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observation of dynamic biomolecular processes is straightforward and
can give deep insights into their functional mechanisms. Therefore, this
new microscopy will markedly change our style of considering biological
questions. Nevertheless, there are presently only few setups of high-speed
bio-AFM that can capture dynamic biomolecular processes at 10-30 frames/
s, and consequently, the user population is limited. Besides, to our knowledge,
only two manufacturers are producing small cantilevers for high-speed
bio-AFM. We hope that this current situation will be quickly improved by
manufacturers.
In the near future, high-speed AFM will be actively used to observe a
wide range of dynamic processes that occur on isolated proteins, protein
assemblies and protein-DNA complexes. More complex systems including
live cells and organisms will become targets of high-speed AFM after some
technical advances described earlier are successfully overcome. The in vivo
and in vitro visualization of various processes at the molecular level will
become possible including the responses of membrane receptors to stimuli,
nuclear envelope formation and disassembly, chromosome replication and
segregation processes, phagocytosis, protein synthesis in the endoplasmic
reticulum and the targeting processes of synthesized proteins through the
Golgi apparatus. Thus, high-speed AFM-based visualization techniques
have great potential to bring about breakthroughs not only in biochemistry
and biophysics but also in cell biology, physiology and pharmaceutical and
medical sciences. To open up such unprecedented ields, steady efforts have
to be carried out towards expanding the capability of high-speed AFM and
related techniques.
Acknowledgements
We thank D. Yamamoto, N. Kodera, M. Shibata, H. Yamashita and all previous
students for their dedicated studies for developing high-speed AFM. This work
was partially supported by the Japan Science, Technology Agency (JST; the
CREST program and a Grant-in-Aid for Development of Systems, Technology
for Advanced Measurement and Analysis and Strategic International
Cooperative Program), the Japan Society for the Promotion of Science (JSPS;
a Grant-in-Aid for Basic Research (S), Grant-in-Aid for Science Research on
Priority Areas; innovative nanoscience of supramolecular motor proteins
working in biomembranes), industrial technology research grant program in
'04 from New Energy and Industrial Technology Development Organization
(NEDO).
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