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    Area of Science:

    • Biotechnology
    • Microfluidics
    • Bioanalytical Chemistry

    Background:

    • Biological samples often have complex topography, hindering precise reagent delivery.
    • Existing methods can be intrusive or lack depth control for microscale alterations.

    Purpose of the Study:

    • To introduce a nonintrusive method for localized reagent delivery on immersed biological samples.
    • To demonstrate deep-reaching hydrodynamic flow confinement (DR-HFC) for precise microscale alterations on varied substrates.

    Main Methods:

    • Developed a passive microfluidic probe design for DR-HFC, featuring reagent-injection and aspiration apertures.
    • Utilized a large gap between the probe and substrate to prevent sample intrusion and reduce shear stress.
    • Investigated linear and annular DR-HFC probe designs and their parameters numerically.

    Main Results:

    • Successfully demonstrated localized binding of antihuman immunoglobulin G (IgG) at depths from 50 to 600 μm.
    • Validated the DR-HFC technique's ability to control reagent localization and depth on topographical substrates.
    • Showcased the method's capability to scan centimeter-scale areas passively.

    Conclusions:

    • DR-HFC offers a simple, nonintrusive approach for precise reagent delivery on biological samples with topographical variations.
    • The technique is readily implementable and suitable for next-generation diagnostic and bioanalytical devices.
    • DR-HFC minimizes sample damage by maintaining a gap and reducing shear stress.