Microfluidic dual-well device for highthroughput single-cell capture and culture

A microfluidic dual-well device comprising:

• (a) a first substrate, having a first end, a second end opposite to the first end, and a culture microwell forming portion with a thickness of t1 located between the two ends, each end having a first surface and a second surface and a thickness of T1, the culture microwell forming portion having a first surface and a second surface opposite to the first surface, wherein the thickness T1 of each end of the first substrate is greater than the thickness t1 of the culture microwell forming portion;
• (b) a plurality of culture microwells, each microwell having a diameter of w1 and a depth of d1, extending from the second surface toward the first surface of the culture microwell forming portion;
• (c) a second substrate with a thickness of t2, having a first surface and a second surface opposite to the first surface of the second substrate, the second substrate having a first end, a second end opposite to the first end, and a capture microwell forming portion located between the two ends of the second substrate, the two ends of the second substrate being respectively bounded to the two ends of the first substrate with its second surface facing toward the second surface of the first substrate at a distance of h;
• (d) a plurality of capture microwells, each microwell having a diameter of w2 and a depth of d2, extending from the second surface toward the first surface of the second substrate, wherein the capture microwells are in alignment with the culture microwells so that each microwell is corresponding to one culture microwell and the projected area of each capture microwell is within the projected area of the corresponding culture well, wherein the diameter w2 of each capture microwell is smaller than the diameter w1 of each culture microwells;
• (e) a microfluidic channel with a length of l, a width of w3 and a height of h, formed between the two second surfaces of culture and capture microwell forming portions;
• (f) a microfluidic inlet port defining an opening in the first surface of the first substrate;
• and
• (g) a microfluidic outlet port defining an opening in the first surface of the first substrate, opposite to the microfluidic inlet port;
• wherein the microfluidic channel is in fluidic connections with the culture microwells, the capture microwells, and the inlet and outlet ports.