Exploration of Deformation of F-Actin during Macropinocytosis by Confocal Microscopy and 3D-Structured Illumination Microscopy

Abstract

Since their invention, confocal microscopy and super-resolution microscopy have become important choices in cell biology research. Macropinocytosis is a critical form of endocytosis. Deformation of the cell membrane is thought to be closely related to the movement of F-actin during macropinocytosis. However, it is still unclear how the morphology of F-actin and the membrane change during this process. In this study, confocal microscopy was utilized for macroscopic time-series imaging of the cell membranes and F-actin in cells induced by phorbol 12-myristate 13-acetate (PMA). Super-resolution structured illumination microscopy (SIM), which can overcome the diffraction limit, was used to demonstrate the morphological characteristics of F-actin filaments. Benefiting from the advantages of SIM in terms of resolution and 3D imaging, we speculated on the regular pattern of the deformation of F-actin during macropinocytosis. The detailed visualization of structures also helped to validate the speculation regarding the role of F-actin filaments in macropinocytosis in previous studies. The results obtained in this study will provide a better understanding of the mechanisms underlying macropinocytosis and endocytosis.

Keywords

• F-actin
• macropinocytosis
• 3D-SIM
• confocal microscopy
• morphological study