Fluorescence microscopy, utilizing fluorescence labeling, has the capability to observe intercellular changes which transmitted and reflected light microscopy techniques cannot resolve. However, the parts without fluorescence labeling are not imaged. Hence, the processes simultaneously happen in these parts cannot be revealed. Meanwhile, fluorescence imaging is 2D imaging where information in the depth is missing. Therefore the information in labeling parts is also not complete.
On the other hand, quantitative phase imaging is capable to image cells in 3D in real time through phase calculation. However, its resolution is limited by the optical diffraction and cannot observe intercellular changes below 200 nanometers.
In this work, fluorescence imaging and quantitative phase imaging are combined to build a multimodal imaging system. Such system has the capability to simultaneously observe the detailed intercellular phenomenon and 3D cell morphology. In this study the proposed multimodal imaging system is used to observe the cell behavior in the cell apoptosis. The aim is to highlight the limitations of fluorescence microscopy and to point out the advantages of multimodal quantitative phase and fluorescence imaging. The proposed multimodal quantitative phase imaging could be further applied in cell related biomedical research, such as tumor.
Digital holography (DH) is a 3D imaging technique with a theoretical axial accuracy of better than 1-2 nanome-ters. However, practically, the axial error has been quoted to be tens of nanometers which is much larger than the theoretical value. Previous studies of the axial error mainly focused on the phase error introduced by lens. However, it is found that CCD aperture size is also an important contributors to axial error by our group. It is necessary to investigate the reduction approach of such axial error. The most possible connection between the limited CCD aperture size and the axial error is the diffraction effect. Window functions once have been applied to digital holograms for diffraction suppression and improve the lateral resolution of the intensity image. How-ever, their impacts on phase image and the associated axial dimension measurement are still unknown. In this paper, window functions are applied to digital holograms for phase/axial error reduction. Both simulation and experiment are performed. Moreover, the relation between axial error and window functions is also illustrated by the mathematical formulas derived in the theory. And all the results validate that the window functions can reduce the axial error of digital holography.