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Concurrent Isolation of Lymphocytes and Granulocytes Using Prefocused Free Flow Acoustophoresis

Grenvall, Carl LU ; Magnusson, Cecilia LU ; Lilja, Hans LU and Laurell, Thomas LU (2015) In Analytical Chemistry 87(11). p.5596-5604
Abstract
Microchip-based free flow acoustophoresis (FFA) in combination with two-dimensional cell prefocusing enables concurrent multiple target outlet fractionation of leukocytes into subpopulations (lymphocytes, monocytes, and granulocytes); we report on this method here. We also observed significantly increased accuracy in size-based fractionation of microbeads as compared to previously presented FFA multiple outlet systems. Fluorescence microscopy illustrates the importance of two-dimensional prefocusing where a sample mixture of 3, 7, and 10 mu m beads are separated into well-confined particle streams and collected in their respective target outlets. Flow cytometry data for lymphocytes and granulocytes, respectively, in their corresponding... (More)
Microchip-based free flow acoustophoresis (FFA) in combination with two-dimensional cell prefocusing enables concurrent multiple target outlet fractionation of leukocytes into subpopulations (lymphocytes, monocytes, and granulocytes); we report on this method here. We also observed significantly increased accuracy in size-based fractionation of microbeads as compared to previously presented FFA multiple outlet systems. Fluorescence microscopy illustrates the importance of two-dimensional prefocusing where a sample mixture of 3, 7, and 10 mu m beads are separated into well-confined particle streams and collected in their respective target outlets. Flow cytometry data for lymphocytes and granulocytes, respectively, in their corresponding outlets verify concurrent isolation of leukocyte subpopulations with high purity (95.2 +/- 0.6% and 98.5 +/- 0.7%) and high recovery (86.5 +/- 10.9% and 68.4 +/- 10.6%). A relatively low purity and high recovery of monocytes (25.2% +/- 5.4% and 83.1 +/- 4.3%) was obtained in the third target outlet. No subpopulation bias was observed. These data demonstrate an unprecedented separation of leukocyte subpopulations at flow rates of similar to 100 mu L/min and similar to 1 M cells/mL sample concentrations, not previously reported in acoustofluidic systems. Two-dimensional prefocusing FFA with multiple target outlets is a viable alternative to current methods for particle fractionation and cell isolation, requiring a minimum of sample preparation and lowering analysis time and cost. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Analytical Chemistry
volume
87
issue
11
pages
5596 - 5604
publisher
The American Chemical Society
external identifiers
  • wos:000355779500023
  • scopus:84930635154
ISSN
1520-6882
DOI
10.1021/acs.analchem.5b00370
language
English
LU publication?
yes
id
5aecc78e-10a8-4a8d-a992-f32f2a699c72 (old id 7605061)
date added to LUP
2015-07-27 14:13:38
date last changed
2017-12-10 03:05:50
@article{5aecc78e-10a8-4a8d-a992-f32f2a699c72,
  abstract     = {Microchip-based free flow acoustophoresis (FFA) in combination with two-dimensional cell prefocusing enables concurrent multiple target outlet fractionation of leukocytes into subpopulations (lymphocytes, monocytes, and granulocytes); we report on this method here. We also observed significantly increased accuracy in size-based fractionation of microbeads as compared to previously presented FFA multiple outlet systems. Fluorescence microscopy illustrates the importance of two-dimensional prefocusing where a sample mixture of 3, 7, and 10 mu m beads are separated into well-confined particle streams and collected in their respective target outlets. Flow cytometry data for lymphocytes and granulocytes, respectively, in their corresponding outlets verify concurrent isolation of leukocyte subpopulations with high purity (95.2 +/- 0.6% and 98.5 +/- 0.7%) and high recovery (86.5 +/- 10.9% and 68.4 +/- 10.6%). A relatively low purity and high recovery of monocytes (25.2% +/- 5.4% and 83.1 +/- 4.3%) was obtained in the third target outlet. No subpopulation bias was observed. These data demonstrate an unprecedented separation of leukocyte subpopulations at flow rates of similar to 100 mu L/min and similar to 1 M cells/mL sample concentrations, not previously reported in acoustofluidic systems. Two-dimensional prefocusing FFA with multiple target outlets is a viable alternative to current methods for particle fractionation and cell isolation, requiring a minimum of sample preparation and lowering analysis time and cost.},
  author       = {Grenvall, Carl and Magnusson, Cecilia and Lilja, Hans and Laurell, Thomas},
  issn         = {1520-6882},
  language     = {eng},
  number       = {11},
  pages        = {5596--5604},
  publisher    = {The American Chemical Society},
  series       = {Analytical Chemistry},
  title        = {Concurrent Isolation of Lymphocytes and Granulocytes Using Prefocused Free Flow Acoustophoresis},
  url          = {http://dx.doi.org/10.1021/acs.analchem.5b00370},
  volume       = {87},
  year         = {2015},
}