Extended Dual-Focus Microscopy for Ratiometric-Based 3D Movement Tracking

Abstract

Imaging the three-dimensional movement of small organelles in living cells can provide key information for the dynamics of drug delivery and virus transmission in biomedical disciplines. To stably monitor such intracellular motion using microscope, long depth of field along optical axis and accurate three-dimensional tracking are simultaneously required. In the present work, we suggest an extended dual-focus optics microscopy system by combining a bifocal plane imaging scheme and objective lens oscillation, which enables accurate localization for a long axial range. The proposed system exploits high-resolution functionality by concatenating partial calibration result acquired each axial imaging level, maintaining the practical advantages of ratiometric method.

Visualization Method for the Cell-Level Vesicle Transport Using Optical Flow and a Diverging Colormap

“…In addition, because the proposed method assumes the vesicles moves within the depth of the image plane, three-dimensional movements of some vesicles that can be shown as appearing or disappearing in the image plane might be wrongly detected. In this case, multi-plane imaging [ 47 ] is expected to help exclude such spurious detection, by comparing the images acquired from vertically separated nearest image planes.…”

“…In addition, because the proposed method assumes the vesicles moves within the depth of the image plane, three-dimensional movements of some vesicles that can be shown as appearing or disappearing in the image plane might be wrongly detected. In this case, multi-plane imaging [47] is expected to help exclude such spurious detection, by comparing the images acquired from vertically separated nearest image planes. One of the major advantages of adopting the suggested methods is that various cell experiments conducted to examine the features of biological processes including endocytosis can be analyzed visually.…”