Quantifying Heat Stress Response in C. elegans

Approximately 200 L1 larvae from Psod-3::gfp synchronized animals were seeded on 60 mm plates of NGM media supplemented with 0, 20, 40, 80 or 100 mM glucose and incubated at 20°C until they reached L4 larval stage. At this stage plates were transferred to an air incubator at 31°C for 8 h as indicated [29 ]. After heat shock, animals were anesthetized with 10 mM tetramisole, mounted on agarose pads at 2% and observed on a Nikon Eclipse E600 microscope equipped with an AxioCam MRc camera and Zeiss AxioVision software. Quantification of GFP fluorescence was performed using Image J software as previously indicated [30 (link)].

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Alcántar-Fernández J., Navarro R.E., Salazar-Martínez A.M., Pérez-Andrade M.E, & Miranda-Ríos J. (2018). Caenorhabditis elegans respond to high-glucose diets through a network of stress-responsive transcription factors. PLoS ONE, 13(7), e0199888.

Publication 2018

Eclipse E600 microscope AxioCam MRc camera AxioVision software

Agarose Animals E600 Fluorescence Glucose Heat shock Larval Microscope Tetramisole

Corresponding Organization : Universidad Nacional Autónoma de México

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Variable analysis

independent variables

Glucose concentration (0, 20, 40, 80, or 100 mM)

dependent variables

GFP fluorescence

control variables

L1 larval stage of Psod-3::gfp synchronized animals
NGM media
Incubation temperature at 20°C until L4 larval stage
Heat shock at 31°C for 8 hours
Anesthesia with 10 mM tetramisole
Agarose pads at 2%
Nikon Eclipse E600 microscope with AxioCam MRc camera and Zeiss AxioVision software
Image J software for quantification of GFP fluorescence

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