US 12,435,361 B2
Simultaneous quantification of a plurality of proteins in a user-defined region of a cross-sectioned tissue
Joseph M. Beechem, Eugene, OR (US); Gordon B. Mills, Houston, TX (US); Charles Warren, Bremerton, WA (US); Chris Merritt, Seattle, WA (US); Jaemyeong Jung, Bellevue, WA (US); Dwayne L. Dunaway, Seattle, WA (US); and Scott Crowder, Seattle, WA (US)
Assigned to Bruker Spatial Biology, Inc., Seattle, WA (US); and Board of Regents, The University of Texas System, Austin, TX (US)
Filed by Bruker Spatial Biology, Inc., Seattle, WA (US); and Board of Regents, The University of Texas System, Austin, TX (US)
Filed on Oct. 8, 2019, as Appl. No. 16/596,587.
Application 16/596,587 is a continuation of application No. 15/211,236, filed on Jul. 15, 2016, granted, now 10,501,777.
Claims priority of provisional application 62/323,018, filed on Apr. 15, 2016.
Claims priority of provisional application 62/277,283, filed on Jan. 11, 2016.
Claims priority of provisional application 62/261,654, filed on Dec. 1, 2015.
Claims priority of provisional application 62/193,819, filed on Jul. 17, 2015.
Prior Publication US 2020/0040382 A1, Feb. 6, 2020
Int. Cl. C12Q 1/6804 (2018.01); C12Q 1/6832 (2018.01); C12Q 1/6841 (2018.01); G01N 33/543 (2006.01); G01N 33/58 (2006.01)
CPC C12Q 1/6832 (2013.01) [C12Q 1/6804 (2013.01); C12Q 1/6841 (2013.01); G01N 33/54306 (2013.01); G01N 33/58 (2013.01); G01N 2458/10 (2013.01)] 20 Claims
OG exemplary drawing
 
1. A method comprising:
(1) contacting at least one protein target in a tissue sample with at least one probe comprising a target-binding domain, a signal oligonucleotide and a photo-cleavable motif located between the target binding domain and the signal oligonucleotide, wherein the tissue sample is immersed in fluid;
(2) illuminating a specific location of the immersed tissue sample with light sufficient to release the signal oligonucleotide into the fluid;
(3) collecting the released signal oligonucleotide by aspirating the fluid proximal to the specific location that was illuminated, wherein the fluid proximal to the specific location that was illuminated is located directly above the specific location that was illuminated, wherein the aspiration is performed using a capillary tube no more than 100 μm above the immersed tissue sample; and
(4) identifying the released signal oligonucleotide, thereby detecting the at least one protein target in the specific location of the tissue sample that was illuminated.