Dr. Pamela A. Young Profile

Dr. Pamela A. Young

Postdoctoral Fellow at Lab for Optical and Computational Instrumentation

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Proceedings Article | 9 February 2012 Paper

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

Pamela Young, Muhammad Nazir, Michael Szulczewski, Patricia Keely, Kevin Eliceiri

Proceedings Volume 8226, 822604 (2012) https://doi.org/10.1117/12.906409

KEYWORDS: Fluorescence lifetime imaging, Collagen, Tumors, Luminescence, Second-harmonic generation, Microscopy, Cancer, Breast, Mode conditioning cables, In vivo imaging

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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.

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