	US 12,454,716 B2
	Bioluminescent single photon bioreactor and performing absolute quantification of light-producing activity by enzymes
Sergey Vladimirovich Polyakov, Gaithersburg, MD (US); and Ralph Jimenez, Boulder, CO (US)
Assigned to Government of the United States of America, as represented by Secretary of Commerce, Gaithersburg, MD (US)
Filed by Government of the United States of America, as represented by the Secretary of Commerce, Gaithersburg, MD (US)
Filed on Nov. 1, 2021, as Appl. No. 17/515,832.
Claims priority of provisional application 63/186,372, filed on May 10, 2021.
Prior Publication US 2022/0364144 A1, Nov. 17, 2022
Int. Cl. C12Q 1/66 (2006.01); B01L 3/00 (2006.01); C12N 11/14 (2006.01); G01N 21/76 (2006.01)

CPC C12Q 1/66 (2013.01) [B01L 3/502 (2013.01); B01L 3/508 (2013.01); C12N 11/14 (2013.01); C12Y 113/12013 (2013.01); G01N 21/763 (2013.01); B01L 2200/026 (2013.01); B01L 2300/023 (2013.01); B01L 2300/0654 (2013.01); B01L 2300/0663 (2013.01)]

12 Claims

1. A method implemented by a system of one or more processors, the system being included in a bioluminescent single photon bioreactor and for performing absolute quantification of light-producing activity by enzymes by the bioluminescent single photon bioreactor, the method comprising:

producing, by a pixel detector of the bioluminescent single photon bioreactor, a detector signal;

receiving, by an analyzer of the bioluminescent single photon bioreactor, the detector signal;

determining, by the bioluminescent single photon bioreactor, a time of arrival of the detector signal;

determining, by the bioluminescent single photon bioreactor, an autocorrelation among detector signals for threshold times of arrival of photons at pixel detector according to:

the following equation for a single emitter in the field of view of an objective of the bioluminescent single photon bioreactor:

g⁽²⁾(0) for the number of emitters N in the field of view (FOV) of the objective, such that:

if g⁽²⁾(0) is 0, then one enzyme was in the field of view; and

if g⁽²⁾(0) is not 0, then the number of emitters N is greater than 1, or noise is present in the detector signal.