Sequential analysis of biological samples

Abstract

Methods for detecting multiple targets in a biological sample are provided. The methods includes contacting the sample with a first probe; physically binding the first probe to a first target; observing a first signal from the first probe; applying a chemical agent to modify the first signal; contacting the sample with a second probe; physically binding the second probe to a second target; and observing a second signal from the second probe. The methods disclosed herein also provide for multiple iterations of binding, observing, signal modification for deriving information about multiple targets in a single sample. An associated kit and device are also provided.

1 . A device, comprising a sample handling system; a reagent dispensing system; and a signal detection system, wherein the device is operable to detect multiple targets in a biological sample using the method comprising:
 (a) contacting the sample with a first probe to physically bind the probe to a first target;   (b) detecting a first signal from the first probe after the first probe has bound to the first target;   (c) applying a chemical agent to modify the first signal;   (d) contacting the sample with a second probe to physically bind the probe to a second target;   (e) detecting a second signal from the second probe after the second probe has bound to a second target.   
     
     
         2 . The device of  claim 1 , wherein the reagent dispensing system provides for contacting the sample with the first probe, the second probe, and the chemical agent. 
     
     
         3 . The device of  claim 1 , wherein at least one of the sample handling system, reagent dispensing system, or the signal detection system are operable without operator intervention. 
     
     
         4 . The device of  claim 1 , wherein said method further comprises measuring intensity values of the signals. 
     
     
         5 . The device of  claim 4 , wherein said method further comprises comparing the signal intensities of the first and second signals, and ascertaining the relative amounts of the first target and the second target. 
     
     
         6 . The device of  claim 4 , wherein said method further comprises correlating the intensity values of each signal to the amount of its respective target in the biological sample. 
     
     
         7 . The device of  claim 6 , wherein a correlation between the amount of target and the signal intensity is determined using a calibration standard. 
     
     
         8 . The device of  claim 6 , wherein the correlating step is performed using computer-aided means. 
     
     
         9 . The device of  claim 1 , wherein said step of detecting a first signal from the first probe further comprises a step of capturing an image of the biological sample. 
     
     
         10 . The device of  claim 1 , wherein said step of detecting a second signal from the second probe further comprises a step of capturing an image of the biological sample. 
     
     
         11 . The device of  claim 1 , wherein one or more of the detecting steps is performed using computer-aided means. 
     
     
         12 . The device of  claim 1 , wherein the signals are stored in the form of digital images, and said method further comprises conducting computer-aided analysis of the images. 
     
     
         13 . The device of  claim 1 , wherein the signals are stored in the form of digital images, and the images are overlaid using computer-aided superimposition to obtain information of the biological sample. 
     
     
         14 . The device of  claim 1 , wherein the signal detection system includes a microscope connected to an imaging device. 
     
     
         15 . The device of  claim 1 , wherein the signal detection system includes an electron spin resonance (ESR) detection system, a charge coupled device (CCD) detection system, a fluorescent detection system, an electrical detection system, a photographic film detection system, a chemiluminescent detection system, an enzyme detection system, an atomic force microscopy (AFM) detection system, a scanning tunneling microscopy (STM) detection system, an optical detection system, a near field detection system, or a total internal reflection (TIR) detection system. 
     
     
         16 . The device of  claim 1 , wherein one or more of steps (a) to (e) are automated. 
     
     
         17 . The device of  claim 1 , where all of steps (a) to (e) are automated.