US 12,472,495 B2
Microfluidic device for live cell manipulation and analysis
Horacio Dante Espinosa, Winnetka, IL (US); Milan Mrksich, Hinsdale, IL (US); Prithvijit Mukherjee, Evanston, IL (US); Eric Jason Berns, Naperville, IL (US); and Cesar Andres Patino, Evanston, IL (US)
Assigned to Northwestern University, Evanston, IL (US)
Filed by NORTHWESTERN UNIVERSITY, Evanston, IL (US)
Filed on Oct. 26, 2021, as Appl. No. 17/511,246.
Claims priority of provisional application 63/106,680, filed on Oct. 28, 2020.
Prior Publication US 2022/0126292 A1, Apr. 28, 2022
Int. Cl. B01L 3/00 (2006.01)
CPC B01L 3/502715 (2013.01) [B01L 3/502707 (2013.01); B01L 3/502753 (2013.01); B01L 2200/12 (2013.01); B01L 2300/0663 (2013.01); B01L 2300/0829 (2013.01); B01L 2300/0887 (2013.01); B01L 2300/123 (2013.01); B01L 2300/16 (2013.01)] 11 Claims
OG exemplary drawing
 
1. A cell analysis system comprising:
a multi-layer microfluidic device comprising:
a first side and a second side;
a microfluidic channel;
a first layer including a plurality of microwells disposed underneath the microfluidic channel and in fluidic communication with the microfluidic channel, wherein each microwell of the plurality of microwells is configured to culture one or more cells;
an intermediate conductive layer disposed on top of the plurality of microwells;
a second layer including a plurality of extraction chambers disposed underneath the first layer wherein each microwell of the plurality of microwells is substantially concentric with a corresponding extraction chamber of the plurality of extraction chambers; and
a membrane disposed between the first layer and the second layer, wherein the membrane has a plurality of nanochannel pores enabling the plurality of microwells to fluidically communicate with the plurality of extraction chambers;
a top conductive layer disposed on the first side of the multi-layer microfluidic device and a bottom conductive layer disposed on the second side of the multi-layer microfluidic device wherein the second layer is opposite to the first side such that the multi-layer microfluidic device is sandwiched between the top conductive layer of the cell analysis system and the bottom conductive layer of the cell analysis system;
one or more sensors disposed on the bottom conductive layer, projecting into each extraction chamber of the plurality of extraction chambers and for analyzing intracellular contents in the plurality of extraction chambers originating from the plurality of microwells and diffusing into the plurality of extraction chambers through the plurality of nanochannel pores; and
wherein the cell analysis system further comprises:
a function generator configured to apply an electroporation pulse to the one or more cells within the plurality of microwells between the top conductive layer and the bottom conductive layer, wherein the intermediate conductive layer disposed on top of the plurality of microwells is configured to enhance electrical conductivity and minimize electric field losses in the microfluidic channel when the electroporation pulse is applied.