Filamentous-actins in human hepatocarcinoma cells with CLSM.
OBJECTIVE
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.


CONCLUSIONS
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.
Huo
Xia
X
Central Laboratory, Shantou University Medical College, Shantou 515031, Guangdong Province, China. xhuo@stu.edu.cn
Xu
Xi-Jin
XJ
Chen
Yao-Wen
YW
Yang
Hai-Wei
HW
Piao
Zhong-Xian
ZX
D008841:Actin Cytoskeleton / Q000378:metabolism / Q000648:ultrastructure*; D000199:Actins / Q000378:metabolism* / Q000648:ultrastructure; D006528:Carcinoma, Hepatocellular; D045744:Cell Line, Tumor / Q000378:metabolism / Q000648:ultrastructure; D005456:Fluorescent Dyes; D006801:Humans; D021621:Imaging, Three-Dimensional; D008113:Liver Neoplasms; D018613:Microscopy, Confocal / Q000379:methods*; D010590:Phalloidine / Q000031:analogs & derivatives*
D000199:Actins; D005456:Fluorescent Dyes; C037191:fluorescein isothiocyanate-phalloidin; D010590:Phalloidine