Speckle-structured illumination for 3D phase and fluorescence computational microscopy.

Area of Science:

• Biomedical Engineering
• Optical Microscopy
• Computational Imaging

Background:

• High-content biological microscopy requires high-resolution imaging over large fields-of-view.
• Computational imaging approaches are crucial for achieving super-resolution in microscopy.
• Previous work demonstrated 2D multimodal high-content microscopy using speckle illumination structured illumination microscopy (SIM).

Purpose of the Study:

• To extend the 2D multimodal high-content microscopy method to 3D.
• To develop algorithms for joint retrieval of 3D super-resolved fluorescent and complex-field distributions.
• To achieve 3D imaging beyond the diffraction limit.

Main Methods:

• Leveraging speckle illumination as a 3D structured pattern.
• Employing both coherent and incoherent imaging models.
• Developing algorithms for joint retrieval of 3D sample distributions.

Main Results:

• Demonstrated 3D super-resolution beyond the physical diffraction limit.
• Achieved 3D multimodal imaging with $$ m³ resolution.
• Successfully imaged a volume of $$ m³.

Conclusions:

• The developed 3D method effectively extends super-resolution microscopy capabilities.
• Speckle illumination provides a viable 3D structured pattern for advanced microscopy.
• This technique enables high-resolution 3D imaging for biological applications.