Abstract: A multi-compartment microfluidic device for enabling fluidic isolation among interconnected compartments and accomplishing centrifugal positioning and/or patterned substrate positioning of biological specimens. Includes micropatterned substrate coupled with optically transparent housing allowing imaging. Housing includes microfluidic region having entry reservoir for accepting first volume of fluid and additional microfluidic region(s) having a second entry reservoir for accepting second volume of fluid less than first volume of fluid to create hydrostatic pressure. A barrier region that couples the microfluidic region with the second microfluidic region enables biological specimen(s) to extend across the microfluidic, barrier region and second microfluidic region. The barrier region includes embedded microgroove(s) having width and height enabling second volume of fluid to be fluidically isolated from first volume of fluid via hydrostatic pressure maintained via the embedded microgroove(s).