Three-dimensional microscopic imaging through scattering media based on in-line phase-shift digital holography

Area of Science:

• Optics and Photonics
• Biomedical Imaging
• Microscopy

Background:

• Scattering media obscure direct imaging of internal structures.
• Quantitative phase imaging offers label-free contrast but is sensitive to scattering.
• Digital holography provides 3D information but struggles with scattering effects.

Purpose of the Study:

• To develop a method for microscopic 3D imaging and phase quantification of objects obscured by scattering media.
• To demonstrate high spatial resolution and accuracy in phase imaging through scattering.
• To validate the technique for imaging through complex biological tissues.

Main Methods:

• In-line phase-shift digital holography was employed.
• Microscopic imaging setup was configured for phase retrieval.
• Scattering media, including a scatter plate and rat skin, were used for testing.

Main Results:

• Achieved a spatial resolution of 1.81 µm for objects behind a scatter plate.
• Demonstrated highly accurate quantitative phase imaging through scattering.
• Successfully performed 3D imaging with a depth difference of 1.32 mm.
• Obtained near-theoretical spatial resolution when imaging through rat skin.

Conclusions:

• In-line phase-shift digital holography enables effective microscopic 3D imaging and phase quantification through scattering media.
• The method offers high resolution and accuracy, suitable for complex biological samples.
• This technique advances label-free imaging capabilities for obscured structures.