Chip-based wide field-of-view total internal reflection fluorescence microscopy.

Area of Science:

• Optics and Photonics
• Biomedical Imaging
• Materials Science

Background:

• Conventional total internal reflection fluorescence (TIRF) microscopy relies on high numerical aperture objectives or prisms for evanescent wave generation.
• These methods necessitate precise optical alignment, increasing operational complexity and limiting field-of-view.
• Existing techniques struggle with bulky components and intricate setup procedures.

Purpose of the Study:

• To introduce a planar photonic chip as a simplified alternative for evanescent wave generation in TIRF microscopy.
• To overcome the limitations of conventional TIRF objectives and prisms.
• To enhance ease of use and expand imaging capabilities in TIRF microscopy.

Main Methods:

• A planar photonic chip, comprising a dielectric multilayer and a scattering layer, was designed and fabricated.
• The chip was utilized to excite uniform evanescent waves under uncollimated light incidence.
• The chip's performance was evaluated in a modified TIRF microscopy setup.

Main Results:

• The photonic chip successfully generated uniform evanescent waves, enabling TIRF illumination.
• The proposed chip eliminated the need for precise optical alignment, simplifying the microscopy setup.
• Shadowless illumination and a large field-of-view (FOV) were achieved due to the chip's design and separated light paths.

Conclusions:

• The planar photonic chip offers a compact and user-friendly solution for TIRF microscopy.
• This approach significantly reduces operational complexity and enhances imaging performance.
• The technology holds potential for broader adoption of advanced fluorescence microscopy techniques.