162
C. Li and J.-X. Cheng
6.4 Conclusion and Outlook
The functions of biomolecules, biosystems, and nanomaterials are inextricably linked
with the structures and chemical compositions. Absorption-based LFSRM, with
superior detection limit and spatial resolution, is growing as a significant tool to
enhance the understanding of biology and materials sciences. In this review, we have
introduced both linear and nonlinear absorption-based LFSRM techniques that provide excellent sensitivity, fast imaging speed, as well as informative spectroscopic
information. We have highlighted how the recent advancements in mid-IR excited
PTM have successfully defeated the diffraction limit of mid-IR beam. As a relatively
new field, it is to be expected that absorption-based LFSRM approaches will be further improved by ongoing developments in laser engineering, optical and electronic
technology.
References
1. J.B. Pawley, in Handbook of Biological Confocal Microscopy, ed. by J.B. Pawley (Springer
US, Boston, MA, 2006), pp. 20–42
2. E. Abbe, Beiträge zur Theorie des Mikroskops und der mikroskopischen Wahrnehmung.
Archiv für mikroskopische Anatomie 9, 413–418 (1873)
3. S.W. Hell, Microscopy and its focal switch. Nat. Methods 6, 24–32 (2009)
4. R.M. Dickson, A.B. Cubitt, R.Y. Tsien, W.J.N. Moerner, On/off blinking and switching
behaviour of single molecules of green fluorescent protein. Nature 388, 355–358 (1997)
5. E. Betzig, G.H. Patterson, R. Sougrat, O.W. Lindwasser, S. Olenych, J.S. Bonifacino, M.W.
Davidson, J. Lippincott-Schwartz, H.F.J.S. Hess, Imaging intracellular fluorescent proteins at
nanometer resolution. Science 313, 1642–1645 (2006)
6. M.J. Rust, M. Bates, X. Zhuang, Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM). Nat. Methods 3, 793–795 (2006)
7. B. Huang, W. Wang, M. Bates, X.J.S. Zhuang, Three-dimensional super-resolution imaging
by stochastic optical reconstruction microscopy. Science 319, 810–813 (2008)
8. S.W. Hell, J.J.O.L. Wichmann, Breaking the diffraction resolution limit by stimulated emission: stimulated-emission-depletion fluorescence microscopy. Opt. Lett. 19, 780–782 (1994)
9. K.I. Willig, B. Harke, R. Medda, S.W. Hell, STED microscopy with continuous wave beams.
Nat. Methods 4, 915–918 (2007)
10. K.I. Willig, S.O. Rizzoli, V. Westphal, R. Jahn, S.W. Hell, STED microscopy reveals that
synaptotagmin remains clustered after synaptic vesicle exocytosis. Nature 440, 935–939
(2006)
11. S.W. Hell, M.J.A.P.B. Kroug, Ground-state-depletion fluorscence microscopy: a concept for
breaking the diffraction resolution limit. Appl. Phys. B 60, 495–497 (1995)
12. M.G. Gustafsson, Surpassing the lateral resolution limit by a factor of two using structured
illumination microscopy. J. Microsc. 198, 82–87 (2000)
13. M.G. Gustafsson, Nonlinear structured-illumination microscopy: wide-field fluorescence
imaging with theoretically unlimited resolution. Proc. Natl. Acad. Sci. U.S.A. 102,
13081–13086 (2005)
14. J.-X. Cheng, X.S. Xie, Coherent anti-Stokes Raman scattering microscopy: instrumentation,
theory, and applications. J. Phys. Chem. B 108, 827–840 (2004)
15. C.W. Freudiger, W. Min, B.G. Saar, S. Lu, G.R. Holtom, C. He, J.C. Tsai, J.X. Kang, X.S.J.S.
Xie, Label-free biomedical imaging with high sensitivity by stimulated Raman scattering
microscopy. Science 322, 1857–1861 (2008)
C. Li and J.-X. Cheng
6.4 Conclusion and Outlook
The functions of biomolecules, biosystems, and nanomaterials are inextricably linked
with the structures and chemical compositions. Absorption-based LFSRM, with
superior detection limit and spatial resolution, is growing as a significant tool to
enhance the understanding of biology and materials sciences. In this review, we have
introduced both linear and nonlinear absorption-based LFSRM techniques that provide excellent sensitivity, fast imaging speed, as well as informative spectroscopic
information. We have highlighted how the recent advancements in mid-IR excited
PTM have successfully defeated the diffraction limit of mid-IR beam. As a relatively
new field, it is to be expected that absorption-based LFSRM approaches will be further improved by ongoing developments in laser engineering, optical and electronic
technology.
References
1. J.B. Pawley, in Handbook of Biological Confocal Microscopy, ed. by J.B. Pawley (Springer
US, Boston, MA, 2006), pp. 20–42
2. E. Abbe, Beiträge zur Theorie des Mikroskops und der mikroskopischen Wahrnehmung.
Archiv für mikroskopische Anatomie 9, 413–418 (1873)
3. S.W. Hell, Microscopy and its focal switch. Nat. Methods 6, 24–32 (2009)
4. R.M. Dickson, A.B. Cubitt, R.Y. Tsien, W.J.N. Moerner, On/off blinking and switching
behaviour of single molecules of green fluorescent protein. Nature 388, 355–358 (1997)
5. E. Betzig, G.H. Patterson, R. Sougrat, O.W. Lindwasser, S. Olenych, J.S. Bonifacino, M.W.
Davidson, J. Lippincott-Schwartz, H.F.J.S. Hess, Imaging intracellular fluorescent proteins at
nanometer resolution. Science 313, 1642–1645 (2006)
6. M.J. Rust, M. Bates, X. Zhuang, Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM). Nat. Methods 3, 793–795 (2006)
7. B. Huang, W. Wang, M. Bates, X.J.S. Zhuang, Three-dimensional super-resolution imaging
by stochastic optical reconstruction microscopy. Science 319, 810–813 (2008)
8. S.W. Hell, J.J.O.L. Wichmann, Breaking the diffraction resolution limit by stimulated emission: stimulated-emission-depletion fluorescence microscopy. Opt. Lett. 19, 780–782 (1994)
9. K.I. Willig, B. Harke, R. Medda, S.W. Hell, STED microscopy with continuous wave beams.
Nat. Methods 4, 915–918 (2007)
10. K.I. Willig, S.O. Rizzoli, V. Westphal, R. Jahn, S.W. Hell, STED microscopy reveals that
synaptotagmin remains clustered after synaptic vesicle exocytosis. Nature 440, 935–939
(2006)
11. S.W. Hell, M.J.A.P.B. Kroug, Ground-state-depletion fluorscence microscopy: a concept for
breaking the diffraction resolution limit. Appl. Phys. B 60, 495–497 (1995)
12. M.G. Gustafsson, Surpassing the lateral resolution limit by a factor of two using structured
illumination microscopy. J. Microsc. 198, 82–87 (2000)
13. M.G. Gustafsson, Nonlinear structured-illumination microscopy: wide-field fluorescence
imaging with theoretically unlimited resolution. Proc. Natl. Acad. Sci. U.S.A. 102,
13081–13086 (2005)
14. J.-X. Cheng, X.S. Xie, Coherent anti-Stokes Raman scattering microscopy: instrumentation,
theory, and applications. J. Phys. Chem. B 108, 827–840 (2004)
15. C.W. Freudiger, W. Min, B.G. Saar, S. Lu, G.R. Holtom, C. He, J.C. Tsai, J.X. Kang, X.S.J.S.
Xie, Label-free biomedical imaging with high sensitivity by stimulated Raman scattering
microscopy. Science 322, 1857–1861 (2008)