Abstract: A method for producing an image of an object within a chamber of a microfluidic device includes providing the microfluidic device having x, y, and z dimensions and a chamber depth center point located along the z dimension. The chamber depth center point is located a known z dimension distance from a fiducial marking embedded within the microfluidic device. The method also includes placing the microfluidic device within an imaging system that includes an optical device capable of detecting the fiducial marking. The optical device defines an optical path axially aligned with the z dimension and has a focal plane perpendicular to the optical path. When the focal plane is moved along the optical path, the fiducial marking is maximally detected when the focal plane is at the z depth in comparison to when the focal plane is not substantially in-plane with the z depth.

Abstract: A biological substrate, e.g., microfluidic chip. The substrate includes a rigid substrate material, which has a surface region capable of acting as a handle substrate. The substrate also has a deformable fluid layer coupled to the surface region. One or more well regions are formed in a first portion of the deformable fluid layer and are capable of holding a fluid therein. The one or more channel regions are formed in a second portion of the deformable fluid layer and are coupled to one or more of the well regions. An active region is formed in the deformable fluid layer. At least three fiducial markings are formed within the non-active region and disposed in a spatial manner associated with at least one of the well regions. A control layer is coupled to the fluid layer.