Fluorescence Lifetime Imaging of PolyQ Protein Aggregation in Caenorhabditis elegans Neurons

Begin with anesthetized, age-matched control and chaperone-deficient Caenorhabditis elegans worms.

Both groups express fluorophore-tagged polyQ proteins in their neurons that are prone to misfolding.

In control worms, chaperone proteins limit misfolded polyQ aggregation, whereas, in chaperone-deficient worms, misfolded polyQ readily forms aggregates.

Place the control slide on a confocal microscope stage for Fluorescence Lifetime Imaging Microscopy (FLIM).

Illuminate the worms using a pulsed laser.

The laser excites the fluorophore, which emits a photon as it returns to the ground state.

FLIM measures fluorescence lifetime—the duration the fluorophore remains in its excited state.

In chaperone-deficient worms, polyQ aggregation clusters fluorophores, promoting energy transfer between adjacent fluorophores instead of photon emission, thereby reducing fluorescence lifetime.

The FLIM software processes these lifetimes to generate color-coded maps, enabling aggregation visualization.

A reduced fluorescence lifetime in chaperone-deficient worms, compared to controls, indicates increased polyQ aggregation.

Make sure the nematodes are completely immobile. Open the FLIM acquisition software.