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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

Xu, Kerui ; Clark, Charles P. ; Poe, Brian L. ; Lounsbury, Jenny A. ; Nilsson, Johan LU ; Laurell, Thomas LU and Landers, James P. (2019) In Analytical Chemistry 91(3). p.2186-2191
Abstract

We 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... (More)

We 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.

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organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Analytical Chemistry
volume
91
issue
3
pages
2186 - 2191
publisher
The American Chemical Society (ACS)
external identifiers
  • pmid:30652864
  • scopus:85060197044
ISSN
0003-2700
DOI
10.1021/acs.analchem.8b04752
language
English
LU publication?
yes
id
7ffe3521-d6f2-418a-81f5-d4f8aabcb1fd
date added to LUP
2019-02-01 12:18:29
date last changed
2024-03-19 00:24:13
@article{7ffe3521-d6f2-418a-81f5-d4f8aabcb1fd,
  abstract     = {{<p>We 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.</p>}},
  author       = {{Xu, Kerui and Clark, Charles P. and Poe, Brian L. and Lounsbury, Jenny A. and Nilsson, Johan and Laurell, Thomas and Landers, James P.}},
  issn         = {{0003-2700}},
  language     = {{eng}},
  month        = {{01}},
  number       = {{3}},
  pages        = {{2186--2191}},
  publisher    = {{The American Chemical Society (ACS)}},
  series       = {{Analytical Chemistry}},
  title        = {{Isolation of a Low Number of Sperm Cells from Female DNA in a Glass-PDMS-Glass Microchip via Bead-Assisted Acoustic Differential Extraction}},
  url          = {{http://dx.doi.org/10.1021/acs.analchem.8b04752}},
  doi          = {{10.1021/acs.analchem.8b04752}},
  volume       = {{91}},
  year         = {{2019}},
}