 06:00
MHDA- 2022/12/21 06:00
PMCR- 2023/12/02
CRDT- 2022/12/19 08:03
PHST- 2022/12/19 08:03 [entrez]
PHST- 2022/12/20 06:00 [pubmed]
PHST- 2022/12/21 06:00 [medline]
PHST- 2023/12/02 00:00 [pmc-release]
AID - 10.3791/64682 [doi]
PST - epublish
SO  - J Vis Exp. 2022 Dec 2;(190):10.3791/64682. doi: 10.3791/64682.