Device and methods for optical pathology

Abstract

Currently most cancers, including breast cancers, are removed without any intraoperative margin control. Post-operative methods inspect 1-2% of the surgical margin and are prone to sampling errors. The present invention relates to an optical imaging system that will enable evaluation of the surgical margin in vivo and in real-time. The invention provides for simultaneous fluorescence and fluorescence polarization imaging. The contrast of the acquired images will be enhanced using fluorescent agents approved for diagnostic use in patients. As the staining pattern of fluorescence images is similar to that of histology, and the values of fluorescence polarization are significantly higher in cancerous as compared to normal cells, the invention provides for further improvements in diagnostic methods. The systems and methods can be applied to the intra-operative delineation of cancerous tissue.

What is claimed is: 
     
         1 . A system for presenting images of a region of tissue, the system comprising:
 an illumination source configured to illuminate the region with first and second wavelengths of light;   one or more optical elements configured to image a confocal point of the second wavelength of light in the region of tissue and to scan the confocal point through a sub-region of the region;   a detector assembly configured to capture a wide-field fluorescence polarization image based on the first wavelength of light illuminating the region for grossly delineating a tumor margin in the region and to detect light emitted from the sub-region in response to the scanning of the confocal point of the second wavelength of light through the sub-region; and   a display configured to present the wide-field fluorescence polarization image and configured to present a confocal fluorescence polarization image of the sub-region, wherein the confocal fluorescence polarization image includes a resolved image of a cell in the sub-region, the confocal fluorescence polarization image being generated based on the light detected in response to the scanning of the confocal point through the sub-region,   wherein the displayed confocal fluorescence polarization image visually distinguishes, based on at least one quantitative value of a cellular characteristic of the cell, the cell as cancerous or non-cancerous, wherein the at least one quantitative value of the cellular characteristic is determined based on co-polarization and cross-polarization image values of pixels of the resolved image of the cell.   
     
     
         2 . A system for evaluating images of a cancer at a cellular level in a region of tissue, the system comprising:
 an illumination source configured to illuminate the region with light having a wavelength;   one or more optical elements configured to image a linearly-polarized confocal point of the light in the region of tissue and to scan the confocal point through a sub-region of the region;   a detector assembly configured to detect co-polarized and cross-polarized light emitted from the sub-region in response to the scanning of the confocal point of the second wavelength of light through the sub-region; and   a data processor connected to the detector assembly and configured to:
 generate a fluorescence polarization image of the sub-region based on the detected co-polarized and cross-polarized light, the fluorescence polarization image including a resolved image of a cell in the sub-region, wherein the generating includes determining, based on co-polarization and cross-polarization image values of pixels of the resolved image of the cell, an exogenous fluorescence polarization value for the cell, wherein the exogenous fluorescence polarization value indicates an intrinsic fluorescence polarization of the cell; and 
 a display configured to display the fluorescence polarization image, wherein the displayed image visually indicates, based on the exogenous fluorescence polarization value for the cell, whether the cell is cancerous or non-cancerous. 
   
     
     
         3 . A system as recited in  claim 2 , wherein the at least one exogenous fluorescence polarization value is significantly higher in a cancer cell as compared to a non-cancerous cell. 
     
     
         4 . A system as recited in  claim 2 , wherein the tissue includes a first contrast agent applied to the region as a cytoplasmic stain; and a second contrast agent applied to the region as a nuclear stain. 
     
     
         5 . A system as recited in  claim 4 , wherein, on the display in a single image: the cytoplasmic stain looks pink for cytoplasm, connective tissue and collagen; and the nuclear stain looks blue for cellular structure. 
     
     
         6 . A system as recited in  claim 2 , wherein the display is configured to selectively present a wide-field fluorescence polarization image and a high-resolution confocal reflectance image during surgery, in situ or during a biopsy.