Abstract
We report a rapid, low-cost, portable microfluidic sedimentation cytometer (SeCy) for assessing the somatic cell count and fat content of milk in 15 min using a “sample-in, answer-out” approach. The system consists of 12 independent microfluidic devices, essentially flattened funnel structures, fabricated on the footprint of a single plastic compact disc (CD). Each funnel structure holds 150 μL of milk, has an inlet for milk filling and an outlet for air to escape, and ends in a narrow, closed-end microfluidic channel that facilitates packing of the cells into a column whose length is proportional to cell count. The closed-end channel provides accurate cell counts over the range 50,000–>3,000,000 cells per mL. The assay separates cells and fat globules based on their densities (by differential sedimentation), concentrating white cells in the closed-end channel near the outer rim of the CD for estimation of total “cell pellet” volume, while fat globules move toward the center of disc rotation, forming a fat “band” in the funnel. After adding milk to two or more microfluidic devices, the CD is loaded onto a custom-built reader unit that spins the disc for 15 min. Two low-cost microscopes in the reader image the centrifuged cell pellet and the fat band, providing a sufficiently accurate cell count to diagnose mastitis and measuring fat content as an indication of health and nutritional status.
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J.R. Ashes, S.K. Gulati, T.W. Scott, J. Dairy Sci. 80(9), 2204–2212 (1997)
D.A. Ateya, J.S. Erickson, P.B. Howell, L.R. Hilliard, J.P. Golden, F.S. Ligler, Anal. Bioanal. Chem. 391(5), 1485–1498 (2008)
E. Berry, N. Middleton, M. Gravenor, J.E. Hillerton, Science (or art) of cell counting. Proceedings of the British Mastisis Conference, (2003)
X. Cheng, Y.S. Liu, D. Irimia, U. Demirci, L.J. Yang, L. Zamir, W.R. Rodriguez, M. Toner, R. Bashir, Lab Chip 7(6), 746–755 (2007a)
X.H. Cheng, D. Irimia, M. Dixon, K. Sekine, U. Demirci, L. Zamir, R.G. Tompkins, W. Rodriguez, M. Toner, Lab Chip 7(2), 170–178 (2007b)
X.H. Cheng, A. Gupta, C.C. Chen, R.G. Tompkins, W. Rodriguez, M. Toner, Lab Chip 9(10), 1357–1364 (2009)
T.D. Chung, H.C. Kim, Electrophoresis 28(24), 4511–4520 (2007)
J.A. Davis, D.W. Inglis, K.J. Morton, D.A. Lawrence, L.R. Huang, S.Y. Chou, J.C. Sturm, R.H. Austin, Proc. Natl. Acad. Sci. U. S. A. 103(40), 14779–84 (2006)
A. Donev, I. Cisse, D. Sachs, E. Variano, F.H. Stillinger, R. Connelly, S. Torquato, P.M. Chaikin, Science 303(5660), 990–993 (2004)
D.O. Forcato, M.P. Carmine, G.E. Echeverria, R.P. Pecora, S.C. Kivatinitz, J. Dairy Sci. 88(2), 478–481 (2005)
P.F. Fox, P.L.H. McSweeney, Advanced Dairy Chemistry: Lipids, vol. 2 (Springer, USA, 2003)
D. Fromer, D. Lerche, Arc. Appl. Mech. 72(2–3), 85–95 (2002)
J. Garcia-Cordero, L. Kent, I. Dimov, C. Viguier, L. Lee, A. Ricco, Microfluidic CD-Based Somatic Cell Counter for the Early Detection of Bovine Mastitis. 12th International Conference on Miniaturized Systems for Chemistry and Life Sciences, microTAS, San Diego (2008)
C. Gonzalo, J.C. Boixo, J.A. Carriedo, F. San Primitivo, J. Dairy Sci. 87(11), 3623–3628 (2004)
R.J. Harmon, J. Dairy Sci. 77(7), 2103–2112 (1994)
J.E. Hillerton, E.A. Berry, Quality of the milk supply: Europe and regulations versus practice, National Mastitis Council Annual Meeting Proceedings 207–214 (2004)
J.E. Hillerton, E.A. Berry, J. Appl, Microbiol. 98(6), 1250–1255 (2005)
D. Huh, W. Gu, Y. Kamotani, J.B. Grotberg, S. Takayama, Physiol. Meas. 26(3), R73–R98 (2005)
K. Huijps, T.J.G.M. Lam, H. Hogeveen, J. Dairy Res. 75(1), 113–120 (2008)
D.W. Inglis, K.J. Morton, J.A. Davis, T.J. Zieziulewicz, D.A. Lawrence, R.H. Austin, J.C. Sturm, Lab Chip 8(6), 925–31 (2008)
M.E. Kehrli, D.E. Shuster, J. Dairy Sci. 77(2), 619–627 (1994)
D.M. Leatzow, B.J. Van Wie, B.N. Weyrauch, T.O. Tiffany, Anal. Chim. Acta 435(2), 299–307 (2001)
C.S. Lee, F.B.P. Wooding, P. Kemp, J. Dairy Res. 47(1), 39–50 (1980)
K.E. Leslie, J.T. Jansen, G.H. Lim, Opportunities and implications for improved on-farm cowside diagnostics. Proc. DeLaval Hygiene Symp. (2002)
J.S. Moon, H.C. Koo, Y.S. Joo, S.H. Jeon, D.S. Hur, C.I. Chung, H.S. Jo, Y.H. Park, J. Dairy Sci. 90(5), 2253–2259 (2007)
C. Nightingale, K. Dhuyvetter, R. Mitchell, Y. Schukken, J. Dairy Sci. 91(3), 1236–1244 (2008)
S. Pyorala, Vet. Res. 34(5), 565–578 (2003)
Y.H. Schukken, D.J. Wilson, F. Welcome, L. Garrison-Tikofsky, R.N. Gonzalez, Vet. Res. 34(5), 579–596 (2003)
P. Sethu, A. Sin, M. Toner, Lab Chip 6(1), 83–89 (2006)
C. Viguier, S. Arora, N. Gilmartin, K. Welbeck, R. O’Kennedy, Trends Biotechnol. 27(8), 486–493 (2009)
P. Walstra, in Advanced Dairy Chemistry, ed. by P.F. Fox (Chapman & Hall, London, 1995), pp. 131–151
P. Walstra, T.J. Geurts, A. Noomen, A. Jellema, M.A.J.S. van Boekel, Dairy Technology: Principles of Milk Properties and Processes (Marcel Dekker, NY, USA, 1999)
J.J. Windig, M.P. Calus, R.F. Veerkamp, in Mastitis in Dairy Production: Current Knowledge and Future Solutions, ed. by H. Hogeveen (Wageningen Academic Publishers, The Netherlands, 2005), pp. 254–259
Acknowledgements
This work was supported by Science Foundation Ireland under Grant No. 05/CE3/B754. We are grateful to Jim Flynn and Brendan Kavanagh in Teagasc, MoorePark for providing and analyzing milk samples in the MilkoScan system. We also acknowledge Lorcan Kent for help with SEM images and Caroline Viguier for help during initial experiments. We thank Prof. Luke P. Lee for helpful discussions and Ivan Dimov for helping with image analysis. We also thank Dr. Holger Becker of Microfluidic ChipShop for helpful discussions in adapting our CD design to the injection molding process.
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Garcia-Cordero, J.L., Barrett, L.M., O’Kennedy, R. et al. Microfluidic sedimentation cytometer for milk quality and bovine mastitis monitoring. Biomed Microdevices 12, 1051–1059 (2010). https://doi.org/10.1007/s10544-010-9459-5
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DOI: https://doi.org/10.1007/s10544-010-9459-5