Elasticity mapping of living fibroblasts by AFM and immunofluorescence observation of the cytoskeleton

Using the force mapping mode of atomic force microscopy (AFM), we measured spatial distribution of elastic moduli of living mouse fibroblasts (NIH3T3) in a physiological condition. The nuclear portion of the cellular surface is about 10 times softer than the surroundings. Stiffer fibers are confirme...

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Veröffentlicht in:Ultramicroscopy 2000-02, Vol.82 (1), p.253-258
Hauptverfasser: Haga, Hisashi, Sasaki, Shigeo, Kawabata, Kazushige, Ito, Etsuro, Ushiki, Tatsuo, Sambongi, Takashi
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container_issue 1
container_start_page 253
container_title Ultramicroscopy
container_volume 82
creator Haga, Hisashi
Sasaki, Shigeo
Kawabata, Kazushige
Ito, Etsuro
Ushiki, Tatsuo
Sambongi, Takashi
description Using the force mapping mode of atomic force microscopy (AFM), we measured spatial distribution of elastic moduli of living mouse fibroblasts (NIH3T3) in a physiological condition. The nuclear portion of the cellular surface is about 10 times softer than the surroundings. Stiffer fibers are confirmed in the elastic images. In order to investigate origin of the softer nuclear portion and the stiffer fibers, we fixed the identical cells imaged by the AFM, and carried out immunofluorescence observation for three types of cytoskeletal filaments — actin filaments, microtubules, and intermediate filaments, using confocal laser scanning microscopy (CLSM). A comparison between the AFM and the CLSM images revealed that the elasticity of the cells was concerned not only with the distribution of actin network, but also with intermediate filaments, whereas microtubules had no large effect on the measured elasticity.
doi_str_mv 10.1016/S0304-3991(99)00157-6
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subjects 3T3 Cells
Actins - ultrastructure
Animal cell culture
Animals
Atomic force microscopy
Cytoskeleton
Cytoskeleton - ultrastructure
Elastic moduli
Elastic modulus
Elasticity
Fibroblast
Fibroblasts - chemistry
Fibroblasts - ultrastructure
Fluorescence
Fluorescent Antibody Technique
Laser applications
Medical imaging
Mice
Microscopy, Atomic Force - methods
Microscopy, Confocal - methods
title Elasticity mapping of living fibroblasts by AFM and immunofluorescence observation of the cytoskeleton
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