Darkfield imaging system and methods for automated screening of calls

Abstract

The disclosure is directed at a system and method for darkfield imaging system and method for automated cell screening of cells. The system and method acquires multi- or hyperspectral digital darkfield images of cells and then processes the images to obtain measurements which can then be supplied or displayed to a user to analyze.

1 - 20 . (canceled) 
     
     
         21 . A system for screening at least one cell comprising:
 image acquiring apparatus configured to acquire one or more images of the at least one cell in color at one or more of wavelengths and wavelength bands, via one or more of transmitted illumination and reflected illumination, the one or more of the transmitted illumination and the reflected illumination comprising one or more of oblique illumination and darkfield illumination; and,   a processor, in communication with the image acquiring apparatus, the processor configured to:
 process chrominance data in the one or more images using one or more color space models to segment and measure one or more of specific cells and cell groups within the one or more images of the at least one cell, the chrominance data comprising native color information of the at least one cell. 
   
     
     
         22 . The system of  claim 21 , wherein the processor is configured to process the chrominance data in the one or more images using the one or more color space models to obtain measurements relating to the at least one cell. 
     
     
         23 . The system of  claim 21 , further comprising a microscope comprising a stage, the image acquiring apparatus located at an end of the microscope away from the stage. 
     
     
         24 . The system of  claim 23 , wherein the stage comprises a motorized stage. 
     
     
         25 . The system of  claim 21  further comprising: a test substance delivery system configured to deliver test substances at one or more specified locations to the at least one cell. 
     
     
         26 . The system of  claim 25 , wherein the test substance delivery system is further configured to dispense different test substances at different locations. 
     
     
         27 . The system of  claim 21 , further comprising an imaging chamber configured to contain the at least one cell. 
     
     
         28 . The system of  claim 27 , wherein the imaging chamber comprises one or more of a glass slide, a multi-well plate, a tissue microarray, a chip or a microfluidic device. 
     
     
         29 . A method for screening at least one cell comprising:
 acquiring one or more images of the at least one cell in color at one or more of specified wavelengths and wavelength bands, via one or more of transmitted illumination and reflected illumination, the one or more of the transmitted illumination and the reflected illumination comprising one or more of oblique illumination and darkfield illumination, using an image acquiring apparatus; and,   processing, using a processor, chrominance data in the one or more images using one or more color space models to segment and measure one or more of specific cells and cell groups within the one or more images, the chrominance data comprising native color information of the at least one cell.   
     
     
         30 . The method of  claim 29 , wherein the processing of the chrominance data comprises using a luminance-chrominance color space model to obtain the measurements relating to the at least one cell. 
     
     
         31 . The method of  claim 30 , wherein the processing of the chrominance data further comprises: inverting the one or more images in luminance-chrominance color space to obtain one or more of an inverted luminance image, an inverted chrominance image and a combined inverted chrominance and luminance image. 
     
     
         32 . The method of  claim 30 , wherein the processing of the chrominance data further comprises: using one or more additional color space models to process the chrominance data in combination with the luminance-chrominance color space model. 
     
     
         33 . The method of  claim 29 , wherein the processing of the chrominance data comprises weighting the information obtained from the measurements in order of statistical importance. 
     
     
         34 . The method of  claim 29  further comprising:
 acquiring one or more color images of entities that one or more of: are located outside the at least one cell; have entered the at least one cell; and have invaded the at least one cell; and 
 processing chrominance data in the one or more color images using one or more color space models to obtain respective measurements relating to the entities. 
 
     
     
         35 . The method of  claim 34 , wherein the entities comprise one or more of viruses, prokaryotes, foreign molecules and natural endogenous components of the at least one cell. 
     
     
         36 . The method of  claim 34 , wherein the respective measurements provide further information about one or more of effects of test substances on the entities and time-dependent changes in the entities. 
     
     
         37 . The method of  claim 29 , wherein the at least one cell is selected from a group consisting of: living cells, dead cells, prokaryotic cells, eukaryotic cells, immortal cells, primary cells, and cells in a tissue. 
     
     
         38 . The method of  claim 29 , wherein the measurements provide information about one or more of effects of test substances and time-dependent changes in one or more of the following states of cells: cell viability, spreading, differentiation, transformation, aging, necrosis, apoptosis, morphological shape, relationships of the cells with each other in a population of the cells, infection or invasion of the cells by exogenous agents, receptor activation and toxic effects of test substances on the cells. 
     
     
         39 . The method of  claim 29 , wherein the acquiring comprises: collecting the one or more images at different focal planes to obtain a 3D image of the at least one cell. 
     
     
         40 . The method of  claim 29 , wherein the acquiring comprises: repeatedly imaging the at least one cell to analyze time-dependent changes in the measurements.