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 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.
Haga
H
H
Division of Physics, Graduate School of Science, Hokkaido University, Sapporo, Japan. haga@skws.sci.hokudai.ac.jp
Sasaki
S
S
Kawabata
K
K
Ito
E
E
Ushiki
T
T
Sambongi
T
T
D016475:3T3 Cells; D000199:Actins / Q000648:ultrastructure; D000818:Animals; D003599:Cytoskeleton / Q000648:ultrastructure; D004548:Elasticity; D005347:Fibroblasts / Q000737:chemistry / Q000648:ultrastructure*; D005455:Fluorescent Antibody Technique; D051379:Mice; D018625:Microscopy, Atomic Force / Q000379:methods; D018613:Microscopy, Confocal / Q000379:methods
D000199:Actins