System and Method for Normalized Flourescence or Bioluminescence Imaging

Abstract

A system and method provide normalized fluorescence epi-illumination images and normalized fluorescence transillumination images. The normalization can be used to improve two-dimensional (planar) fluorescence epi-illumination images and two-dimensional (planar) fluorescence transillumination images. The system and method can also provide normalized bioluminescence epi-illumination images and normalized bioluminescence transillumination images. In some arrangements, the system and method can provide imagine of small animals, into-operative imaging, endoscopic imaging, and/or imaging of hollow organs.

1 . A method of imaging, comprising:
 generating incident light including excitation light with an excitation light source;   directing the incident light toward a tissue;   receiving the incident light with a light detector after the incident light has interacted with a tissue;   receiving emitted light with the light detector, wherein the emitted light is emitted from the tissue;   generating an intrinsic image of the tissue in response to the incident light;   generating an un-normalized emitted light image of the tissue, and   combining the un-normalized emitted light image and the intrinsic image to generate a normalized emitted light image of the tissue.   
     
     
         2 . The method of  claim 1 , further comprising: inserting a probe into a patient during an operative procedure, wherein the generating the incident light includes transmitting the incident light from the probe, wherein the receiving the incident light includes receiving the incident light with the probe, and wherein the receiving the emitted light includes receiving the emitted light with the probe. 
     
     
         3 . The method of  claim 2 , wherein the operative procedure comprises imaging an internal body cavity of a patient. 
     
     
         4 . The method of  claim 1 , wherein the method comprises a selected one of a laparoscopic imaging method, an endoscopic imaging method, a colonoscopic imaging method, a colposcopic imaging method, an esophageal imaging, method, a pulmonary imaging method, an oral imaging method, and a dental imaging method. 
     
     
         5 . The method of  claim 1 , wherein:
 the excitation light source comprises an epi-illumination light source disposed on generally the same side of the tissue as the light detector;   the emitted light comprises fluorescent light associated with the tissue;   the intrinsic image comprises an intrinsic epi-illumination image;   the un-normalized emitted light image comprises an un-normalized fluorescence epi-illumination image; and   the normalized emitted light image comprises a normalized fluorescence epi-illumination image.   
     
     
         6 . The method of  claim 5 , wherein fluorescent light is generated by at least one of a fluorescent marker and an endogenous tissue fluorescent molecule in response to the excitation light, wherein the fluorescent marker is administered to the tissue. 
     
     
         7 . The method of  claim 6 , wherein the incident light is substantially at one wavelength. 
     
     
         8 . The method of  claim 1 , wherein:
 the excitation light source comprises a transillumination light source disposed on generally the opposite side of the tissue as the light detector;   the emitted light comprises fluorescent light associated with the tissue;   the intrinsic image comprises an intrinsic transillumination image;   the un-normalized emitted light image comprises an un-normalized fluorescence transillumination image; and   the normalized emitted light image comprises a normalized fluorescence transillumination image.   
     
     
         9 . The method of  claim 8 , wherein the fluorescent light is generated by at least one of a fluorescent marker and an endogenous tissue fluorescent molecule in response to the excitation light, wherein the fluorescent marker is administered to the tissue. 
     
     
         10 . The method of  claim 9 , wherein the incident light is substantially at one wavelength. 
     
     
         11 . The method of  claim 8 , wherein the transillumination light source comprises a plurality of individual transillumination light sources, wherein the method further comprises:
 combining a plurality of intrinsic images to generate the intrinsic image, wherein each one of the plurality of intrinsic images is associated with a respective one of the plurality of transillumination light sources, and   combining a plurality of un-normalized emitted light images to generate the un-normalized emitted light image, wherein each one of the plurality of un-normalized emitted light images is associated with a respective one of the plurality of transillumination light sources.   
     
     
         12 . The method of  claim 1 , wherein:
 the excitation light source comprises an epi-illumination light source disposed on generally the same side of the tissue as the light detector;   the emitted light comprises bioluminescence light associated with the tissue;   the intrinsic image comprises an intrinsic epi-illumination image;   the un-normalized emitted light image comprises and un-normalized bioluminescence image; and   the normalized emitted light image comprises a normalized bioluminescence epi-illumination image.   
     
     
         13 . The method of  claim 1 , wherein:
 the excitation light source comprises a transillumination light source disposed on generally the opposite side of the tissue as the light detector;   the emitted light comprises bioluminescence light associated with the tissue;   the intrinsic image comprises an intrinsic transillumination image;   the un-normalized emitted light image comprises an un-normalized bioluminescence image; and   the normalized emitted light image comprises a normalized bioluminescence epi-illumination image.   
     
     
         14 . The method of  claim 13 , wherein the transillumination light source comprises a plurality of individual transillumination light sources, wherein the method further comprises:
 combining a plurality of intrinsic images to generate the intrinsic image, wherein each one of the plurality of intrinsic images is associated with a respective one of the plurality of transillumination light sources.   
     
     
         15 . The method of  claim 1 , wherein the combining comprises dividing the un-normalized emitted light image by the intrinsic image. 
     
     
         16 . The method of  claim 15 , wherein the dividing comprises dividing pixel magnitudes of pixels of the un-normalized emitted light image by pixel magnitudes of co-registered pixels of the intrinsic image. 
     
     
         17 . The method of  claim 1 , wherein the incident light has a wavelength in the range of four hundred to one thousand nanometers and the emitted light also has a wavelength in the range of four hundred to one thousand nanometers, wherein the wavelength of the emitted light is shorter than the wavelength of the excitation light. 
     
     
         18 . The method of  claim 1 , wherein the light receiver comprises a charge coupled device camera. 
     
     
         19 . The method of  claim 1 , further including:
 generating a first background image of the tissue at generally the same wavelength as the intrinsic image;   generating a second background image of the tissue at generally the same wavelength as the un-normalized emitted light image;   combining the intrinsic image and the first background image to provide a noise-reduced intrinsic image;   combining the un-normalized emitted light image and the second background image to provide a noise-reduced un-normalized emitted light image; and   combining the noise-reduced un-normalized emitted light image and the noise-reduced intrinsic image to provide a noise-reduced normalized emitted light image.   
     
     
         20 . The method of  claim 1 , wherein the incident light includes one or more wavelengths, and wherein the intrinsic image is generated by combining intrinsic images associated with the one or more wavelengths. 
     
     
         21 . A system for imaging a tissue, the system comprising:
 an excitation light source adapted to generate incident light including excitation light;   a light receiver adapted to receive the incident light after the incident light has interacted with the tissue, further adapted to receive emitted light, wherein the emitted light is emitted from the tissue, further adapted to generate an intrinsic image of the tissue in response to the incident light, and further adapted to generate an un-normalized emitted light image of the tissue, and   a normalization processor adapted to combine the un-normalized emitted light image and the intrinsic image to generate a normalized emitted light image associated with the tissue.   
     
     
         22 . The system of  claim 21 , further comprising:
 a probe; and   at least one optical fiber coupled between the probe and the excitation light source and between the probe and the light receiver, wherein the at least one optical fiber is adapted to carry the incident light to the probe, to receive the incident light with the probe after the incident light has interacted with the tissue, and to receive the emitted light with the probe.   
     
     
         23 . The system of  claim 22 , wherein the probe is adapted to image an internal body cavity of a patient. 
     
     
         24 . The system of  claim 22 , wherein probe comprises a selected one of a laparoscopic probe, an endoscopic probe, a colonoscopic probe, a colposcopic probe, an esophageal probe, a pulmonary probe, an oral probe, and a dental probe. 
     
     
         25 . The system of  claim 21 , wherein:
 the excitation light source comprises an epi-illumination light source disposed on generally the same side of the tissue as the light detector;   the emitted light comprises fluorescent light associated with the tissue;   the intrinsic image comprises an intrinsic epi-illumination image;   the un-normalized emitted light image comprises an un-normalized fluorescence epi-illumination image; and   wherein the normalization processor comprises a fluorescence epi-illumination normalization processor adapted to generate the normalized emitted light image as a normalized fluorescence epi-illumination image.   
     
     
         26 . The system of  claim 25 , wherein fluorescent light is generated by at least one of a fluorescent marker and an endogenous tissue fluorescent molecule in response to the excitation light, wherein the fluorescent marker is administered to the tissue. 
     
     
         27 . The system of  claim 26 , wherein the incident light is substantially at one wavelength. 
     
     
         28 . The system of  claim 21 , wherein:
 the excitation light source comprises a transillumination light source disposed on generally the opposite side of the tissue as the light detector;   the emitted light comprises fluorescent light associated with the tissue;   the intrinsic image comprises an intrinsic transillumination image;   the un-normalized emitted light image comprises an un-normalized fluorescence transillumination image; and   wherein the normalization processor comprises a fluorescence transillumination normalization processor adapted to generate the normalized emitted light image as a normalized fluorescence transillumination image.   
     
     
         29 . The system of  claim 28 , wherein fluorescent light is generated by at least one of a fluorescent marker and an endogenous tissue fluorescent molecule in response to the excitation light, wherein the fluorescent marker is administered to the tissue. 
     
     
         30 . The system of  claim 29 , wherein the incident light is substantially at one wavelength. 
     
     
         31 . The system of  claim 28 , wherein the transillumination light source comprises a plurality of individual transillumination light sources, wherein the normalization processor comprises:
 an intrinsic image combining processor adapted to combine a plurality of intrinsic images to generate the intrinsic image, wherein each one of the plurality of intrinsic images is associated with a respective one of the plurality of transillumination light sources; and   an emitted image combining processor adapted to combine a plurality of un-normalized emitted light images to generate the un-normalized emitted light image, wherein each one of the plurality of un-normalized emitted light images is associated with a respective one of the plurality of transillumination light sources.   
     
     
         32 . The system of  claim 21 , wherein:
 the excitation light source comprises an epi-illumination light source disposed on generally the same side of the tissue as the light detector;   the emitted light comprises bioluminescence light associated with the tissue;   the intrinsic image comprises an intrinsic epi-illumination image;   the un-normalized emitted light image comprises and un-normalized bioluminescence image; and   wherein the normalization processor comprises a bioluminescence epi-illumination normalization processor adapted to generate the normalized emitted light image as a normalized bioluminescence epi-illumination image.   
     
     
         33 . The system of  claim 21 , wherein:
 the excitation light source comprises a transillumination light source disposed on generally the opposite side of the tissue as the light detector;   the emitted light comprises bioluminescence light associated with the tissue;   the intrinsic image comprises an intrinsic transillumination image;   the un-normalized emitted light image comprises an un-normalized bioluminescence image; and   wherein the normalization processor comprises a bioluminescence transillumination normalization processor adapted to generate the normalized emitted light image as a normalized bioluminescence transillumination image.   
     
     
         34 . The system of  claim 33 , wherein the transillumination light source comprises a plurality of individual transillumination light sources, wherein the normalization processor comprises an intrinsic excitation image combining processor adapted to combine a plurality of intrinsic images to generate the intrinsic image, wherein each one of the plurality of intrinsic images is associated with a respective one of the plurality of transillumination light sources. 
     
     
         35 . The system of  claim 21 , wherein the normalization processor is adapted to combine the un-normalized emitted light image and the intrinsic image by dividing the un-normalized emitted light image by the intrinsic image. 
     
     
         36 . The method of  claim 35 , wherein the normalization processor is adapted to combine the un-normalized emitted light image and the intrinsic image by dividing pixel magnitudes of pixels of the un-normalized emitted light image by pixel magnitudes of co-registered pixels of the intrinsic image. 
     
     
         37 . The system of  claim 21 , wherein the incident light has a wavelength in the range of four hundred to one thousand nanometers and the emitted light also has a wavelength in the range off four hundred to one thousand nanometers, wherein the wavelength of the emitted light is shorter than the wavelength of the excitation light. 
     
     
         38 . The system of  claim 21 , wherein the light receiver comprises a charge coupled device camera. 
     
     
         39 . The system of  claim 21 , further including:
 a first optical filter coupled to the light detector and adapted to pass the excitation light, wherein the excitation light has a first wavelength; and   a second optical filter coupled to the light detector and adapted to pass the emitted light, wherein the emitted light has a second wavelength different from the first wavelength.   
     
     
         40 . The system of  claim 21 , further including an optical filter coupled to the light detector and adapted to pass the excitation light and the emitted light. 
     
     
         41 . The system of  claim 21 , wherein the light detector is further adapted to generate a first background image of the tissue at generally the same wavelength as the intrinsic image and adapted to generate a second background image of the tissue at generally the same wavelength as the un-normalized emitted light image, and wherein the normalization processor comprises:
 an intrinsic image noise reduction processor adapted to combine the intrinsic image and the first background image to provide a noise-reduced intrinsic image; and   an emitted image noise reduction processor adapted to combine the un-normalized emitted light image and the second background image to provide a noise-reduced un-normalized emitted light image, wherein normalization processor is further adapted to combine the noise-reduced un-normalized emitted light image and the noise-reduced intrinsic image to provide a noise-reduced normalized emitted light image.   
     
     
         42 . The system of  claim 21 , wherein the incident light includes one or more wavelengths, and wherein the intrinsic image is generated by combining intrinsic images associated with the one or more wavelengths.