Temperature-dependent imaging of living cells by AFM.
Characterization of lateral organization of plasma membranes is a prerequisite to the understanding of membrane structure-function relationships in living cells. Lipid-lipid and lipid-protein interactions are responsible for the existence of various membrane microdomains involved in cell signalization and in numerous pathologies. Developing approaches for characterizing microdomains associate identification tools like recognition imaging with high-resolution topographical imaging. Membrane properties are markedly dependent on temperature. However, mesoscopic scale topographical information of cell surface in a temperature range covering most of cell biology experimentation is still lacking. In this work we have examined the possibility of imaging the temperature-dependent behavior of eukaryotic cells by atomic force microscopy (AFM). Our results establish that the surface of living CV1 kidney cells can be imaged by AFM, between 5 and 37 degrees C, both in contact and tapping modes. These first temperature-dependent data show that large cell structures appeared essentially stable at a microscopic scale. On the other hand, as shown by contact mode AFM, the surface was highly dynamic at a mesoscopic scale, with marked changes in apparent topography, friction, and deflection signals. When keeping the scanning conditions constant, a progressive loss in the image contrast was however observed, using tapping mode, on decreasing the temperature.
Institut National de la Santé et de la Recherche Médicale, Unité 554, Montpellier, France.
Cedric
C
Espenel
Marie-Cécile
MC
Giocondi
Bastien
B
Seantier
Patrice
P
Dosset
Pierre-Emmanuel
PE
Milhiet
Christian
C
Le Grimellec
D000818:Animals; D002460:Cell Line; D002522:Chlorocebus aethiops; D007668:Kidney / Q000166:cytology* / Q000648:ultrastructure*; D018625:Microscopy, Atomic Force / Q000379:methods*; D013499:Surface Properties; D013696:Temperature