Wisconsin Alumni Research Foundation

John Graham White

Kevin Eliceiri

Paolo Pepe Provenzano

Patricia Jo Keely

David L Wokosin

The present invention broadly provides methods and systems for detecting, identifying, and characterizing conditions, including diseases and other disorders in human or other animal subjects, by analyzing fluorescence from endogenous flavin adenine dinucleotide (FAD) fluorophors present in biological materials and samples. In particular embodiments, the invention relates to conditions of the human breast including cancers such as carcinoma. Methods and systems are provided for detecting, locating, and characterizing tumors and precancerous tissue via nonlinear optical imaging techniques capable of accurately characterizing fluorescence intensities and fluorescent lifetime parameters from endogenous FAD fluorophors present in a test tissue sample.

Use of endogenous fluorescence to identify invading metastatic breast tumor cells

The present invention provides methods and systems for evaluating biological materials for the diagnosis of disease, such as gland abnormalities, and the cancerous and precancerous conditions. Nonlinear optical microscopy techniques, such as MP microscopy and harmonic generation microscopy, are used to generate high resolution, three dimensional images of a test tissue, such as a biopsy tissue sample and tissue in whole organisms, that are analyzed, optionally in combination, to detect, identify and characterize tumor-associated collagen signatures. The presence, abundance and extent of histological features and structural motifs comprising tumor-associated collagen signatures may be directly and accurately correlated with the onset and progression of cancer, such as breast cancer. The present methods are capable of providing an accurate and selective diagnosis of cancer, and provide diagnostic information complementary to conventional diagnostic methods.

Stromal collagen in the diagnosis and characterization of breast cancer

A photon sorting spectroscopic microscope system focuses light from an excitation source onto a specimen through a microscope objective lens to excite fluorescence in the specimen. The fluorescent light is passed back through the objective on a beam path and is directed by a dichroic mirror on a second beam path to a spectral dispersive element which spreads the light according to its spectral content. A multi-channel detector with multiple detector elements receives the light spread by the spectral dispersive element and provides output signals indicative of photon events. The photon events in each channel may be counted to provide the spectral content of the fluorescent signal as the excitation beam is scanned in a raster fashion over the sample. Two dimensional fluorescence lifetime analysis may further be provided by utilizing multiple counters on each channel which are sequentially enabled after a pulse of light from the source, providing photon counts in each spectral channel as a function of time delay from the excitation light pulse.

Photon-sorting spectroscopic microscope system

Multi-photon excitation fluorescence microscopy is carried out by focusing an excitation beam by an objective lens onto a specimen and collecting the multi-photon fluorescence light emitted from the specimen to the objective lens and directing the fluorescent light on an optical path to a detector. Fluorescent light emissions from the specimen collected by a condenser lens on the opposite side of the specimen from the objective lens are directed to a dichoic mirror, which reflects the light photons back into the condenser lens and thence into and through the objective lens where they are directed on an optical path to the detector. Significantly increased fluorescent photon collection efficiency is obtained as well as improved image intensity of the detected fluorescent light.

Growing community of inventors

Madison, WI, United States of America

John Graham White