Systems and methods to enhance consistency of assay performance

Abstract

Systems and methods are disclosed for enhancing the consistency of performance of assays, such as multiplexed assays, by printing features in a particular pattern, such that the outer edge of the pattern has a shape that is substantially similar to the shape of the test well. For example, the pattern is a ring pattern, such that the outer edge of the ring pattern is circular or oval along the bottom of multiplexed wells. The assay substrates prepared according to the methods described result in more accurate, precise, and sensitive chemical and/or biological analyses.

1 .- 25 . (canceled) 
     
     
         26 . A testing substrate comprising:
 a substrate material defining at least one well, the at least one well having a bottom surface, a center, and at least one side wall,   the at least one side wall having a cross-sectional shape when lying in a plane parallel to the bottom surface; and   a set of analysis features on the bottom surface in a pattern, characterized in that the pattern has multiple circular rings of analysis features, wherein each ring of the multiple rings of analysis features has a shape similar to the cross-sectional shape of the side wall,   wherein for each ring of the multiple rings of analysis features, a distance from the outer edge of the at least one well to the center of each of the analysis features is about 20% to about 80% of a radius of the at least one well.   
     
     
         27 . The substrate of  claim 26 , wherein for each ring of the multiple rings of analysis features, all of the analysis features of the ring are arranged such that a distance from a center of each of the features to the center of the well is the same for each of the features. 
     
     
         28 . The substrate of  claim 26 , wherein for each ring of the multiple rings of analysis features, the analysis features of the ring are arranged such that an angle is formed by a first line that extends from the center of one feature to the center of the at least one well and a second line that extends from the center of an adjacent feature to the center of the at least one well, wherein the angle is the same for each adjacent line. 
     
     
         29 . The substrate of  claim 26 , wherein for each ring of the multiple rings of analysis features, a distance from the outer edge of the at least one well to the center of each of the analysis features is about 30% to about 70% of the radius of the at least one well. 
     
     
         30 . The substrate of  claim 26 , wherein an entirety of the bottom surface is substantially flat. 
     
     
         31 . The substrate of  claim 26 , wherein the analysis features of the set of analysis features are printed on the bottom surface. 
     
     
         32 . The substrate of  claim 26 , wherein at least one ring of the multiple rings of analysis features has 8 to 16 analysis features. 
     
     
         33 . The substrate of  claim 26 , wherein each of the analysis features of the set of analysis features has a diameter of about 250 micrometers to about 800 micrometers. 
     
     
         34 . The substrate of  claim 26 , wherein at least some of the analysis features comprise an antibody. 
     
     
         35 . A method for forming a testing substrate, comprising:
 forming a set of analysis features on the bottom surface of a well of the testing substrate, the well having a bottom surface, a center, and at least one side wall, the at least one side wall having a cross-sectional shape when lying in a plane parallel to the bottom surface, and wherein the analysis features are arranged in a pattern, characterized by multiple circular rings of analysis features, wherein each ring of the multiple rings of analysis features has a shape similar to the cross-sectional shape of the side wall,   wherein for each ring of the multiple rings of analysis features, a distance from the outer edge of the at least one well to the center of each of the analysis features is 10% to 90% of a radius of the at least one well.   
     
     
         36 . The method of  claim 35 , wherein for each ring of the multiple rings of analysis features, all of the analysis features of the ring are arranged such that a distance from a center of each of the features to the center of the well is the same for each of the features. 
     
     
         37 . The method of  claim 35 , wherein for each ring of the multiple rings of analysis features, the analysis features of the ring are arranged such that an angle is formed by a first line that extends from the center of one feature to the center of the at least one well and a second line that extends from the center of an adjacent feature to the center of the at least one well, wherein the angle is the same for each adjacent line. 
     
     
         38 . The method  claim 35 , wherein for each ring of the multiple rings of analysis features, a distance from the outer edge of the at least one well to the center of each of the analysis features is about 30% to about 70% of the radius of the at least one well. 
     
     
         39 . The method of  claim 35 , wherein an entirety of the bottom surface is substantially flat. 
     
     
         40 . The method of  claim 35 , wherein the step of forming the set of analysis features comprises printing the set of analysis features on the bottom surface. 
     
     
         41 . The method of  claim 35 , wherein at least one ring of the multiple rings of analysis features has 8 to 16 analysis features. 
     
     
         42 . The method of  claim 35 , wherein each of the analysis features of the set of analysis features has a diameter of about 250 micrometers to about 800 micrometers. 
     
     
         43 . The method of  claim 35 , wherein at least some of the analysis features comprise an antibody. 
     
     
         44 . The method of  claim 29 , further comprising:
 incubating the testing substrate;   applying blocking material to the testing substrate on which the set of analysis features is provided;   drying the testing substrate; and   processing the testing substrate for usage or storage.