US 12,404,551 B2
Single molecule analysis in an electrical field
Rudolf Rigler, Djursholm (SE)
Assigned to GNOTHIS HOLDING AG, Cham (CH)
Filed by GNOTHIS HOLDING AG, Cham (CH)
Filed on Nov. 2, 2022, as Appl. No. 18/052,004.
Application 18/052,004 is a division of application No. 15/740,903, granted, now 11,519,030, previously published as PCT/EP2016/065019, filed on Jun. 28, 2016.
Claims priority of application No. 15174524 (EP), filed on Jun. 30, 2015.
Prior Publication US 2023/0304087 A1, Sep. 28, 2023
Int. Cl. C12Q 1/6874 (2018.01); B01L 3/00 (2006.01); G01N 1/40 (2006.01); G01N 21/64 (2006.01); G01N 15/10 (2006.01)
CPC C12Q 1/6874 (2013.01) [B01L 3/5085 (2013.01); G01N 1/40 (2013.01); G01N 21/6428 (2013.01); G01N 21/648 (2013.01); B01L 2300/0645 (2013.01); B01L 2300/0819 (2013.01); G01N 2001/4038 (2013.01); G01N 2015/1006 (2013.01); G01N 2015/1028 (2024.01); G01N 21/6408 (2013.01); G01N 2021/6439 (2013.01)] 16 Claims
 
1. A device for analyzing at least one individual single molecule, and/or for sequencing at least one single nucleic acid molecule, the device comprising:
(i) a planar support comprising at least one individual sample nano-spot thereon, wherein said at least one individual sample nano-spot is at least partially made from an electrically conductive material,
(ii) means adapted for contacting said support with a liquid medium comprising at least one single molecule to be analysed,
(iii) means adapted for applying an electrical field across a reaction space which is formed upon contacting the support with liquid medium, wherein the electrical field is applied with a direct current voltage, whereby a concentration of said at least one single molecule to be analyzed at said sample nano-spot is effected,
(iv) a light source which provides at least one individual illuminated volume element at said sample nano-spot on the support, for individually illuminating the single molecule at said individual sample nano-spot,
(v) a light detector for individually detecting light emitted from the single molecule at an individual sample nano-spot,
(vi) means for carrying out an optical projection of a detection pixel onto the support and aligning an individual sample nano spot to the projection of a single detection pixel on the support, wherein the optical projection of the detection pixel on the support has a diameter in the range of about 100 nm to 5 μm, and
(vii) means for adjusting the alignment between the sample nano spot and the detection pixel projection,
wherein the at least one individual illuminated volume element is an evanescent field generated by total internal reflection.