Systems for transiently dynamic flow cytometer analysis

Abstract

A flow cytometry apparatus and methods to process information incident to particles or cells entrained in a sheath fluid stream allowing assessment, differentiation, assignment, and separation of such particles or cells even at high rates of speed. A first signal processor individually or in combination with at least one additional signal processor for applying compensation transformation on data from a signal. Compensation transformation can involve complex operations on data from at least one signal to compensate for one or numerous operating parameters. Compensated parameters can be returned to the first signal processor for provide information upon which to define and differentiate particles from one another.

1 . A method of flow cytometry analysis, comprising the steps of: 
 establishing a fluid stream;    entraining particles in said fluid stream;    sensing a first occurrence incident to at least one particle;    generating a first signal from said first occurrence incident to said at least one particle;    producing data from said first signal;    sensing at least one additional occurrence incident to said at least one particle;    generating at least one additional signal from said at least one additional occurrence incident to said at least one particle;    producing data from said at least one additional signal;    processing data from said first signal;    processing data from said at least one additional signal;    applying at least one transformation operation to processed data from said first signal;    applying at least one transformation operation to processed data from said at least one additional signal;    compensating at least one parameter shared by said first occurrence and said at least one additional occurrence; and    differentiating said first occurrence from said at least one additional occurrence based upon said at least one compensated parameter.    
     
     
         2 . A method of flow cytometry analysis as described in  claim 1 , wherein said steps of sensing a first occurrence incident to at least one particle and sensing at least one additional occurrence incident to said at least one particle comprise sensing occurrences incident to a single particle.  
     
     
         3 . A method of flow cytometry analysis as described in  claim 1 , wherein said steps of sensing a first occurrence incident to at least one particle and sensing at least one additional occurrence incident to said at least one particle comprise sensing occurrences incident to at least two particles.  
     
     
         4 . A method of flow cytometry analysis as described in  claim 1 , wherein said steps of applying at least one transformation operation to processed data from said first signal and at least one transformation operation to processed data from said at least one additional signal comprises performing complex operations on said processed data from first signal and on said processed data from said at least one additional signal.  
     
     
         5 . A method of flow cytometry analysis as described in  claim 4 , wherein said step of performing complex operations on said processed data from first signal and on said processed data from said at least one additional signal comprises performing algebraic operations.  
     
     
         6 . A method of flow cytometry analysis as described in  claim 5 , wherein performing algebraic operations comprises: 
 applying a parameter compensation transformation to said first signal and to said at least one additional signal;    generating a first compensated signal and at least one additional compensated signal; and    comparing said first compensated signal and said at least one additional compensated signal.    
     
     
         7 . A method of flow cytometry analysis as described in  claim 1 , further comprising the step of using a first signal processor and at least one additional signal processor in said steps of processing said data from said first signal and processing said data from said at least one additional signal.  
     
     
         8 . A flow cytometer analyzer comprising: 
 a fluid stream;    at least one particle entrained in said fluid stream;    a first sensor responsive to said at least one particle entrained in said fluid stream;    at least one signal generator;    data from said at least one signal generator incident to a first occurrence;    data from said at least one signal generator incident to at least one additional occurrence;    a signal processor of said data from said at least one signal generator;    a transformation operation applied to at least a portion of said data from said at least one signal generator incident to said first occurrence;    a transformation operation applied to at least a portion of said data from said at least one signal generator incident to said at least one additional occurrence;    a compensated parameter shared by said first occurrence and by said at least one additional occurrence; and    a particle differentiation element configured to differentiate said first occurrence from said at least one additional occurrence.    
     
     
         9 . A flow cytometer analyzer as described in  claim 8 , wherein said transformation operation applied to at least a portion of said data from said at least one signal generator incident to said first occurrence and to said transformation operation applied to at least a portion of said data from said at least one signal generator incident to said at least one additional occurrence comprises a complex transformation operation.  
     
     
         10 . A flow cytometer analyzer as described in  claim 9 , further comprising at least one additional signal processor.  
     
     
         11 . A flow cytometer analyzer as described in  claim 10 , wherein said at least one additional signal processor performs said complex transformation operation applied to said at least a portion of said data from said at least one signal generator incident to said first occurrence and applied to at least a portion of said data from said at least one signal generator incident to said at least one additional occurrence.  
     
     
         12 . A flow cytometer analyzer as described in  claim 11 , wherein said at least one additional signal processor is a digital signal processor.  
     
     
         13 . A method of flow cytometry analysis, comprising the steps of: 
 establishing a fluid stream;    sensing an occurrence incident to said fluid stream;    generating a signal from said occurrence;    processing said signal using a first signal processor;    processing said signal using at least one additional signal processor in parallel with said first signal processor;    utilizing at least a portion of code by said first signal processor and said at least one additional signal processor;    combining output from said first signal processor and said at least one additional signal processor; and    applying said combined output to classify said occurrence.    
     
     
         14 . A method of flow cytometry analysis as described in  claim 13 , wherein said step of processing said signal using at least one additional signal processor in parallel with said first signal processor comprises processing at least a portion of said signal using said first signal processor and said at least one additional signal processor simultaneously.  
     
     
         15 . A method of flow cytometry analysis as described in  claim 13 , further comprising the steps of: 
 performing compensation transformation on said signal; and    generating a compensated signal.    
     
     
         16 . A method of flow cytometry analysis as described in  claim 15 , wherein said step of performing compensation transformation on said signal comprises compensating a single parameter.  
     
     
         17 . A method of flow cytometry analysis as described in  claim 16 , wherein said step of compensating a single parameter comprises compensating an analog signal.  
     
     
         18 . A method of flow cytometry analysis, comprising the steps of: 
 establishing a fluid stream;    sensing serial occurrences incident to said fluid stream;    generating an analog signal corresponding to said serial occurrences;    performing compensation transformation on said analog signal corresponding to said serial occurrences; and    minimizing variations of said analog signal corresponding to said serial occurrences,    wherein variations are selected from the group consisting of phase and shape.    
     
     
         19 . A method of flow cytometry analysis as described in  claim 18 , wherein said step of performing compensation transformation comprises minimizing shared characteristics.  
     
     
         20 . A method of flow cytometry analysis as described in  claim 19 , wherein said step of minimizing shared characteristics comprises reducing spectrum overlap.