US 12,305,232 B2
Single molecule analysis with high accuracy
Rudolf Rigler, Djursholm (SE); and Lars Edman, Stockholm (SE)
Assigned to GNOTHIS HOLDING AG, Cham (CH)
Filed by GNOTHIS HOLDING AG, Zug (CH)
Filed on May 22, 2023, as Appl. No. 18/200,109.
Application 18/200,109 is a division of application No. 17/376,324, filed on Jul. 15, 2021, granted, now 11,788,140.
Application 17/376,324 is a division of application No. 15/110,779, granted, now 11,091,805, issued on Aug. 17, 2021, previously published as PCT/EP2015/050066, filed on Jan. 5, 2015.
Claims priority of application No. 14150807 (EP), filed on Jan. 10, 2014.
Prior Publication US 2023/0295720 A1, Sep. 21, 2023
Int. Cl. C12Q 1/6874 (2018.01); C12Q 1/6837 (2018.01); G01N 21/64 (2006.01)
CPC C12Q 1/6874 (2013.01) [C12Q 1/6837 (2013.01); G01N 21/6428 (2013.01); G01N 21/6452 (2013.01); G01N 2021/6439 (2013.01)] 23 Claims
 
1. A device for analyzing at least one individual single molecule, comprising:
(a) a support for positioning a single molecule to be analyzed on an individual spot, comprising at least one individual sample spot, wherein said spot has a diameter in the range of about 1 nm-20 nm and the distance between individual spots is at least about 5 times the diameter of the spot,
(b) a light source which provides at least one individual illuminated volume element at an individual sample spot on the support, for illuminating a single molecule at an individual spot,
(c) a light detector which comprises at least one detection pixel wherein said detection pixel has a diameter in the range of about 0.5 μm-50 μm and the distance between said detection pixels is at least about 2 times the diameter of the detection pixel, for individually detecting light emitted from a single molecule at an individual spot,
(d) means for optically projecting said detection pixel onto the support, comprising an optical pathway between the light source, the support and the light detector, wherein the optical projection has a diameter in the range of about 100 nm-5 μm and is about 10-200 times smaller than the size of the detection pixel on the detector,
(e) means for aligning an individual sample spot on the support to the optical projection of a single detection pixel such that the center of the individual sample spot is aligned to the projection of the single detection pixel on the support, wherein the distance between individual sample spots on the support is equivalent to the distance between optical projections of detection pixels on the support, and
(f) means for detecting the signals from individual detection pixels, wherein said signals result from an event associated with a single molecule positioned on an individual spot.