Second-harmonic generation and fluorescence lifetime imaging microscopy through a rodent mammary imaging window

Pamela A. Young; Muhammad Nazir; Michael J. Szulczewski; Patricia J. Keely; Kevin W. Eliceiri

doi:10.1117/12.906409

9 February 2012 Second-harmonic generation and fluorescence lifetime imaging microscopy through a rodent mammary imaging window

Pamela A. Young, Muhammad Nazir, Michael J. Szulczewski, Patricia J. Keely, Kevin W. Eliceiri

Author Affiliations +

Proceedings Volume 8226, Multiphoton Microscopy in the Biomedical Sciences XII; 822604 (2012) https://doi.org/10.1117/12.906409
Event: SPIE BiOS, 2012, San Francisco, California, United States

Abstract

Tumor-Associated Collagen Signatures (TACS) have been identified that manifest in specific ways during breast tumor progression and that correspond to patient outcome. There are also compelling metabolic changes associated with carcinoma invasion and progression. We have characterized the difference in the autofluorescent properties of metabolic co-factors, NADH and FAD, between normal and carcinoma breast cell lines. Also, we have shown in vitro that increased collagen density alters metabolic genes which are associated with glycolysis and leads to a more invasive phenotype. Establishing the relationship between collagen density, cellular metabolism, and metastasis in physiologically relevant cancer models is crucial for developing cancer therapies. To study cellular metabolism with respect to collagen density in vivo, we use multiphoton fluorescence excitation microscopy (MPM) in conjunction with a rodent mammary imaging window implanted in defined mouse cancer models. These models are ideal for the study of collagen changes in vivo, allowing determination of corresponding metabolic changes in breast cancer invasion and progression. To measure cellular metabolism, we collect fluorescence lifetime (FLIM) signatures of NADH and FAD, which are known to change based on the microenvironment of the cells. Additionally, MPM systems are capable of collecting second harmonic generation (SHG) signals which are a nonlinear optical property of collagen. Therefore, MPM, SHG, and FLIM are powerful tools with great potential for characterizing key features of breast carcinoma in vivo. Below we present the current efforts of our collaborative group to develop intravital approaches based on these imaging techniques to look at defined mouse mammary models.

Citation Download Citation

Pamela A. Young, Muhammad Nazir, Michael J. Szulczewski, Patricia J. Keely, and Kevin W. Eliceiri "Second-harmonic generation and fluorescence lifetime imaging microscopy through a rodent mammary imaging window", Proc. SPIE 8226, Multiphoton Microscopy in the Biomedical Sciences XII, 822604 (9 February 2012); https://doi.org/10.1117/12.906409

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