[Near-field microscopy: from the isolated molecule to the living cell]

Abstract

Near field (or scanning probe) microscopy is a recent technology which, owing to the huge amount of publications, is becoming a reference method in molecular and cellular imaging. These microscopies consist in the scanning of the sample, line by line, with a very tiny tip and thus providing informations on its surface down to the nanometer scale. These methods gather scanning tunelling microscopy (STM), which measures a current between the tip and the specimen support, atomic force microscopy (AFM), which measures the repulsive and attractive forces of the tip in contact or very close to the specimen, and scanning near field optical microscopies (SNOM), for which a glass tip allows to catch light signals. Atomic force microscopy, which allows the observation of specimens in air or physiological conditions environments, is presently dominant in biology, in complementarity with the classical optical and electron microscopies, which by the way, have also shown considerable improvements during the last years. The complementarity of these microscopies is due to their very different basic principles, which provide them various possibilities and limits. The biological applications of STM is limited by the need of conducting samples, but the different models of SNOM, often still in development, allow to consider very interesting applications, particularly for detecting very faint and tiny fluorescence signals. Different examples will be given concerning the visualization by AFM of isolated DNA molecules, naked or associated with proteins, the observation of intact or decondensed chromosomes, as well as living cells. One of the originality of AFM is its capacity to observe objects in a wide range of enlargements, with fields from a few hundred of nanometers to several micrometers.