Deep-reaching hydrodynamic flow confinement: Micrometer-scale liquid localization for open substrates with topographical variations
We present a method for nonintrusive localization and reagent delivery on immersed biological samples with topographical variation on the order of hundreds of micrometers. Our technique, which we refer to as the deepreaching hydrodynamic flow confinement (DR-HFC), is simple and passive: it relies on a deep-reaching hydrodynamic confinement delivered through a simple microfluidic probe design to perform localized microscale alterations on substrates as deep as 600 μm. Designed to scan centimeterscale areas of biological substrates, our method passively prevents sample intrusion by maintaining a large gap between the probe and the substrate. The gap prevents collision of the probe and the substrate and reduces the shear stress experienced by the sample. We present two probe designs: linear and annular DR-HFC. Both designs comprise a reagent-injection aperture and aspiration apertures that serve to confine the reagent. We identify the design parameters affecting reagent localization and depth by DRHFC and study their individual influence on the operation of DR-HFC numerically. Using DR-HFC, we demonstrate localized binding of antihuman immunoglobulin G (IgG) onto an activated substrate at various depths from 50 to 600 μm. DR-HFC provides a readily implementable approach for noninvasive processing of biological samples applicable to the next generation of diagnostic and bioanalytical devices.