US 12,455,224 B2
Single particle automated Raman trapping analysis
Molly Stevens, London (GB); Jelle Penders, London (GB); and Isaac Pence, London (GB)
Assigned to Sparta Biodiscovery Limited, London (GB)
Appl. No. 17/252,891
Filed by Imperial College Innovations Limited, London (GB)
PCT Filed Jun. 18, 2019, PCT No. PCT/EP2019/066106
§ 371(c)(1), (2) Date Dec. 16, 2020,
PCT Pub. No. WO2019/243375, PCT Pub. Date Dec. 26, 2019.
Claims priority of application No. 1810010 (GB), filed on Jun. 19, 2018.
Prior Publication US 2021/0262915 A1, Aug. 26, 2021
Int. Cl. G01N 15/1434 (2024.01); G01N 15/10 (2024.01); G01N 15/14 (2024.01); G01N 21/65 (2006.01)
CPC G01N 15/1434 (2013.01) [G01N 15/1012 (2013.01); G01N 21/65 (2013.01); G01N 2015/1493 (2013.01)] 28 Claims
OG exemplary drawing
 
1. A method of automated particle analysis using an apparatus for particle trapping and data acquisition, comprising:
(i) performing, using a control system, a calibration procedure for determining a threshold indicative of a particle capture of a target particle from a particle-conveying medium, the calibration procedure comprising establishing, using the control system, a spectral feature which differentiates between (i) a Raman response of the target particle to an electromagnetic radiation beam generated using a single electromagnetic radiation source and (ii) a Raman response of the particle-conveying medium without the target particle to the electromagnetic radiation beam, in which the calibration procedure comprises determining a spectral profile using at least a part of a Raman response spectrum that provides a differentiation, wherein the apparatus comprises the single electromagnetic radiation source, a focusing element, a detector, and the control system configured to control at least the single electromagnetic radiation source and the detector;
(ii) focusing, using the focusing element, the electromagnetic radiation beam onto the particle-conveying medium, the electromagnetic radiation beam having a waist portion at its focus which defines a particle capture zone for capturing a particle within the electromagnetic radiation beam;
(iii) automatically performing, by the processor using the detector, a first data acquisition procedure to test for the particle capture of a target particle from the particle-conveying medium, in which the first data acquisition procedure comprises performing a Raman response data gathering procedure sufficient to detect a presence of the spectral profile determined from the calibration procedure above the threshold indicative of the particle capture of a target particle from the particle-conveying medium;
(iv) if the particle capture of a target particle from the particle-conveying medium is not detected, automatically repeating step (iii);
(v) if the particle capture of a target particle from the particle-conveying medium is detected, automatically acquiring, using the detector, particle data from the captured particle;
(vi) automatically reducing, using the electromagnetic radiation source or a modulation device, an intensity of the electromagnetic radiation beam to a sub-capture level to release the particle from the particle capture zone; and
(vii) automatically repeating steps (ii) to (vi) for successive particles in the particle-conveying medium.