Quantitative phase microscopy for cellular dynamics based on transport of intensity equation

Abstract

We demonstrate a simple method for quantitative phase imaging of tiny transparent objects such as living cells based on the transport of intensity equation. The experiments are performed using an inverted bright field microscope upgraded with a flipping imaging module, which enables to simultaneously create two laterally separated images with unequal defocus distances. This add-on module does not include any lenses or gratings and is cost-effective and easy-to-alignment. The validity of this method is confirmed by the measurement of microlens array and human osteoblastic cells in culture, indicating its potential in the applications of dynamically measuring living cells and other transparent specimens in a quantitative, non-invasive and label-free manner.

Real-Time Reconstruction of the Complex Field of Phase Objects Based on Off-Axis Interferometry

“…Both non-interference and interference methods have been widely used in quantitative phase imaging of biological samples. For example, the microscopy based on intensity transfer equations can realize phase imaging of biological samples through a series of numerical operations [3][4][5][6]. However, it is limited by the complexity of the calculation process and the long time required.…”

Measurements on ATP induced cellular fluctuations using real-time dual view transport of intensity phase microscopy

“…Besides, based on the field of view (FoV) division, Waller et al [47], Yang et al [48] and Yu et al [49] proposed single-shot transport of intensity phase microscopy using volume holography, grating and spatial light modulator, respectively. Li et al [50] and Zuo et al [51] also designed mirror-prism based single-shot transport of intensity phase microscopy. Using these methods, multi-focal intensities can be simultaneously collected at a single imaging plane.…”

Quantitative and Dynamic Phase Imaging of Biological Cells by the Use of the Digital Holographic Microscopy Based on a Beam Displacer Unit

“…However, its limitation is the difficulty to achieve quantitative description of the specimens' morphology, especially for biological specimens with low-contrast and blurred boundaries. Transport of intensity equation combined with commercial microscope or the lens-free on-chip microscope is introduced to implement phase imaging of biological specimens [1]- [5]. This technique can achieve phase imaging of pathology slide by a series of digital processing steps to retrieve the original object information.…”