Microscopy and Microanalysis

Multiphoton Microscopy–Special Section

Nonlinear Optical Imaging of Cellular Processes in Breast Cancer

Paolo P. Provenzanoa1a2a3, Kevin W. Eliceiria2 c1, Long Yana2, Aude Ada-Nguemaa1, Matthew W. Conklina1a2a3, David R. Inmana1a3 and Patricia J. Keelya1a2a3

a1 Department of Pharmacology, University of Wisconsin, Madison, WI 53706

a2 Laboratory for Optical and Computational Instrumentation, University of Wisconsin, Madison, WI 53706

a3 University of Wisconsin Comprehensive Cancer Center, University of Wisconsin, Madison, WI 53792


Nonlinear optical imaging techniques such as multiphoton and second harmonic generation (SHG) microscopy used in conjunction with novel signal analysis techniques such as spectroscopic and fluorescence excited state lifetime detection have begun to be used widely for biological studies. This is largely due to their promise to noninvasively monitor the intracellular processes of a cell together with the cell's interaction with its microenvironment. Compared to other optical methods these modalities provide superior depth penetration and viability and have the additional advantage in that they are compatible technologies that can be applied simultaneously. Therefore, application of these nonlinear optical approaches to the study of breast cancer holds particular promise as these techniques can be used to image exogeneous fluorophores such as green fluorescent protein as well as intrinsic signals such as SHG from collagen and endogenous fluorescence from nicotinamide adenine dinucleotide or flavin adenine dinucleotide. In this article the application of multiphoton excitation, SHG, and fluorescence lifetime imaging microscopy to relevant issues regarding the tumor-stromal interaction, cellular metabolism, and cell signaling in breast cancer is described. Furthermore, the ability to record and monitor the intrinsic fluorescence and SHG signals provides a unique tool for researchers to understand key events in cancer progression in its natural context.

(Received July 11 2008)

(Accepted July 21 2008)


c1 Corresponding author. E-mail: eliceiri@wisc.edu