High resolution microendoscope employing differential structured illumination and method of using same

Abstract

A microendoscope system configured to create a sectioned image can include a spinning disc with a reflective grating. At least a portion of light returning from a fiber bundle and that passes through the grating can be imaged onto a CCD. At least a portion of light returning from the fiber bundle and which strikes the reflective grating can emanate from the background of the sample being imaged. This is reflected onto a second CCD, thereby resulting in a recorded background image. Subtracting the two images can result in a sectioned image.

What is claimed is: 
     
         1 . A high-resolution microendoscope system comprising:
 a light source;   a fiber optic bundle configured to transmit light from the light source to a sample;   a disc configured to receive light returned from the sample, the disc having spaced-apart segments, the spaced-apart segments being at least one of openings and transparent portions;   a first camera configured to capture a first image based at least in part on light passing through the disc; and   a second camera configured to capture a second image based at least in part on light reflected from the disc.   
     
     
         2 . The microendoscope system of  claim 1 , wherein the second image is subtracted from the first image to create an optically sectioned image. 
     
     
         3 . The microendoscope system of  claim 1 , wherein subtracting the second image from the first image allows nuclear morphology to be more easily discernable to a user than when viewing the first image alone. 
     
     
         4 . The microendoscope system of  claim 1 , wherein the first and second images are recorded simultaneously, and wherein the first image is a widefield image. 
     
     
         5 . The microendoscope system of  claim 1 , further comprising:
 a motor operatively connected to the disc and configured to rotate the disc; and   at least one processor operatively connected to both the first and second cameras and the motor.   
     
     
         6 . The microendoscope system of  claim 1 , wherein the microendoscope is configured to image fluorescent targets in vivo. 
     
     
         7 . The microendoscope system of  claim 1 , wherein the microendoscope is configured to perform optical sectioning in real-time at video rates. 
     
     
         8 . The microendoscope system of  claim 1 , wherein each camera is a charge-coupled device (CCD) camera, wherein the first camera is positioned on a first side of the disc and the second camera is positioned on an opposing second side of the disc. 
     
     
         9 . The microendoscope system of  claim 1 , further comprising:
 a first lens;   a second lens; and   an emission filter,   wherein each of the first lens, the second lens and the emission filter are positioned between the disc and the second camera.   
     
     
         10 . A method of using a high-resolution microendoscope system, the method comprising:
 capturing a first image with a first camera:   capturing a second image with a second camera spaced-apart from the first camera; and   creating an optically sectioned image by subtracting the second image from the first image.   
     
     
         11 . The method of  claim 10 , wherein each of the first and second cameras are a charge-coupled device (CCD) camera. 
     
     
         12 . The method of  claim 10 , further comprising:
 illuminating a sample and receiving light reflected from the sample prior to capturing the first and second images.   
     
     
         13 . The method of  claim 12 , wherein the sample is illuminated by a light emitting diode lamp and a fiber optic bundle. 
     
     
         14 . The method of  claim 12 , further comprising:
 rotating a disc to reflect light to the second camera and permit light to pass to the first camera prior to capturing the first and second images.   
     
     
         15 . A system comprising:
 one or more processors; and   one or more memories operatively coupled to at least one of the one or more processors and having instructions stored thereon that, when executed by at least one of the one or more processors, cause at least one of the one or more processors to:
 illuminate a sample; 
 rotate a disc to reflect certain light received from the sample and permit certain light received from the sample to pass through one or more portions of the disc; 
 capture a first image with a first camera based at least in part on light passing through one or more portions of the disc; 
 capture a second image with a second camera based at least in part on light reflected from one or more portions of the disc, the second camera being spaced-apart from the first camera; and 
 create an optically sectioned image by subtracting the second image from the first image. 
   
     
     
         16 . The system of  claim 15 , wherein each of the first and second cameras are a charge-coupled device (CCD) camera. 
     
     
         17 . The system of  claim 16 , further comprising:
 a first lens;   a second lens spaced-apart from the first lens; and   an emission filter spaced-apart from the second lens,   wherein each of the first lens, the second lens and the emission filter are positioned between the disc and the second camera.   
     
     
         18 . The system of  claim 15 , wherein subtracting the second image from the first image allows nuclear morphology to be more easily discernable to a user than when viewing the first image alone. 
     
     
         19 . The system of  claim 15 , wherein the first and second images are captured simultaneously. 
     
     
         20 . The system of  claim 15 , wherein the first camera is positioned on a first side of the disc and the second camera is positioned on an opposing second side of the disc.