Volumetric live-cell autofluorescence imaging using Fourier light-field microscopy

Area of Science:

• Cell Biology
• Microscopy
• Biophysics

Background:

• Live-cell imaging is crucial for understanding cellular dynamics.
• Traditional methods often require exogenous labels or cause significant photodamage.
• Visualizing native cellular autofluorescence in 3D remains challenging.

Purpose of the Study:

• To develop a rapid, volumetric live-cell imaging technique.
• To visualize autofluorescent sub-cellular structures and their dynamics in 3D.
• To enable multicolor imaging of native and labeled organelles.

Main Methods:

• High-resolution Fourier light-field microscopy.
• Volumetric live-cell imaging.
• Multicolor imaging of autofluorescent and fluorescently-labeled organelles.

Main Results:

• Successfully captured lysosomal autofluorescence in fibroblasts and HeLa cells.
• Simultaneously imaged lysosomal autofluorescence and fluorescently-labeled lysosomes/mitochondria.
• Quantified interactions between lysosomes and mitochondria in live cells.

Conclusions:

• The developed technique enables rapid 3D live-cell imaging with reduced photodamage.
• It eliminates the need for exogenous labels for visualizing certain cellular structures.
• This method supports future studies of native cellular states and functions in 3D.