Filamentous-actins in human hepatocarcinoma cells with CLSM

World J Gastroenterol. 2004 Jun 1;10(11):1666-8. doi: 10.3748/wjg.v10.i11.1666.

Abstract

Aim: To establish a method for optical sections of HepG2 human hepatoblastoma cells with confocal laser scanning microscope (CLSM) and to study the spatial structure of filamentous actin (F-actin) in HepG2 cells.

Methods: HepG2 cells were stained with FITC-phalloidin that specifically binds F-actin, with propidium iodide (PI) to the nucleus, and scanned with a CLSM to generate optically sectioned images. A series of optical sections taken successively at different focal levels in steps of 0.7 microm were reconstructed with the CLSM reconstruction program.

Results: CLSM images showed that the FITC-stained F-actin was abundant microfilament bundles parallel or netted through the whole cell and its processes. Most F-actin microfilaments extended through the cell from one part toward the other or run through the process. Some microfilaments were attached to the plasma membrane, or formed a structural bridge connecting to the neighboring cells.

Conclusion: A method for double labeling HepG2 human hepatoblastoma cells and CLSM imaging F-actin microfilaments and nuclei by image thin optical sections and spatial structure was developed. It provides a very useful way to study the spatial structure of F-actin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actin Cytoskeleton / metabolism
  • Actin Cytoskeleton / ultrastructure*
  • Actins / metabolism*
  • Actins / ultrastructure
  • Carcinoma, Hepatocellular*
  • Cell Line, Tumor / metabolism
  • Cell Line, Tumor / ultrastructure
  • Fluorescent Dyes
  • Humans
  • Imaging, Three-Dimensional
  • Liver Neoplasms*
  • Microscopy, Confocal / methods*
  • Phalloidine / analogs & derivatives*

Substances

  • Actins
  • Fluorescent Dyes
  • fluorescein isothiocyanate-phalloidin
  • Phalloidine