Fast wide-field light sheet electro-optic FLIM

Area of Science:

• Biophotonics
• Microscopy
• Molecular Imaging

Background:

• Fluorescence Lifetime Microscopy (FLIM) provides functional and molecular information.
• Traditional FLIM methods can be slow for volumetric imaging.
• Selective Plane Illumination Microscopy (SPIM) offers optical sectioning for 3D imaging.

Purpose of the Study:

• To demonstrate volumetric FLIM using electro-optic gating in a SPIM setup.
• To enable faster 3D FLIM acquisition.
• To image biological samples with high spatial and temporal resolution.

Main Methods:

• Implemented electro-optic FLIM (EO-FLIM) with a Pockels cell.
• Integrated EO-FLIM gating with a selective plane illumination microscope (SPIM).
• Achieved light sheet electro-optic FLIM (LS-EO-FLIM) with an 80 MHz gating frequency and 800 µm field of view.

Main Results:

• Demonstrated successful volumetric imaging of fluorescent bead mixtures.
• Acquired 3D FLIM data from live Arabidopsis thaliana root samples.
• Utilized both genetically encoded fluorescent proteins and endogenous autofluorescence for imaging.

Conclusions:

• LS-EO-FLIM is a viable technique for rapid volumetric fluorescence lifetime imaging.
• The method allows for imaging of diverse biological samples.
• This technique advances 3D functional imaging in live biological systems.