In order to determine
the biological origin of the optical signatures
of intracellular and extracellular constituents
we propose to further develop and study an
in-vitro tissue slice model.
Biopsies will be obtained from women undergoing
oophorectomy.. Embedded tissue samples will
be sectioned with a precision cut tissue slicer,
incubated, and microscopically examined. We
will determine the duration of the tissue
slice viability. Development of the ovarian
tissue slice model will be in collaboration
with investigators from the University of
Arizona, College of Pharmacy, and will have
application in many future studies.
We will explore the optical signature of tissue
constituents, quantify them and mapping their
spatial distribution within the tissue slice
with microscopy techniques. Images will be
obtained with white light and two-photon excitation
microscopy. White light reflectance images
will provide scattering information while
fluorescence images will determine spatial
distribution of fluorophores. Optical properties
from structures identified in these images
will be correlated with histopathologic analysis.
The focus will be to identify structures contributing
to the contrast between normal and neoplastic
transformed ovarian tissue. Imaging will be
performed at the University of Arizona in
the user facilities of the Arizona Research
Our efforts to achieve aim 1 and 2 require
data from biopsy samples from pre and postmenopausal
women because hormonal status will affect
tissue composition. Our samples will be studied
ex-vivo because our in-vitro tissue model
will keep tissue alive during microscopy studies.
Figure: Single photon and two
photon images of the ovarian surface epithelium
as well as the stroma. Left: 2P excitation
at 720 nm and fluorescence emission recorded
after IR filtering. Strongest signal is originating
from the epithelial tissue layer. Structural
contrast can also be observed in the stroma.
Right: The same site excited at 458 nm in
single photon mode shows stronger contribution
from the epithelium and less structural contrast.
Emission at 458nm excitation is consistent
with FAD and emission from 2P excitation at
720nm with NAD(P)H and structural proteins.
A bar is 200 mm.