MULTIPLEX ANALYSIS OF MOLECULES IN SINGLE CELLS BY IMAGE CORRELATION

摘要

Disclosed herein are methods and systems for analyzing visual data from multiple rounds of hybridization interactions where the same molecular target is detected by probes with different detectable labels. In particular, disclosed herein are methods and systems for analyzing sequential hybridization images for molecular profiling, where the images are obtained using multiplex fluorescence in situ hybridization (FISH).

主权项

1. A method of determining an abundance level of a molecular target within a defined region in a cell, comprising:
identifying, in the defined region in the cell, a copy of the molecular target by spatially correlating visual data from a plurality of hybridizations {1, 2, . . . N}, wherein N is equal or greater than 2 and wherein the plurality of hybridizations comprises:
a first round of hybridization during which a first plurality of probes is applied to the cell, wherein the first plurality comprises one or more first probes interacting with the molecular target to provide a first visual signal at least partially constituting the visual data for the first round of hybridization; anda second round of hybridization during which a second plurality of probes is applied to the cell, wherein the second plurality comprises one or more second probes interacting with the same copy of the molecular target to provide a second visual signal constituting the visual data for the second round of hybridization, and wherein the second visual signal differs from the first visual signal; and determining an abundance level of the molecular target based on a correlation function between the visual data for the first round of hybridization and the visual data for the second round of hybridization.