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AIM:To establish a method for optical sections of HepG2human hepatoblastoma cells with confocal laser scanningmicroscope (CLSM) and to study the spatial structure offilamentous actin (F-actin) in HepG2 cells.METHODS:HepG2 cells were stained with FITC-phalloidinthat specifically binds F-actin,with propidium iodide (PI) tothe nucleus,and scanned with a CLSM to generate opticallysectioned images.A series of optical sections takensuccessively at different focal levels in steps of 0.7μm werereconstructed with the CLSM reconstruction program.RESULTS:CLSM images showed that the FITC-stained F-actin was abundant microfilament bundles parallel or nettedthrough the whole cell and its processes.Most F-actinmicrofilaments extended through the cell from one part towardthe other or run through the process.Some microfilamentswere attached to the plasma membrane,or formed astructural bridge connecting to the neighboring cells.CONCLUSION:A method for double labeling HepG2 humanhepatoblastoma cells and CLSM imaging F-actin microfilamentsand nuclei by image thin optical sections and spatial structurewas developed.It provides a very useful way to study thespatial structure of F-actin.
AIM: To establish a method for optical sections of HepG2human hepatoblastoma cells with confocal laser scanning microscope (CLSM) and to study the spatial structure offilamentous actin (F-actin) in HepG2 cells. METHODS: HepG2 cells were stained with FITC-phalloid interface specifically binds F -actin, with propidium iodide (PI) tothe nucleus, and scanned with a CLSM to generate optically blocked images. A series of optical sections takensuccessively at different focal levels in steps of 0.7 μm werereconstructed with the CLSM reconstruction program. RESULTS: CLSM images showed that the FITC-stained F-actin was abundant microfilament bundles parallel or nettedthrough the whole cell and its processes. Host F-actinmicrofilaments extended through the cell from one part toward the other or run through the process. Home microfilamentswere attached to the plasma membrane, or formed astructural 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 structurewas developed. It provides a very useful way to study the spatial structure of F-actin.