Luminescence detecting apparatuses and methods

Abstract

A luminescence detecting apparatus and method for analyzing luminescent samples is disclosed. Luminescent samples are placed in a plurality of sample wells in a tray, and the tray is placed in a visible-light impervious chamber containing a charge coupled device camera. The samples may be injected in the wells, and the samples may be injected with buffers and reagents, by an injector. In the chamber, light from the luminescent samples pass through a collimator, a Fresnel field lens, a filter, and a camera lens, whereupon a focused image is created by the optics on the charge-coupled device (CCD) camera. The use of a Fresnel field lens, in combination with a collimator and filter, reduces crosstalk between samples below the level attainable by the prior art. Preferred embodiments of the luminescence detecting apparatus and method disclosed include central processing control of all operations, multiple wavelength filter wheel, and robot handling of samples and reagents. Preferred embodiments of processing software integrated with the invention include elements for mechanical alignment, outlier shaving, edge detection and masking, manipulation of multiple integration times to expand the dynamic range, crosstalk correction, dark subtraction interpolation and drift correction, multi-component analysis applications specifically tailored for luminescence, and uniformity correction.

1 - 13 . (canceled) 
     
     
         14 . A luminescence detecting apparatus, comprising:
 a sample chamber configured to receive a plurality of sampling areas containing respective luminescent samples;   a photosensitive detector;   an optical system configured to produce a sample image of the sampling areas, comprising:
 a substrate comprising a two-dimensional array of openings and positioned during use between the plurality of sampling areas and the photosensitive detector, the plurality of sampling areas aligned during use to the array of openings, the array of openings configured to simultaneously pass emissions from at least some of the sample areas to the photosensitive detector and to block some of the emissions from being received by the photosensitive detector; and 
 a lens disposed along an optical path between the two-dimensional array of openings and the photosensitive detector; and 
   a processor comprising instructions to a subtract a dark image from the sample image;   wherein the array of openings, the lens, and the processor are together configured to refine values of the sample image.   
     
     
         15 . The apparatus of  claim 14 , wherein the plurality of sampling areas comprises an array of sample wells. 
     
     
         16 . The apparatus of  claim 14 , further comprising a defogger configured to prevent condensation. 
     
     
         17 . The apparatus of  claim 14 , wherein the photosensitive detector is configured to sequentially detect light at different wavelengths emitted from the plurality of sampling areas. 
     
     
         18 . The apparatus of  claim 14 , further comprising a plurality of luminescent samples disposed within respective ones of the plurality of sample areas during use. 
     
     
         19 . The apparatus of  claim 18 , wherein each of the plurality of luminescent samples is one of a bioluminescent material or a chemiluminescent material. 
     
     
         20 . The apparatus of  claim 14 , further comprising a plate, wherein plurality of sampling areas are disposed on the plate. 
     
     
         21 . The apparatus of  claim 14 , further comprising a filter disposed along the optical path between the two-dimensional array of openings and the detector, the filter configured to permit passage of a specific wavelength of light emitted from the sample areas. 
     
     
         22 . The apparatus of  claim 14 , wherein the two-dimensional array of openings is in close proximity to the plurality of sampling areas during use, so as to enhance restriction of stray light from the sampling areas. 
     
     
         23 . The apparatus of  claim 14 , wherein the lens comprises at least one of a Fresnel lens, an aspheric lens, or a molded plastic lens. 
     
     
         24 . The apparatus of  claim 14 , wherein the photosensitive detector comprises a plurality of photosensitive detector elements. 
     
     
         25 . The apparatus of  claim 14 , wherein the photosensitive detector comprises a charge coupled device. 
     
     
         26 . The apparatus of  claim 25 , wherein:
 the lens is a primary lens and the apparatus further comprises a camera lens disposed between the primary lens and the charge coupled device; and   the camera lens is configured to simultaneously image the luminescent samples of more than one of sampling areas.   
     
     
         27 . The apparatus of  claim 14 , wherein the sample chamber comprises a bottom surface and the substrate is positioned during use above the sample chamber. 
     
     
         28 . The apparatus of  claim 14 , wherein the plurality of sampling areas are in simultaneous optical communication with the photosensitive detector, and wherein the photosensitive detector and the two-dimensional array of openings together are configured to simultaneously detect the plurality of respective luminescent samples contained in the plurality of sampling areas. 
     
     
         29 . The apparatus of  claim 14 , wherein the sample chamber comprises a bottom surface and the substrate is positioned during use above the sample chamber. 
     
     
         30 . A method for analyzing a plurality of luminescent samples, comprising the steps of:
 providing a plurality of luminescent samples disposed in a respective plurality of sampling areas;   providing a photosensitive detector;   providing an optical system configured to produce a sample image of the sampling areas, the optical system comprising:
 a substrate comprising a two-dimensional array of openings and positioned during use between the plurality of sampling areas and the photosensitive detector, the plurality of sampling areas aligned during use to the array of openings, the array of openings configured to simultaneously pass emissions from at least some of the sample areas to the photosensitive detector and to block some of the emissions from being received by the photosensitive detector; and 
 a first lens disposed along an optical path between the two-dimensional array of openings and the photosensitive detector; and 
   placing the plurality of sampling areas in optical communication with the optical system;   passing emissions from at least some of the sampling areas through the two-dimensional array of openings to respective ones of the photosensitive detector;   using the substrate to prevent some of the emissions from the at least some of the sampling areas from being received by the photosensitive detector;   using the photosensitive detector:
 producing a dark image; and 
 producing a sample image 
   using the array of openings, the lens, and the dark image to produce refined values of the sample image.   
     
     
         31 . The apparatus of  claim 30 , wherein the photosensitive detector comprises a plurality of photosensitive detector elements. 
     
     
         32 . The apparatus of  claim 30 , wherein the plurality of sampling areas comprises an array of sample wells. 
     
     
         33 . The method of  claim 30 , further comprising positioning a second lens disposed along the optical path at a location between the first lens and the photosensitive detector.