US 11,719,641 B2
Spatially resolved ligand-receptor binding assays
Qiaoqiao Ruan, Kildeer, IL (US); Sylvia C. Saldana, Hinsdale, IL (US); Joseph P. Skinner, Lake Villa, IL (US); and Sergey Y. Tetin, Lindenhurst, IL (US)
Assigned to ABBOTT LABORATORIES, Abbott Park, IL (US)
Filed by Abbott Laboratories, Abbott Park, IL (US)
Filed on Apr. 1, 2022, as Appl. No. 17/711,375.
Application 17/711,375 is a continuation of application No. 16/237,098, filed on Dec. 31, 2018, granted, now 11,307,141.
Application 16/237,098 is a continuation of application No. 13/153,934, filed on Jun. 6, 2011, granted, now 10,190,986, issued on Jan. 19, 2019.
Prior Publication US 2022/0228990 A1, Jul. 21, 2022
This patent is subject to a terminal disclaimer.
Int. Cl. G01N 21/64 (2006.01); G01N 33/543 (2006.01)
CPC G01N 21/6456 (2013.01) [G01N 33/543 (2013.01)] 23 Claims
OG exemplary drawing
 
1. A method for analyzing the results of a ligand-receptor binding assay comprising the steps of:
(a) providing the results of a ligand-receptor binding assay wherein the ligand-receptor binding assay comprises combining in a reaction mixture (i) a sample suspected of containing a ligand, (ii) a first receptor attached to a microparticle that binds to the ligand, and (iii) a fluorescently labeled second receptor that binds to the ligand, and allowing formation of a complex comprising the microparticle attached to the first receptor, the ligand, and the fluorescently labeled second receptor by acquiring a white light image of the reaction mixture in order to determine the location of microparticles and a fluorescence image to determine the location of the fluorescently labeled second receptor;
(b) selecting at least one region of interest from the images wherein the at least one region of interest is a region from which light signals emanate from the complex; and
(c) qualifying the results of a ligand-receptor binding assay by:
(i) selecting pixels in the at least one region of interest for analysis;
(ii) calculating and recording the average and variance of the counts per pixel for the pixels selected in step (i);
(iii) omitting pixels that have counts greater than or less than a specified variance; and
(iv) calculating average counts per pixel of the remaining pixels.