Lipofuscin Accumulation and Pumping Rate Analysis in Aging Worms

About 10 worms of each condition were analyzed at the early stage (from day 1 to day 4) and during aging (from day 9 to day 11) of adulthood. Pumping rate was measured under Zeiss Axiovert 25 microscope by counting the number of grinder contractions of 10 animals for each treatment, during a period of 30 s, as described in (Guantario et al., 2018 (link)). To analyze the auto-fluorescence of lipofuscin granules, 10 worms per condition at different stages (day 2, 3, 5 and 11) were washed three times with M9 buffer and observed by Zeiss Axiovert 25 microscope under DAPI filter. Images were taken at the time of exposure of 0.2 s and scale bars were inserted by Zeiss ZEN Microscopy Software 2011. Fluorescence was analyzed using ImageJ software. Aging analyses were performed in triplicate.

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Schifano E., Conta G., Preziosi A., Ferrante C., Batignani G., Mancini P., Tomassini A., Sciubba F., Scopigno T., Uccelletti D, & Miccheli A. (2022). 2-hydroxyisobutyric acid (2-HIBA) modulates ageing and fat deposition in Caenorhabditis elegans. Frontiers in Molecular Biosciences, 9, 986022.

Publication 2022

Zeiss Axiovert 25 microscope ZEN Microscopy Software 2011

Animals Buffer Dapi Fluorescence Granules Lipofuscin Microscopy Worms

Corresponding Organization : Sapienza University of Rome

Other organizations : Italian Institute of Technology

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Variable analysis

independent variables

Conditions (not explicitly mentioned)

dependent variables

Pumping rate (measured as number of grinder contractions in 30 seconds)
Auto-fluorescence of lipofuscin granules

control variables

Time points: early stage (day 1 to day 4) and aging (day 9 to day 11) of adulthood
Number of worms analyzed: about 10 worms of each condition

controls

No positive or negative controls were explicitly mentioned in the protocol.

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