Apparatus and method for interleaving detection of fluorescence and luminescence

Abstract

An apparatus is provided that is capable of interleaving detection of fluorescence and luminescence signals emitted from a plurality of samples. The apparatus is suitable for analysis of samples containing single cells or tissues up to and including living organisms. It contains an optical assembly or “sandwich” for producing a spectrally pure and spatially disperse light source for illuminating the sample. The invention also provides a plurality of optical sandwiches that can be variously geometrically arranged and their intensities programmed to create spatially uniform illumination over a large sample. The invention further provides an apparatus having at least one of the optical sandwich and a detector system capable of interleaving detection of fluorescent and luminescent signals when a suitable sample is illuminated by the light source of the optical sandwich. Methods for preparing samples and using the sandwiches, arrays and apparatus, are further provided by this invention. A method for interleaving detection of fluorescent and luminescent signals emitted from a plurality of samples is disclosed.

1 . A method for detecting a signal emitted from a fluorescently and luminescently labeled sample, comprising illuminating the sample with a spectrally pure and spatially dispersed illumination from an excitation light source and detecting any fluorescent or luminescent signal emitted from the sample with a detector system, wherein the excitation light source for illuminating the sample is automatically turned off or shuttered during interleaved luminescence imaging and wherein the detector system comprises: (i) a detector lens (ii) an emission filter and (iii) a single sensor capable of detecting a two dimensional image plane. 
   
   
       2 . The method of  claim 1 , wherein the sample comprises two different fluorescent labels. 
   
   
       3 . The method of  claim 2 , wherein the plurality of samples express or are labeled with more than one luminescent label. 
   
   
       4 . The method of  claim 1 , wherein the plurality of samples express or are labeled with one or more fluorescent label and with one or more luminescent label. 
   
   
       5 . The method of  claim 1 , wherein the luminescent label is a bioluminescent label or a chemical luminescent label. 
   
   
       6 . The method of  claim 2 , wherein the luminescent label is a bioluminescent label or a chemical luminescent label. 
   
   
       7 . The method of  claim 3 , wherein the luminescent label is a bioluminescent label or a chemical luminescent label. 
   
   
       8 . The method of  claim 4 , wherein the luminescent label is a bioluminescent label or a chemical luminescent label. 
   
   
       9 . A method comprising interleaving detection of fluorescent and luminescent signals emitted from a plurality of samples, comprising illuminating the plurality of samples with a spectrally pure and spatially dispersed illumination from an excitation light source and detecting any fluorescent or luminescent signals emitted from the plurality of samples with a detector system comprising: (i) a detector lens, (ii) an emission filter, and (iii) a single sensor capable of detecting a two dimensional image plane, wherein the excitation light source for illuminating the plurality of samples is automatically turned off or shuttered during interleaved luminescence imaging. 
   
   
       10 . The method of  claim 9 , wherein the sample comprises two different fluorescent labels. 
   
   
       11 . The method of  claim 10 , wherein the plurality of samples express or are labeled with more than one luminescent label. 
   
   
       12 . The method of  claim 9 , wherein the plurality of samples express or are labeled with one or more fluorescent label and with one or more luminescent label. 
   
   
       13 . The method of  claim 9 , wherein the luminescent label is a bioluminescent label or a chemical luminescent label. 
   
   
       14 . The method of  claim 10 , wherein the luminescent label is a bioluminescent label or a chemical luminescent label. 
   
   
       15 . The method of  claim 11 , wherein the luminescent label is a bioluminescent label or a chemical luminescent label. 
   
   
       16 . The method of  claim 12 , wherein the luminescent label is a bioluminescent label or a chemical luminescent label. 
   
   
       17 . A method for interleaving detection of fluorescent and luminescent signals emitted from a plurality of samples, comprising detecting fluorescent and luminescent signals produced by the plurality of samples, wherein the fluorescence excitation light source is automatically turned off or shuttered during interleaved luminescence imaging.