Target-surface multiplexed quantitative dynamic phase microscopic imaging based on the transport-of-intensity...

Area of Science:

• Optics and Photonics
• Biomedical Imaging
• Digital Microscopy

Background:

• Traditional phase imaging relies on mechanical scanning or interferometry, limiting speed and complexity.
• Microscopic phase digital imaging using the Transport of Intensity Equation (TIE) offers an alternative but often requires specialized setups.
• There is a need for simpler, faster, and non-interferometric methods for quantitative phase imaging.

Purpose of the Study:

• To develop a single exposure target-surface multiplexed phase reconstruction (SETMPR) structure based on TIE.
• To achieve dynamic, non-interferometric, quantitative refractive index distribution measurement in a single shot.
• To demonstrate the application of SETMPR in both static optical and dynamic biological sample imaging.

Main Methods:

• Implemented a SETMPR structure by integrating a conventional bright-field inverted microscope with a specialized wavefront-shaping transmission element.
• Utilized the Transport of Intensity Equation (TIE) for phase reconstruction from intensity measurements.
• Validated quantitative accuracy by comparing measurements of a microlens array and grating with a standard instrument.
• Performed in situ static and long-term dynamic imaging of HT22 cells, incorporating machine learning for automatic image segmentation.

Main Results:

• The SETMPR structure is easy to construct and enables dynamic, non-interferometric quantitative phase imaging.
• Measurement accuracy was validated against standard instruments for optical samples.
• Successful quantitative imaging of static and dynamic biological samples (HT22 cells) was achieved with frame-rate-limited speed.
• Machine learning facilitated automatic image segmentation for cellular imaging.

Conclusions:

• The SETMPR method provides a practical and efficient approach for quantitative phase imaging.
• This technique significantly advances dynamic observation of cellular processes in biomedical fields.
• The system's simplicity and speed open new avenues for high-throughput biological imaging and optical metrology.