Ovarian Imaging in Animal Models

We are mounting a three-dimensional OCT system inside the microscope to obtain co-registered OCT, TPEF, and SHG images. We will image mouse ovaries in our ex vivo study with the microscopic techniques, in addition to the OCT/LIF and OCM already planned. Since all modalities are in the same laboratory complex, and this supplement will provide an additional research specialist to perform imaging, the additional imaging will not require a lengthening of the overall study time. A comparison of the two OCT data sets will enable co-registration of LIF with the microscopy images, and we anticipate that the OCM images will show some of the same features seen in TPEF images, facilitating registration. Of particular interest, when comparing the image/spectra sets, will be discerning how the cellular and collagen structure visible in the microscopic images translates into the image texture we have noted in the lower resolution OCT images. This result will have implications for other work, such as our imaging of human ovary [Hariri et al., 2009], where for the time being, microscopic imaging techniques are less practical than OCT, and understanding how the OCT image texture relates to risk or disease status of the ovary could have important implications for high risk women considering oophorectomy . Similarly, we would like to determine how the TPEF images relate to point spectroscopy of LIF (which is likewise much easier to obtain in humans in vivo, and has been demonstrated by the investigators[Brewer et al., 2004]).

Finally, we will develop specialize animal holders than enable the mouse to be positioned and the ovary positioned for imaging, and have connections for gas anesthesia. Only the right ovary, which is easily accessible from a small incision in the flank, will be imaged. We will develop procedures to assure sterile imaging, and we will calculate image parameters (e.g. how many images in a stack) to keep microscopic imaging below 15 minutes. FLIM will be performed as discrete images at given depth(s) since this is the most time-consuming image type. We will incorporate the microscopic imaging into the existing in vivo imaging protocols. Images will be analyzed as in task 2, but additionally over time. The hope is that early image features will predict later neoplastic development.

References
         Brewer MA, Utzinger U, Barton JK, Hoying JB, Kirkpatrick ND, Brands WR, Davis JR, Hunt K, Stevens SJ, Gmitro AF, “Imaging of the ovary,” Technol Cancer Res Treat. 3(6):617-27, 2004.
          Hariri LP, Bonnema GT, Schmidt K, Winkler AM, Korde V, Hatch K, Brewer M, Barton JK “Laparoscopic optical coherence tomography imaging of human ovarian cancer,” accepted for publication in Gynecologic Oncology, 2009.
         Hoyer PB, Davis JR, Bedrnicek JB, Marion SL, Christian PJ, Barton JK, Brewer MA, "Ovarian neoplasm development by 7,12-dimethylbenz[a]anthracene (DMBA) in a chemically-induced rat model of ovarian failure,” Gynecological Oncology, 112:610615, 2009.
         Kirkpatrick ND, Brewer MA, Utzinger U, “Endogenous optical biomarkers of ovarian cancer evaluated with multiphoton microscopy,” Cancer Epidemiol Biomarkers Prev. 16(10):2048-57, 2007.
         Korde VR, Liebmann E, Barton JK, “Design of a handheld optical coherence microscopy endoscope,” Proceedings of the SPIE 7172:71720D, 2009.
         Skala, M., et al., In vivo Multiphoton Fluorescence Lifetime Imaging of Free and Protein-bound NADH in Normal and Pre-cancerous Epithelia. Optical Society of America: Biomedical Topical Meeting, 2006.