A continuous-flow, microfluidic fraction collection device.

Christopher A Baker, Michael G Roper
Author Information
  1. Christopher A Baker: Department of Chemistry and Biochemistry, Florida State University, Tallahassee, 32306, USA.

Abstract

A microfluidic device is presented that performs electrophoretic separation coupled with fraction collection. Effluent from the 3.5 cm separation channel was focused via two sheath flow channels into one of seven collection channels. By holding the collection channels at ground potential and varying the voltage ratio at the two sheath flow channels, the separation effluent was directed to either specific collection channels, or could be swept past all channels in a defined time period. As the sum of the voltages applied to the two sheath flow channels was constant, the electric field remained at 275 V/cm during the separation regardless of the collection channel used. The constant potential in the separation channel allowed uninterrupted separation for late-migrating peaks while early-migrating peaks were being collected. To minimize the potential for carryover between fractions, the device geometry was optimized using a three-level factorial model. The optimum conditions were a 22.5° angle between the sheath flow channels and the separation channel, and a 350 μm length of channel between the separation outlet and the fraction channels. Using these optimized dimensions, the device performance was evaluated by separation and fraction collection of a fluorescently-labeled amino acid mixture. The ability to fraction collect on a microfluidic platform will be especially useful during automated or continuous operation of these devices or to collect precious samples.

Keywords

References

  1. Electrophoresis. 2003 Nov;24(21):3745-7 [PMID: 14613200]
  2. Anal Chem. 1997 Jul 1;69(13):2617 [PMID: 21639399]
  3. Anal Chem. 2010 Feb 15;82(4):1253-60 [PMID: 20092256]
  4. Electrophoresis. 2007 Dec;28(24):4661-7 [PMID: 18004713]
  5. Electrophoresis. 2009 Dec;30(24):4277-84 [PMID: 20013913]
  6. Science. 1988 Oct 28;242(4878):562-4 [PMID: 3140381]
  7. Anal Bioanal Chem. 2009 May;394(1):187-98 [PMID: 19290514]
  8. Anal Chem. 2009 Aug 15;81(16):6837-42 [PMID: 19621896]
  9. Anal Chem. 2010 Feb 1;82(3):967-73 [PMID: 20058879]
  10. Small. 2009 Dec;5(24):2828-34 [PMID: 19787681]
  11. Proc Natl Acad Sci U S A. 2006 Dec 19;103(51):19272-7 [PMID: 17159153]
  12. Anal Chem. 2009 Feb 15;81(4):1477-81 [PMID: 19152307]
  13. Anal Chem. 2004 Nov 1;76(21):6419-25 [PMID: 15516136]
  14. J Chromatogr A. 2003 Aug 29;1010(2):255-68 [PMID: 12974295]
  15. Lab Chip. 2008 May;8(5):801-9 [PMID: 18432352]
  16. Anal Chem. 2007 Oct 1;79(19):7485-91 [PMID: 17718538]
  17. Anal Chem. 2007 Nov 15;79(22):8557-63 [PMID: 17953452]
  18. Anal Chem. 2002 Apr 1;74(7):1737-40 [PMID: 12033268]

Grants

  1. R01 DK080714/NIDDK NIH HHS
  2. R01 DK080714-04/NIDDK NIH HHS

MeSH Term

Amino Acids
Electrophoresis
Equipment Design
Fluorescein-5-isothiocyanate
Microfluidic Analytical Techniques
Spectrometry, Fluorescence

Chemicals

Amino Acids
Fluorescein-5-isothiocyanate

Word Cloud

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