Devices and methods to image objects by time delay integration

Abstract

Devices and methods for automated collection and image analysis are disclosed that enable identification or classification of microscopic objects aligned or deposited on surfaces. Such objects, e.g. detectably labeled rare target cells, are magnetically or non-magnetically immobilized and subjected to Time Delay Integration imaging (TDI). Incorporation of TDI technology into image cytometry analysis, like CellTracks®, makes it possible to image moving objects with very high sensitivity and signal-to-noise ratios. Implementation of TDI camera technology with dual excitation and multispectral imaging of enriched rare cells provides a rapid system for detection, enumeration, differentiation and characterization of imaged rare cells on the basis of size, morphology and immunophenotype.

1 . A method for analytical imaging of target entities, which method comprises: 
 a. obtaining a sample suspected of containing said target entities,    b. magnetically labeling said target entities with magnetic particles that are specific for said target entities,    c. magnetically manipulating said target entities towards a collection surface in a sample chamber,    d. moving said sample chamber on a scanning stage while illuminating said collected target entities,    e. detecting emitted light from said collected target entities, and    f. assessing characteristics of collected target entities from a group consisting of intensity, color, morphology, and combinations thereof.    
     
     
         2 . The method of  claim 1 , further comprising: 
 a. detecting emitted light whereby sequential sub-images of said collected target entities are collected, and    b. re-combining said sub-images to construct a complete image of said collected target entities.    
     
     
         3 . The method of  claim 1 , in which detecting emitted light is by TDI.  
     
     
         4 . The method of  claim 1 , in which said illumination step further comprises the use of multiple wavelength light sources  
     
     
         5 . The method of  claim 1 , in which said illuminating is by excitation with a laser.  
     
     
         6 . The method of  claim 4 , in which said excitation is with a blue and green laser.  
     
     
         7 . The method of  claim 1 , in which said detecting is by fluorescent emission.  
     
     
         8 . The method of  claim 1  wherein said emitted light is from a group consisiting of green fluorescent emitted light, blue fluorescent emitted light, brightfield light, and combinations thereof.  
     
     
         9 . The method of  claim 1 , in which said target entities are from a group consisting of epithelial cells, endothelial cells, tumor cells, cell debris, and combinations thereof.  
     
     
         10 . The method of  claim 1 , in which said magnetic labels are colloidal magnetic particles specific for an antigenic site on the target entity.  
     
     
         11 . The method of  claim 10 , in which said colloidal magnetic particles are specific for the Epithelial Cell Adhesion Molecule (EpCAM).  
     
     
         12 . The method of  claim 1 , in which said collection surface comprises parallel Nickel lines on a glass substrate.  
     
     
         13 . An improved apparatus for analytical imaging of target entities having a sample chamber with a collection surface, an arragnement of magnets cabable of manipulating magnetically labeled target entities towards said collection surface, at least one light source, and a computer, whereby said improvement comprises: 
 a. a scanning stage wit said sample chamber affixed for controlled speed and positioning, and    b. a TDI camera.    
     
     
         14 . The apparatus of  claim 13 , in which said collection surface comprises Nickel lines on a glass substrate.  
     
     
         15 . The apparatus of  claim 13 , in which said light source is a laser.