Systems and methods for cell culture device interconnection and fluidic device interconnection

Abstract

Herein is described kinetic assay, in which individual binding events are detected and monitored during sample incubation. This method uses interferometric reflectance imaging to detect thousands of individual binding events across a multiplex solid phase sensor with a large area. A dynamic tracking procedure is used to measure the duration of each event. From this, the total rates of binding and de-binding as well as the distribution of binding event durations are determined. Systems and components for performing the kinetic assay are also described.

What is claimed is: 
     
         1 . A method for detecting at least one particle, the method comprising:
 providing a surface including a capture agent,   providing a camera for generating an image of the surface,   providing a computer, including a memory and a processor, connected to the camera;   flowing a solution containing particles over the surface, thereby contacting the particles with the capture agent,   collecting a plurality of particle-images of the surface using the camera and storing said particle-images in the memory,   analyzing the plurality of particle-images using the processor to produce a catalog of particles and time associated data for each particle in the catalog and storing said catalog in the memory,   eliminating a particle from the catalog in the memory if the time associated data of said particle does not span a time greater than a set time increment,   outputting data from the catalog as the detection of the at least one particle.   
     
     
         2 . The method of  claim 1 , further comprising combining a first particle and a second particle in the catalog, so that said combining identifies the first and second particle as a single particle in the catalog, if said first particle is listed in a first particle-image sequence in the catalog and then not listed is a subsequent second particle-image sequence of the catalog, and said second particle is listed in a third particle-image sequence of the catalog, and wherein the time spanned by the second particle-image sequence in the catalog corresponds to less than the time increment, and wherein said combining includes assigning the time spanning the first particle-image sequence in the catalog, the second particle-image sequence in the catalog and the third particle-image sequence in the catalog to the first particle in the catalog. 
     
     
         3 . The method as in  claim 1 , further comprising providing a user interface and outputting said data comprises providing catalog-representative data to the user interface. 
     
     
         4 . The method of  claim 3 , wherein said catalog-representative data is in the form of a text data, image data, or graphed data. 
     
     
         5 . The method of  claim 1 , wherein the surface is a component in an interferometric reflectance imaging sensor (IRIS) system comprising an objective lens for illuminating a detection region of a cartridge and collecting reflected light from the detection region, wherein the detection region includes the surface. 
     
     
         6 . The method as in  claim 1 , wherein said capture agent is an antibody, a protein, a peptide an oligonucleotide, a complexing ligand, a single stranded DNA or RNA, a haptan, or a polymer. 
     
     
         7 . The method as in  claim 1 , wherein said set time increment is between one second and one hour. 
     
     
         8 . The method of  claim 1 , wherein said particles comprise a nanoparticle functionalized with a target, said capture agent having a high affinity for the target. 
     
     
         9 . The method of  claim 8 , wherein the target is selected from the group consisting of a small molecule, a polymer, an antibody, a haptan, an oligonucleotide, a single stranded DNA or RNA, a protein and a peptide. 
     
     
         10 . The method of  claim 1 , wherein the particle comprises a nanoparticle. 
     
     
         11 . The method of  claim 1 , wherein the particle is a gold nano-particle. 
     
     
         12 . The method of  claim 1 , wherein the particle is a virus. 
     
     
         13 . An apparatus for measuring particles comprising;
 a surface including a capture agent,   a camera for imaging the surface,   a user interface for visualization of particle data,   a system for flowing a solution of particles over and in contact with the surface, and   a computer including a memory and a processor connected to the camera and the user interface;   wherein the camera is configured to provide a plurality of particle-images of the surface and storing said particle-images in the memory,   wherein said analyzing comprises using the processor to produce a catalog of particles and time associated data for each particle in the catalog, and storing said catalog in the memory,   wherein analyzing further comprises eliminating any particle from the catalog in the memory if said the time associated data associated with said particle does not span a time greater than a set time increment, and   wherein the computer is configured to send representative data to the user interface for visualization of catalog-representative data.   
     
     
         14 . The apparatus according to  claim 13 , wherein analyzing further comprises combining a first particle and a second particle in the catalog, so that said combining identifies the first and second particle as a single particle in the catalog, if said first particle is listed in a first particle-image sequence in the catalog and then not listed is a subsequent second particle-image sequence in the catalog, and said second particle is listed in a third particle-image sequence in the catalog, and wherein the time spanned by the second particle-image sequence in the catalog corresponds to less than the time increment, and wherein said combining includes assigning the time spanning the first particle-image sequence in the catalog, the second particle-image sequence in the catalog and the third particle-image sequence in the catalog to the first particle in the catalog. 
     
     
         15 . The apparatus according to  claim 13 , wherein the surface is a component in an interferometric reflectance imaging sensor (IRIS) system comprising an objective lens for illuminating a detection region of a cartridge and collecting reflected light from the detection region, wherein the detection region includes the surface. 
     
     
         16 . The apparatus according to  claim 15 , wherein the system for flowing a solution of particles includes an inlet and an outlet to the cartridge, wherein the solution flows through the cartridge from the inlet to the outlet. 
     
     
         17 . The apparatus of  claim 13 , further comprising a stage for placement of the surface thereupon and configured for relative movement of the surface to the camera. 
     
     
         18 . A kinetic assay for the detection of single binding events, comprising 
 flowing a solution containing particles over a capture agent functionalized surface,   collecting a plurality of particle-images of the surface using a camera and storing said particles-images in a computer memory,   analyzing the plurality of particle-images using a processor to produce a catalog of particles and time associated data for each particle in the catalog,   eliminating a particle from the catalog if said particle is not found in sub-sequence of particle-images corresponding to a time lapse greater than a set time increment,   combining a first particle and a second particle in the catalog, so that said combining identifies the first and second particle as a single particle in the catalog, if said first particle is listed in a first particle-image sequence of the catalog and then not listed is a subsequent second particle-image sequence of the catalog, and said second particle is listed in a third particle-image sequence of the catalog, and wherein the time spanned by the second particle-image sequence in the catalog corresponds to less than the time increment, and wherein said combining includes assigning the time spanning the first particle-image sequence in the catalog, the second particle-image sequence in the catalog and the third particle-image sequence in the catalog to the first particle in the catalog,   wherein each particle in the catalog corresponds to a binding event.   
     
     
         19 . The assay as in  claim 18 , wherein the binding event detects specific binding of the particle to the surface when the concentration of the particle is below the critical concentration. 
     
     
         20 . The assay as in  claim 18 , wherein the assay does not use a chemical amplification reaction.