Apparatuses and methods for pathogen detection using microfluidic biochips

A hybrid microfluidic microplate apparatus comprising:

• a top layer comprising at least one inlet reservoir coupled to a microchannel,
• a middle layer positioned beneath the top layer, said middle layer comprises at least one funnel shaped well comprising an upper well portion having a first diameter of 1 mm (millimeter) to 5 mm and a lower well portion having a second diameter of 0.1 mm to 1 mm, wherein a top portion of the upper well portion is connected to the microchannel in the top layer, and a bottom portion of the upper well portion is connected to a top portion of the lower well portion through a junction, and the second diameter is smaller than the first diameter,
• an insert positioned inside the upper well portion and over the junction of the upper well portion and the lower well portion, said insert forms a separate layer within the at least one funnel shaped well and is made of a first material comprising a porous paper comprising cellulose, nitrocellulose, nylon, or any combinations thereof, and
• a bottom layer positioned beneath the middle layer, said bottom layer comprises an outlet hole coupled to at least one outlet channel, said at least one outlet channel is coupled to a bottom portion of the lower well portion, and the outlet hole is fluidly connected to the inlet reservoir via the microchannel in the top layer, upper well portion, lower well portion, and at least one outlet channel,
• wherein the top layer, the middle layer and the bottom layer is made with a second material comprising a polymer.