Analysis of human fibroblasts in vitro-imaging conditions and cytochalasintreatment

The structure of human fibroblasts has been characterised in vitro by atomi c force microscopy (AFM) operated in the imaging or in the force vs. distan ce (F-d) modes. The choice of growth substrate is important to ensure good adhesion. The substrate also affects the imaging conditions for in vitro an alysis of live cells; activated tissue culture dishes are shown to promote conditions that routinely result in good quality images. A qualitative mode l suggests that the activated substrate may act as a preferential scavenger of cellular debris, therefore promoting low adhesion between tip and membr ane and preventing the tip from biofouling. Alternatively, the activated su bstrate may promote a more rigid cell structure, thus resulting in improved imaging. Good imaging conditions provide nondestructive in vitro informati on about cytoskeletal structure and dynamics; thus, treatment with cytochal asin can be monitored in real time for durations of several hours. (C) 1999 Elsevier Science B.V. All rights reserved.