Photon structure and chemometrics pathologic system

A method for quantifying a molecular content of a biological tissue by utilizing the auto-fluorescence thereof, comprising steps of

• (1) imaging native fluorescence of a tissue section with a fluorescence imaging microscope at multiple excitation wavelengths to obtain a fluorescence microscope image, and
• (2) computing and factorizing the fluorescence microscope image by the following equation to obtain a tissue native fluorescent component data map:
• (
• I
• 1
• I
• 2
• M
• I
• m
• )
• =
• (
• w
• 11
• w
• 12
• Λ
• w
• 1
• ⁢
• n
• w
• 21
• w
• 22
• Λ
• w
• 2
• ⁢
• n
• M
• M
• O
• M
• w
• m
• ⁢
• ⁢
• 1
• w
• m
• ⁢
• ⁢
• 2
• Λ
• w
• mn
• )
• ⁢
• (
• C
• 1
• C
• 2
• M
• C
• m
• )
• ,
• where I is the fluorescence intensity, w is a weight factor, C is the content of a fluorescent substance, weight wij can be regarded as the spectral signature of the jth component, the weights are normalized by Σiwij=1 to ensure that the total fluorescence intensity map produced by the jth component is Cj, the weights can be estimated from measurement of “pure” fluorescent components under the identical experimental condition noting that the weights may differ slightly between pure fluorescence and fluorophores in tissue owing to their different chemical environments, a non-negative matrix factorization (NMF) of the measured RGB channels may be utilized to extract both the weights and the fluorescence map directly from experimental data, the fluorescence map Cj is identified to be one of the fluorescence species with the spectral signature matching and a priori information on their spatial distribution characteristics, and then the absolute concentration and distribution of fluorescent molecules is given by Cj/Kj, where Kj a calibrated constant representing the total fluorescence, i.e. the fluorescence intensity produced by one mole of the pure jth component under the identical experimental condition, wherein the tissue native fluorescent component data map is the absolute concentration.

A method for performing cancer diagnosis and prognosis on unlabeled pathology tissue sections by using a photonic structural and chemometric pathology system, comprising

• obtaining a tissue sample from a human patient,
• preparing an unlabeled pathology section from the tissue sample,
• obtaining a phase differential microscope (Q-DIC) image of the pathology section,
• computing one or more two-dimensional Q-DIC quantitative maps from the Q-DIC image,
• obtaining a fluorescence microscope image of the pathology section at multiple excitation wavelengths,
• computing a fluorescent component data map from the fluorescence microscope image,
• generating or obtaining a risk scoring methodology comprising correlations based on statistical analysis of a database of two-dimensional Q-DIC quantitative maps and fluorescent component data maps for subjects with known pathology, and
• applying the risk scoring methodology to the combination of the two-dimensional Q-DIC quantitative maps and the fluorescent component data maps to obtain a cancer diagnosis and prognosis for the patient based on the patient's tissue sample and the database of two-dimensional Q-DIC quantitative maps and fluorescent component data maps for subjects with known pathology.