	US 7,499,806 B2
	Image processing in microsphere arrays
Bahram Ghaffarzadeh Kermani, San Diego, Calif. (US); and Juergen Haas, San Diego, Calif. (US)
Assigned to Illumina, Inc., San Diego, Calif. (US)
Appl. No. 10/504,568 PCT Filed Feb. 14, 2003, PCT No. PCT/US03/04570 § 371(c)(1), (2), (4) Date May 06, 2005, PCT Pub. No. WO03/069333, PCT Pub. Date Aug. 21, 2003.
Claims priority of provisional application 60/357213, filed on Feb. 14, 2002.
Prior Publication US 2006/0041384 A1, Feb. 23, 2006
Int. Cl. G06F 17/11 (2006.01); G06F 17/50 (2006.01); C12Q 1/68 (2006.01); C12M 1/00 (2006.01)

U.S. Cl. 702—19 [702/28; 702/32; 382/132; 435/6; 435/71; 435/174; 435/283.1; 435/287.2]

11 Claims

1. A method of registering an analytical image of a microsphere array comprising:

a) providing a microsphere array, wherein said microsphere array comprises:

i) a substrate with a surface comprising discrete sites; and

ii) a population of microspheres comprising at least a first and a second subpopulation each comprising a bioactive agent, wherein said population of microspheres is distributed on said surface such that each of said discrete sites contain no more than 1 microsphere;

b) decoding said microsphere array so that the location of each bioactive agent is elucidated;

c) computing a registration grid based on known locations of said bioactive agents from information obtained from step (b);

d) contacting said array with a sample comprising target analytes;

e) acquiring a hybridization intensity image of said microsphere array, wherein said hybridization intensity image comprises a first bright bead type to serve as a fiducial;

f) overlaying the registration grid onto said hybridization intensity image;

g) determining an average intensity for the first bright bead type; and

h) positioning the registration grid such that the average intensity of the first bright bead type is maximal, thereby registering the analytical image of the microsphere array.