Analytical and Bioanalytical Chemistry

, Volume 403, Issue 8, pp 2197–2202

Pump-probe optical microscopy for imaging nonfluorescent chromophores


DOI: 10.1007/s00216-012-5890-1

Cite this article as:
Wei, L. & Min, W. Anal Bioanal Chem (2012) 403: 2197. doi:10.1007/s00216-012-5890-1


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.

Apparatus used for pump-probe microscopy


Pump-probe microscopyLabel-free imagingExcited state absorptionStimulated emissionGround state depletionNonfluorescent chromophore

Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  1. 1.Department of ChemistryColumbia UniversityNew YorkUSA