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Published June 2012 | public
Journal Article

Pump-probe optical microscopy for imaging nonfluorescent chromophores

Abstract

Many chromophores absorb light intensely but have undetectable fluorescence. Hence microscopy techniques other than fluorescence are highly desirable for imaging these chromophores inside live cells, tissues, and organisms. The recently developed pump-probe optical microscopy techniques provide fluorescence-free contrast mechanisms by employing several fundamental light–molecule interactions including excited state absorption, stimulated emission, ground state depletion, and the photothermal effect. By using the pump pulse to excite molecules and the subsequent probe pulse to interrogate the created transient states on a laser scanning microscope, pump-probe microscopy offers imaging capability with high sensitivity and specificity toward nonfluorescent chromophores. Single-molecule sensitivity has even been demonstrated. Here we review and summarize the underlying principles of this emerging class of molecular imaging techniques.

Additional Information

© Springer-Verlag 2012. Received: 5 December 2011 /Revised: 16 February 2012 / Accepted: 20 February 2012 / Published online: 13 March 2012. We acknowledge discussions with X. S. Xie, C. W. Freudiger, S. Lu, S. Chong, B. G. Saar, G. R. Holtom, M. Roeffaers, D. Fu, X. Zhang, and R. Roy. Published in the special issue Young Investigators in Analytical and Bioanalytical Science with Guest Editors S. Daunert, J. Bettmer, T. Hasegawa, Q. Wang and Y. Wei.

Additional details

Created:
August 19, 2023
Modified:
October 19, 2023