US 11,948,302 B2
Automated holographic video microscopy assay
David G. Grier, New York, NY (US); Fook Chiong Cheong, Brooklyn, NY (US); Kaitlynn Snyder, New York, NY (US); Rushna Quddus, New York, NY (US); Lauren E. Altman, New York, NY (US); and Kent Kirshenbaum, New York, NY (US)
Assigned to New York University, New York, NY (US); and Spheryx, Inc, New York, NY (US)
Filed by NEW YORK UNIVERSITY, New York, NY (US); and SPHERYX, INC., New York, NY (US)
Filed on Mar. 9, 2021, as Appl. No. 17/196,895.
Claims priority of provisional application 62/987,223, filed on Mar. 9, 2020.
Prior Publication US 2021/0279876 A1, Sep. 9, 2021
Int. Cl. G06T 7/00 (2017.01); G01N 15/1434 (2024.01); G02B 21/36 (2006.01); G03H 1/00 (2006.01); G03H 1/04 (2006.01); G06T 7/62 (2017.01)
CPC G06T 7/0014 (2013.01) [G01N 15/1434 (2013.01); G02B 21/365 (2013.01); G03H 1/0005 (2013.01); G03H 1/0465 (2013.01); G06T 7/62 (2017.01); G01N 2015/1445 (2013.01); G03H 2001/005 (2013.01); G03H 2001/0471 (2013.01); G03H 2210/55 (2013.01); G03H 2222/13 (2013.01); G03H 2222/34 (2013.01); G06T 2207/10056 (2013.01); G06T 2207/30024 (2013.01)] 20 Claims
OG exemplary drawing
 
1. A method of characterizing a parameter of a viral protein by holographic microscopy, comprising the steps of:
receiving holographic image data having a set of concentric bright and dark rings associated with a functionalized probe bead;
determining by Lorenz-Mie analysis an effective sphere estimate of each of the functionalized probe bead's effective sphere diameter, and effective sphere refractive index; and
determining a diameter difference between the effective sphere diameter with a predetermined diameter for unbound functionalized probe beads, with diameter difference representing the result of material bound to the bead and medium within the effective sphere estimate, wherein the effective sphere diameter (dp) is equal to
dp=d0+2δf
wherein dp is the effective sphere diameter, d0 is the predetermined diameter for unbound functionalized probe beads, δ is effective optical thickness of a complete layer of the viral protein, and f is the fraction of binding sites on the functionalized probe bead.