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

Abstract

Microscopic phase digital imaging based on the transport of intensity equation, known as TIE, is widely used in optical measurement and biomedical imaging since it can dispense with the dependence of traditional phase imaging systems on mechanical rotational scanning and interferometry devices. In this work, we provide a single exposure target-surface multiplexed phase reconstruction (SETMPR) structure based on TIE, which is remarkably easy to construct since it directly combines a conventional bright-field inverted microscope with a special image plane transmission structure that is capable of wavefront shaping and amplification. In practice, the SETMPR is able to achieve dynamic, non-interferometric, quantitative refractive index distribution of both static optical samples and dynamic biological samples in only one shot, meaning that the only limitation of measuring frequency is the frame rate. By comparing the measurement results of a microlens array and a grating with a standard instrument, the quantitative measurement capability and accuracy are demonstrated. Subsequently, both in situ static and long-term dynamic quantitative imaging of HT22 cells were performed, while automatic image segmentation was completed by introducing machine learning methods, which verified the application prospect of this work in dynamic observation of cellular in the biomedical field.