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Isolation of a Low Number of Sperm Cells from Female DNA in a Glass–PDMS–Glass Microchip via Bead-Assisted Acoustic Differential Extraction
journal contribution
posted on 2019-01-17, 14:49 authored by Kerui Xu, Charles P. Clark, Brian L. Poe, Jenny A. Lounsbury, Johan Nilsson, Thomas Laurell, James P. LandersWe report an improved separation
method for the isolation of sperm
cells from dilute, “large volume” samples containing
female DNA using bead-assisted acoustic trapping. In an enclosed glass–PDMS–glass
(GPG) resonator, we exploit a three-layer microfluidic architecture
to generate “trapping nodes” in ultrasonic standing
waves. We investigate the dependence of trapping efficiency on particle
concentration for both sperm cells and polymeric beads. After determination
of the critical concentration of polymeric beads required to seed
the trapping event, sperm cells in dilute solution are trapped as
a result of the enhanced secondary radiation force (SRF). Sperm-cell-containing
samples with volumes up to 300 μL and cell concentrations as
low as ∼10 cells/μL are amenable to effective trapping
in the presence of an abundance of female DNA in solution. Complete
processing of samples is accomplished with separation of the female
and male fractions within 15 min. We demonstrate that the collected
fractions are amenable to subsequent DNA extraction, short tandem
repeat PCR, and the generation of STR profiles for the isolated sperm
cells.