US 12,447,469 B2
Methods and apparatus for single biological nanoparticle analysis
Daniel T. Chiu, Seattle, WA (US); Seung Ryoung Jung, Shoreline, WA (US); Jingang Wang, Bothell, WA (US); and Bryant S. Fujimoto, Seattle, WA (US)
Assigned to University of Washington, Seattle, WA (US)
Appl. No. 15/733,720
Filed by UNIVERSITY OF WASHINGTON, Seattle, WA (US)
PCT Filed Mar. 29, 2019, PCT No. PCT/US2019/025033
§ 371(c)(1), (2) Date Oct. 2, 2020,
PCT Pub. No. WO2019/199499, PCT Pub. Date Oct. 17, 2019.
Claims priority of provisional application 62/657,278, filed on Apr. 13, 2018.
Prior Publication US 2021/0016276 A1, Jan. 21, 2021
Int. Cl. B01L 3/00 (2006.01); G01N 15/14 (2024.01); G01N 33/543 (2006.01); B82Y 35/00 (2011.01); G01N 15/10 (2006.01)
CPC B01L 3/502715 (2013.01) [B01L 3/502776 (2013.01); G01N 15/1459 (2013.01); G01N 33/54366 (2013.01); B01L 2200/0636 (2013.01); B01L 2200/0647 (2013.01); B82Y 35/00 (2013.01); G01N 2015/1006 (2013.01); G01N 2015/1493 (2013.01)] 18 Claims
OG exemplary drawing
 
1. A method for determining the size of a biological nanoparticle in a fluid sample, the method comprising:
providing a planar microfluidic chip comprising at least one microfluidic channel, wherein the at least one microfluidic channel comprises at least one constriction comprising a detection region;
introducing the fluid sample into the planar microfluidic chip, the fluid sample comprising a plurality of biological nanoparticles;
flowing a portion of the plurality of biological nanoparticles through the at least one microfluidic channel;
illuminating in the detection region of the at least one microfluidic channel at least one biological nanoparticle from the portion of the plurality of biological nanoparticles on a particle-by-particle basis, wherein the illuminating of the at least one biological nanoparticle comprises using an illumination source having a beam width of less than 2 μm within the at least one microfluidic channel, and wherein the illumination source illuminates the detection region within the at least one microfluidic channel comprising a cross sectional area of less than 10 μm2;
detecting epifluorescent light intensity emitted from the at least one biological nanoparticle; and
assigning a size value to the at least one illuminated biological nanoparticle based on the detected epifluorescent light intensity, wherein a biological nanoparticle with a larger hydrodynamic radius emits a higher intensity of light compared to a biological nanoparticle with a smaller hydrodynamic radius, and wherein the biological nanoparticle has a hydrodynamic diameter of less than 1 μm.