Method and apparatus for analyzing individual cells or particulates using fluorescent quenching and/or bleaching

Abstract

A method for analyzing a blood sample is provided that includes the steps of: providing a blood sample having one or more of each first and second constituents; admixing a colorant with the sample, which colorant is operative to cause the first constituents and second constituents to fluoresce and absorb light; illuminating at least a portion of the sample; e) imaging a portion of the sample; determining a fluorescence value for each the first constituents and second constituents; determining an optical density value for each of the first constituents and second constituents; and identifying the first constituents and the second constituents using the determined fluorescence and optical density values.

What is claimed is: 
     
         1 . A method for identifying constituents within biologic fluid sample, which sample has at least one first constituent and at least one second constituent, and which second constituent is a different type than that of the first constituent, and which sample is mixed with a colorant, the method comprising:
 using a processor to execute stored instructions, which instructions cause the processor to:
 control a sample illuminator to illuminate at least a portion of the sample containing the first constituent and the second constituent with light at one or more wavelengths that cause the colorant to fluoresce, and with light at one or more wavelengths that are absorbed by the colorant; 
 control an imaging device to image the at least a portion of the sample, including producing first image signals indicative of fluorescent emissions from the illuminated at least a portion of the sample, and including producing second image signals indicative of the optical density of the illuminated at least a portion of the sample; 
 determine at least one first fluorescent emissions value associated with the first constituent using the first image signals and at least one second fluorescent emissions value associated with the second constituent using the first image signals; 
 determine at least one first optical density value associated with the first constituent using the second image signals and at least one second optical density value associated with the second constituent using the second image signals; and 
 identify at least one of the first constituent and the second constituent using the determined first and second fluorescent emission values and the determined first and second optical density values. 
   
     
     
         2 . The method of  claim 1 , wherein the first constituent and the second constituent are selected from the group consisting of eosinophils, neutrophils, and basophils. 
     
     
         3 . The method of  claim 1 , wherein the first and second constituents are selected from the group consisting of white blood cells and particulates. 
     
     
         4 . The method of  claim 3 , wherein the first and second constituents are each types of white blood cells. 
     
     
         5 . The method of  claim 1 , wherein the first and second constituents are each types of particulates. 
     
     
         6 . The method of  claim 1 , wherein the first and second constituents are selected from a group consisting of platelets and hematoparasites. 
     
     
         7 . An apparatus for analyzing a biologic fluid sample, which sample has at least one first constituent and at least one second constituent, and which second constituent is a different type than that of the first constituent, and which sample is mixed with a colorant, comprising:
 a light source;   at least one light detector;   a processor that executes stored instructions that:
 control the light source to illuminate at least a portion of the sample containing the first constituent and the second constituent with light at one or more first wavelengths that cause the colorant to fluoresce, and with light at one or more second wavelengths that are absorbed by the colorant; 
 control the at least one light detector to detect light emitted from the at least a portion of the sample, and to produce first image signals indicative of the emitted light from the sample, which emitted light is a fluorescent emission resulting from the illumination of the at least a portion of the sample with the one or more first wavelengths, and to detect transmitted light, which transmitted light is the one or more second wavelengths of light from the light source that has passed through the at least a portion of the sample, and to produce second image signals indicative of the transmitted light; 
 determine at least one first fluorescent emissions value associated with the first constituent using the first image signals and at least one second fluorescent emissions value associated with the second constituent using the first image signals; 
 determine at least one first optical density value associated with the first constituent using the second image signals and at least one second optical density value associated with the second constituent using the second image signals; and 
 identify at least one of the first constituent and the second constituent using the determined first and second fluorescent emission values and the determined first and second optical density values. 
   
     
     
         8 . The apparatus of  claim 7 , wherein the first and second constituents are selected from the group consisting of white blood cells and particulates. 
     
     
         9 . The apparatus of  claim 7 , wherein the first and second constituents are each types of white blood cells. 
     
     
         10 . The apparatus of  claim 9 , wherein the first constituents and the second constituents are selected from the group consisting of eosinophils, neutrophils, and basophils. 
     
     
         11 . The apparatus of  claim 7 , wherein the first and second constituents are each types of particulates. 
     
     
         12 . The apparatus of  claim 7 , wherein the first and second constituents are selected from a group consisting of platelets and hematoparasites.