US 12,000,842 B2
Systems and methods for generating droplets and performing digital analyses
Hei Mun Christina Fan, Palo Alto, CA (US); Janice Hoiyi Lai, Mountain View, CA (US); Sixing Li, Mountain View, CA (US); Stephen P. A. Fodor, Palo Alto, CA (US); and Eleen Yee Lam Shum, San Carlos, CA (US)
Assigned to ENUMERIX, INC., Palo Alto, CA (US)
Filed by Enumerix, Inc., Palo Alto, CA (US)
Filed on Apr. 1, 2022, as Appl. No. 17/711,417.
Application 17/711,417 is a continuation of application No. PCT/US2022/018994, filed on Mar. 4, 2022.
Claims priority of provisional application 63/157,292, filed on Mar. 5, 2021.
Prior Publication US 2022/0283174 A1, Sep. 8, 2022
This patent is subject to a terminal disclaimer.
Int. Cl. G01N 33/68 (2006.01); C12Q 1/6844 (2018.01); G01N 21/64 (2006.01)
CPC G01N 33/689 (2013.01) [C12Q 1/6844 (2013.01); G01N 21/645 (2013.01); G01N 21/6486 (2013.01); G01N 2021/6482 (2013.01)] 16 Claims
OG exemplary drawing
 
1. A method comprising:
performing a digital analysis of target nucleic acid material from a sample, wherein performing the digital analysis comprises:
(a) generating an emulsion within a collection container, wherein the emulsion comprises a plurality of droplets, wherein each droplet of the plurality of droplets is surrounded by an immiscible film, and wherein the plurality of droplets is in a continuous phase immiscible with the immiscible film, and wherein the plurality of droplets comprises at least 20 million droplets, wherein the at least 20 million droplets comprise portions of the sample and amplification reaction reagents, and wherein a droplet of the at least 20 million droplets comprises the target nucleic acid material;
(b) amplifying the target nucleic acid material in the droplet; and
(c) optically interrogating via light sheet imaging the emulsion within the collection container comprising the at least 20 million droplets across a plurality of cross-sections of the emulsion, wherein (a)-(c) are performed in less than or equal to 2.5 hours.