Experimental setup combining digital holographic microscopy (DHM) and fluorescence imaging to study gold nanoparticle mediated laser manipulation

Abstract

Our research combines Digital Holographic Microscopy (DHM) and ˛uorescence microscopy to study the basic mechanisms of gold nanoparticle mediated laser manipulation. Herein we describe the technical aspects of the setup and holographic image reconstruction. Furthermore, results pertaining to cell volume change and calcium response of cells in laser manipulation will be presented and discussed. For the reconstruction of phase images from fringe image data, a phase unwrapping algorithm is presented that shows great potential to cope with the vast amount of data that was captured. This algorithm is a hybrid between a tile unwrapping technique and a path following unwrapper. It combines the robustness of a path following algorithm and a parallelizable tile unwrapping preprocessing step. The experimental setup enables simultaneous acquisition of ˛uorescence and phase images. For cell manipulation, a picosecond laser was coupled into the setup and weakly focused on cells incubated with gold nanoparticles. To study the cell volume change in the ˝rst minute, phase images were captured with a frame rate of 33 fps. Fluorescence images yielded the calcium signal of the cells as well as the dynamics of the F-actin cytoskeleton after irradiation. The setup is suitable to study fast changes in biophysical and morphological para