Method and apparatus for screening chemical compounds

Abstract

Methods and apparatus for screening large numbers of chemical compounds and performing a wide variety of fluorescent assays, including live cell assays. The methods utilize a laser linescan confocal microscope with high speed, high resolution and multi-wavelength capabilities and real time data-processing. Imaging may be done at video-rates and with use of ultraviolet illumination.

1 - 12 . (canceled) 
     
     
         13 . A system for automated analysis of multiple biological samples comprising:
 an image acquisition unit arranged to acquire a digital image of at least one labeled species in each biological sample;   an image processing unit arranged to perform real-time data analysis of the digital images essentially simultaneous with image acquisition, wherein the real-time data analysis involves:
 detecting image objects in the digital images of the biological samples; and 
 measuring one or more morphological, temporal and/or spatial quantities of detected image objects. 
   
     
     
         14 . The system of  claim 13 , wherein the one or more of: object shape, size, orientation, location, number is measured. 
     
     
         15 . The system of  claim 13 , wherein the image acquisition and the real-time data analysis are performed at video rate. 
     
     
         16 . The system of  claim 15 , wherein the image acquisition and the real-time data analysis are performed at a rate of approximately 30 frames per second to enable registration of a time-dependent response of one or more fluorescently-labeled species. 
     
     
         17 . The system of  claim 13 , wherein the image acquisition and the real-time data analysis are performed prior to storage. 
     
     
         18 . The system of  claim 17 , wherein only the results of the measurements are stored. 
     
     
         19 . The system of  claim 13 , wherein the detection step comprises thresholding. 
     
     
         20 . The system of  claim 13 , wherein multiple target cells are present in the biological samples. 
     
     
         21 . The system of  claim 20 , wherein the acquiring step includes acquiring a first digital image of a first labeled species and a second digital image of a second labeled species. 
     
     
         22 . The system of  claim 13 , wherein the labeled species are cellular components tagged with a fluorescent label or a chromogenic label. 
     
     
         23 . The system of  claim 13 , wherein the detection of image objects is performed using a stack data structure. 
     
     
         24 . The system of  claim 13 , wherein the real-time data analysis comprises a localization analysis algorithm or a translocation analysis algorithm. 
     
     
         25 . The system of  claim 13 , wherein the image acquisition unit is an automated line-scan confocal microscope. 
     
     
         26 . A method of analyzing a biological sample, comprising the steps of:
 (a) providing a biological sample comprised of at least one cell with at least one labeled species;   (b) acquiring images of the labeled species in the biological sample as a function of time;   (c) detecting at least one image object in the image of the labeled species;   (d) averaging, for each detected object, the recorded intensity of the labeled species from each of the images in the time series.   (e) deriving statistical information from the set of averaged intensities for the detected objects.   
     
     
         27 . The method of  claim 26 , wherein the statistical information reported for each object is one or more of rise time, fall time and amplitude.