US 12,281,355 B2
Multiplexed imaging with enzyme mediated amplification
Grady Carlson, Houston, TX (US)
Assigned to Akoya Biosciences, Inc., Menlo Park, CA (US)
Filed by Akoya Biosciences, Inc., Menlo Park, CA (US)
Filed on Sep. 30, 2020, as Appl. No. 17/039,888.
Claims priority of provisional application 62/908,540, filed on Sep. 30, 2019.
Prior Publication US 2021/0222234 A1, Jul. 22, 2021
Int. Cl. C12Q 1/6832 (2018.01); C12Q 1/6804 (2018.01); G01N 1/30 (2006.01)
CPC C12Q 1/6832 (2013.01) [C12Q 1/6804 (2013.01); G01N 1/30 (2013.01)] 32 Claims
 
1. A method for imaging an analyte in a biological sample, the method comprising:
contacting the biological sample with a binding agent, wherein the binding agent comprises a binding moiety that binds to the analyte and a first oligonucleotide sequence;
contacting the biological sample with a catalytic agent, wherein the catalytic agent comprises a second oligonucleotide sequence linked to an enzyme, and wherein the second oligonucleotide sequence hybridizes to the first oligonucleotide sequence;
contacting the biological sample with a localization agent, wherein the localization agent comprises a substrate complementary to the enzyme and a third oligonucleotide sequence linked to the substrate;
contacting the biological sample with a labeling agent, wherein the labeling agent comprises a fourth oligonucleotide sequence linked to an optical label, wherein the fourth oligonucleotide sequence hybridizes to the third oligonucleotide sequence; and
exposing the biological sample to illumination light, detecting emitted light from the biological sample, and forming an image of the biological sample in which a location of the analyte is indicated by the optical label.