Visual-servoing optical microscopy

Abstract

The present invention provides methods and devices for the knowledge-based discovery and optimization of differences between cell types. In particular, the present invention provides visual servoing optical microscopy, as well as analysis methods. The present invention provides means for the close monitoring of hundreds of individual, living cells over time; quantification of dynamic physiological responses in multiple channels; real-time digital image segmentation and analysis; intelligent, repetitive computer-applied cell stress and cell stimulation; and the ability to return to the same field of cells for long-term studies and observation. The present invention further provides means to optimize culture conditions for specific subpopulations of cells.

1 . (canceled) 
     
     
         2 . A detection device for monitoring cellular responses in a living cell population,
 the detection device comprising a visual-servoing optical microscope and one or more stimulating devices,   wherein the visual-servoing optical microscope is configured to obtain cellular image data from a living cell population and detect differences in the living cell population,   wherein the one or more stimulating devices is configured to independently and automatically stimulate the living cell population in response to detected differences in the living cell population monitored with the visual-servoing optical microscope.   
     
     
         3 . The detection device of  claim 2 , wherein the obtained cellular image data comprises one or more sets of data selected from the group consisting of data attributable to specific types of living cells, data attributable to specific living cells, data attributable to subcellular components of specific types of living cells, and data attributable to subcellular components of specific living cells. 
     
     
         4 . The detection device of  claim 2 , wherein the detected differences in a living cell population comprise detected differences over a series of time points. 
     
     
         5 . The detection device of  claim 2 , wherein any portion of the detected differences in a living cell population comprises one or more detected differences selected from the group consisting of detected differences attributable to specific types of living cells, detected differences attributable to specific living cells, detected differences attributable to subcellular components of specific types of living cells, and detected differences attributable to subcellular components of specific living cells. 
     
     
         6 . The detection device of  claim 2 , wherein any portion of the detected differences in a living cell population comprises one or more detected differences selected from the group consisting of detected differences attributable to specific types of living cells detected over one or more time points, detected differences attributable to specific living cells detected over one or more time points, detected differences attributable to subcellular components of specific types of living cells detected over one or more time points, and detected differences attributable to subcellular components of specific living cells detected over one or more time points. 
     
     
         7 . The detection device of  claim 2 , wherein any portion of the detected differences in a living cell population comprises detected differences between individual living cells and/or detected differences between the subcellular components of individual living cells. 
     
     
         8 . The detection device of  claim 2 , wherein the detected differences in the living cell population comprise detected differences between different types of living cells and/or detected differences between the subcellular components of different types of living cells. 
     
     
         9 . The detection device of  claim 2 , wherein the detected differences in the living cell population comprise detected differences between different types of living cells detected over one or more time points, and/or detected differences between the subcellular components of different types of living cells detected over one or more time points. 
     
     
         10 . The detection device of  claim 2 , wherein one or more living cells within the living cell population are configured to report changes in physiological state. 
     
     
         11 . The detection device of  claim 2 , wherein said device further comprises a local and/or remote knowledge-base configured to interact with said visual-servoing optical microscope. 
     
     
         12 . The detection device of  claim 2 , wherein the visual-servoing optical microscope comprises a specimen vessel, wherein said specimen vessel has an internal frame of reference. 
     
     
         13 . The detection device of  claim 2 , wherein the detected differences in a living cell population comprises detected differences between the living cell population and a reference database. 
     
     
         14 . The detection device of  claim 2 , wherein the one or more stimulating devices are syringes. 
     
     
         15 . A system for recording cellular responses in a living cell population comprising a processor, living cell population, and a detection device comprising a visual-servoing optical microscope and one or more stimulating devices configured to apply a plurality of stimuli to said living cell population,
 wherein the visual-servoing optical microscope is configured to monitor the living cells, obtain cellular image data, and provide the obtained cellular image data to the processor,   wherein the processor is configured to define the obtained cellular image data,   wherein the one or more stimulating devices configured to apply a plurality of stimuli to said living cell population are configured to independently and automatically stimulate the living cell population in response to defined obtained cellular image data obtained with the visual-servoing optical microscope.   
     
     
         16 . The system device of  claim 15 , wherein the obtained cellular image data comprises at least one or more selected from the group consisting of the morphometry of cells or subcellular components within the living cell population, the color of cells or subcellular components within the living cell population, and/or the motion of cells or subcellular components within the living cell population. 
     
     
         17 . The system device of  claim 15 , wherein defined cellular image data comprises defining of the presence or absence of differences in a living cell population over a series of time points for the living cell population. 
     
     
         18 . The system device of  claim 15 , wherein defined cellular image data comprises defining of detected differences in the living cell population over a series of time points. 
     
     
         19 . The system device of  claim 18 , wherein any portion of the detected differences in the living cell population comprises one or more detected differences selected from the group consisting of detected differences attributable to specific types of living cells, detected differences attributable to specific living cells, detected differences attributable to subcellular components of specific types of living cells, and detected differences attributable to subcellular components of specific living cells. 
     
     
         20 . The system device of  claim 18 , wherein any portion of the detected differences in the living cell population comprises one or more detected differences selected from the group consisting of detected differences attributable to specific types of living cells detected over one or more time points, detected differences attributable to specific living cells detected over one or more time points, detected differences attributable to subcellular components of specific types of living cells detected over one or more time points, and detected differences attributable to subcellular components of specific living cells detected over one or more time points. 
     
     
         21 . The system device of  claim 18 , wherein any portion of the detected differences in the living cell population comprises detected differences between individual living cells and/or detected differences between the subcellular components of individual living cells. 
     
     
         22 . The system device of  claim 18 , wherein any portion of the detected differences in the living cell population comprises detected differences between different types of living cells and/or detected differences between the subcellular components of different types of living cells. 
     
     
         23 . The system device of  claim 18 , wherein any portion of the detected differences in the living cell population comprises detected differences between different types of living cells detected over one or more time points, and/or detected differences between the subcellular components of different types of living cells detected over one or more time points. 
     
     
         24 . The system device of  claim 18 , wherein any portion of the detected differences in the living cell population comprises detected differences between the living cell population and a reference database. 
     
     
         25 . The system of  claim 15 , wherein the visual-servoing optical microscope comprises a specimen vessel, wherein said specimen vessel has an internal frame of reference. 
     
     
         26 . The system device of  claim 15 , wherein defined cellular image data comprises defining of the presence or absence of differences in a living cell population in comparison with a reference database. 
     
     
         27 . The system device of  claim 15 , wherein the one or more stimulating devices configured to apply a plurality of stimuli to said living cell population are syringes. 
     
     
         28 . The system device of  claim 15 , wherein one or more living cells within the living cell population is configured to report changes in physiological state. 
     
     
         29 . The system device of  claim 15 , wherein said system further comprises a local and/or remote knowledge-base configured to interact with said visual-servoing optical microscope.