We describe a technique for studying living cells with the atomic forc
e microscope (AFM) in tapping mode using a thermostated, controlled-en
vironment culture system. We also describe the integration of the AFM
with bright field, epifluorescence and surface interference microscopy
, achieving the highest level of integration for the AFM thus far desc
ribed. We succeeded in the continuous, longterm imaging of relatively
flat but very fragile cytoplasmic regions of COS cells at a lateral re
solution of about 70 nm and a vertical resolution of about 3 nm. In ad
dition, we demonstrate the applicability of our technology for continu
ous force volume imaging of cultured vertebrate cells. The hybrid inst
rument we describe can be used to collect simultaneously a diverse var
iety of physical, chemical and morphological data on living vertebrate
cells, The integration of light microscopy with AFM and steady-state
culture methods for vertebrate cells represents a new approach for stu
dies in cell biology and physiology.