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Abstract
Newborn rat oligodendrocyte cultures were investigated by scanning near-field optical
microscope (SNOM), a versatile new tool able to map cell membranes in 3D and simultaneously
obtain images of the cytoplasm. Topography, error, transmission and reflection signals
were acquired to describe cell morphology with nanometer-scale resolution. Oligodendrocytes
were studied as a model because their extensive membrane processes (typical of their
physiological role in myelination) made them particularly suitable to test the sensitivity
of the new method. Furthermore, we combined a classical histochemical method with
SNOM, to identify specific intracellular proteins at high definition. In particular,
with this technique, cytoskeleton elements of oligodendrocytes, such as microtubules,
were observed with tubulin antibodies. Images obtained with SNOM were also compared
with those from conventional scanning electron microscopy (SEM) and optical microscopy.
Our results showed that SNOM allowed to observe cell nanostructures otherwise undetectable
all together with other microscopies. In conclusion, SNOM, combined with rapid and
non-invasive methods of specimen preparation, appears to be a powerful tool that can
offer new possibilities in the field of neuroscience imaging at nano-scale level.