Photonic lantern TIRF microscopy for highly efficient, uniform, artifact-free imaging

Area of Science:

• Optical Microscopy
• Biophysics
• Photonics

Background:

• Total internal reflection fluorescence (TIRF) microscopy is crucial for live-cell imaging, but achieving uniform excitation can be challenging.
• Existing TIRF methods often involve complex setups with moving parts, limiting their practicality and robustness.

Purpose of the Study:

• To introduce a novel method for generating uniform, artifact-free TIRF excitation using a photonic lantern.
• To overcome the limitations of conventional TIRF illumination systems.

Main Methods:

• A tapered waveguide photonic lantern with a multimode input and nine few-mode outputs was designed and fabricated.
• The photonic lantern was utilized for simultaneous multi-beam TIRF illumination from nine azimuthal directions.
• The system's performance was characterized, including TIRF penetration depth tuning and light source compatibility.

Main Results:

• The photonic lantern enabled single-shot, uniform, and artifact-free TIRF excitation without non-stationary devices.
• The system demonstrated low-loss light transmission, supporting various light sources and wavelengths.
• High-quality excitation facilitated unbiased imaging across the entire field-of-view.

Conclusions:

• The photonic lantern TIRF (PL-TIRF) technique offers a simple, robust, and advantageous alternative to existing TIRF methods.
• PL-TIRF enables advanced imaging applications, including flat-field super-resolution and shadowless live-cell imaging.