US 11,988,604 B2
Optical microscopy with phototransformable optical labels
Robert Eric Betzig, Berkeley, CA (US); and Harald F. Hess, Leesburg, VA (US)
Assigned to Hestzig LLC, Ashburn, VA (US)
Filed by Hestzig LLC, Leesburg, VA (US)
Filed on May 13, 2021, as Appl. No. 17/302,829.
Application 17/302,829 is a continuation of application No. 16/166,485, filed on Oct. 22, 2018, granted, now 11,009,460.
Application 16/166,485 is a continuation of application No. 15/170,521, filed on Jun. 1, 2016, granted, now 10,107,753, issued on Oct. 23, 2018.
Application 15/170,521 is a continuation of application No. 14/064,582, filed on Oct. 28, 2013, granted, now 9,360,426, issued on Jun. 7, 2016.
Application 14/064,582 is a continuation of application No. 13/655,102, filed on Oct. 18, 2012, granted, now 8,599,376, issued on Dec. 3, 2013.
Application 13/655,102 is a continuation of application No. 12/956,946, filed on Nov. 30, 2010, granted, now 8,462,336, issued on Jun. 11, 2013.
Application 12/956,946 is a continuation of application No. 12/645,019, filed on Dec. 22, 2009, granted, now 7,864,314, issued on Jan. 4, 2011.
Application 12/645,019 is a continuation of application No. 11/944,274, filed on Nov. 21, 2007, granted, now 7,782,457, issued on Aug. 24, 2010.
Application 11/944,274 is a continuation of application No. PCT/US2006/019887, filed on May 23, 2006.
Claims priority of provisional application 60/780,968, filed on Mar. 10, 2006.
Claims priority of provisional application 60/683,337, filed on May 23, 2005.
Prior Publication US 2022/0091038 A1, Mar. 24, 2022
This patent is subject to a terminal disclaimer.
Int. Cl. G01N 21/64 (2006.01); G01N 33/58 (2006.01); G02B 21/16 (2006.01); G02B 21/36 (2006.01); G02B 27/58 (2006.01)
CPC G01N 21/6428 (2013.01) [G01N 21/64 (2013.01); G01N 21/6458 (2013.01); G01N 33/582 (2013.01); G02B 21/16 (2013.01); G02B 21/367 (2013.01); G02B 27/58 (2013.01); G01N 2021/6419 (2013.01); G01N 2021/6421 (2013.01); G01N 2021/6439 (2013.01); G01N 2021/6441 (2013.01); G01N 21/648 (2013.01)] 25 Claims
OG exemplary drawing
 
1. A method comprising:
(a) providing activation radiation to a sample that includes phototransformable optical labels (“PTOLs”) to activate a subset of the PTOLs in the sample, wherein the activation radiation is provided such that adjacent activated PTOLs in a focal plane of a detection objective of imaging optics used to image radiation from the PTOLs are separated by distances that are, on average, larger than a diffraction limited resolution of the detection objective;
(b) providing excitation radiation to the sample to excite at least some of the activated subset of PTOLs;
(c) detecting radiation emitted from an activated and excited PTOL of the subset of PTOLs, wherein the activated and excited PTOL is bound to a protein in the sample;
(d) repeating (b) and (c) at different times; and
(e) determining, based on the detected radiation, locations of the activated and excited PTOL bound to the protein at the different times, wherein the locations are determined with an accuracy that is greater than a diffraction limited resolution of the detection objective.