System and method for automatic color segmentation and minimum significant response for measurement of fractional localized intensity of cellular compartments

Abstract

A system, a method, and a programmed device for measurement of translocational activity among cellular compartments process magnified images of cellular material exposed to an agent by segmenting and compartmentalizing the images and then measuring fractional localized intensity of two or more compartments in the segmented, compartmentalized image. The measured fractional localized intensities are compared to determine translocation of cellular material among the measured components caused by the agent.

1 . A system for measuring cell activity represented in an image obtained with an automated microscopy platform the image including cells treated with an agent that causes translocation of cellular material, the system comprising:
 means for segmenting cellular components in the image;   means for separating segmented components of overlapping cells in the image; and   means for determining translocation of cellular material between cellular compartments in the segmented, separated image based upon a ratio of a first intensity of the cellular material in a first cellular compartment to a sum of at least the first intensity and a second intensity of the cellular material in a second cellular compartment.   
     
     
         2 . The system of  claim 1 , wherein the means for segmenting includes means for enhancing contrast between the first cellular compartments and the background of the image, and between the second cellular compartments and the background of the image. 
     
     
         3 . The system of  claim 1 , wherein the means for separating includes means for tessellating the image. 
     
     
         4 . The system of  claim 1 , wherein the means for determining includes:
 means for measuring a fractionalized local intensity of the first cellular material;   means for measuring a fractionalized local intensity of the second cellular material; and,   means for comparing the fractionalized local intensities of the first and second cellular materials.   
     
     
         6 . The system of  claim 4 , wherein:
 the means for segmenting includes means for enhancing contrast between the first cellular compartments and the background of the image, and between the second cellular compartments and the background of the image; and,   the means for separating includes means for tessellating the image.   
     
     
         7 . The system of  claim 1 , wherein the means for determining includes:
 means for measuring the first intensity of the cellular material in the first cellular compartment a and for measuring the second intensity of the cellular material in the second cellular compartment b; and,   means for determining a fractional localized intensity F a  of the cellular material in the first cellular compartment by a ratio of the first intensity to a sum of at least the first and second intensities, according to:   
       
         
           
             
               
                 F 
                 a 
               
               = 
               
                 
                   
                     ∑ 
                     
                       
                         ( 
                         
                           x 
                           , 
                           y 
                         
                         ) 
                       
                       ∈ 
                       a 
                     
                   
                    
                   
                     I 
                      
                     
                       ( 
                       
                         x 
                         , 
                         y 
                       
                       ) 
                     
                   
                 
                 
                   
                     ∑ 
                     
                       
                         
                           ( 
                           
                             x 
                             , 
                             y 
                           
                           ) 
                         
                         ∈ 
                         a 
                       
                       , 
                       b 
                     
                   
                    
                   
                     I 
                      
                     
                       ( 
                       
                         x 
                         , 
                         y 
                       
                       ) 
                     
                   
                 
               
             
           
         
         where I(x, y), the emission intensity at a pixel location (x, y) in a cellular compartment, is determined according to: 
       
       
         
           
             
               
                 I 
                  
                 
                   ( 
                   
                     x 
                     , 
                     y 
                   
                   ) 
                 
               
               = 
               
                 
                   ∫ 
                   0 
                   z 
                 
                  
                 
                   
                     I 
                     0 
                   
                    
                   Q 
                    
                   
                       
                   
                    
                   ɛ 
                    
                   
                       
                   
                    
                   u 
                    
                   
                      
                     z 
                   
                 
               
             
           
         
         in which I 0  is an incident excitation intensity, Q is a quantum yield, ε is an extinction coefficient, u is a local fluorophore concentration at pixel location (x, y), and z is the thickness of a column of cellular material at pixel location (x, y). 
       
     
     
         8 . The system of  claim 7 , wherein segmenting includes enhancing contrast between first cellular compartments including the first cellular compartment and the background of the image, and between second cellular compartments including the second cellular compartment and the background of the image. 
     
     
         9 . The system of  claim 8 , wherein separating includes tessellating the image. 
     
     
         10 . The system of  claim 7 , wherein separating includes tessellating the image.