Springer Protocols: Abstract: Microchip-Based Enumeration of Human White Blood Cells

Microchip-Based Enumeration of Human White Blood Cells

By: Pierre N. Floriano², Shelley Acosta², Nicolaos Christodoulides², Shannon Weigum², John T. McDevitt²

Abstract

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The advent of flow cytometry has considerably changed the ways in which medical testing is conducted. However, the cost of flow cytometers, their large size, and their maintenance needs make them scarce in resource-poor settings and available almost only in clinical pathology laboratories in developed countries. Because cell enumeration is a basic and crucial support of diagnosis, prognosis, and treatment, an alternative cell-counting method that would potentially be cost-effective, portable, and suitable for use in resource-poor settings is warranted. We describe here a protocol for conducting cell-counting experiments in a simple microfluidic structure. This protocol describes how to build a simple microfluidic cell and perform a total white blood cell (WBC) count through capture and immunolabeling of the WBCs with an anti-CD45 antibody.

Affiliation(s): (2) Department of Chemistry and Biochemistry, University of Texas at Austin, Austin, TX

Book Title: Microchip-Based Assay Systems: Methods and Applications

Series: Methods in Molecular Biology | Volume: 385 | Pub. Date: Nov-13-2007 | Page Range: 53-64 | DOI: 10.1007/978-1-59745-426-1_5

Subject: Genetics/Genomics

Key Words: White blood cells - flow cytometry - microchip - CD45 - microfluidic sensor

References

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1.	Houwen, B. (2001) The differential cell count. Lab. Hematol. 7, 89–100.

2.	Hoffman, R. A., Maino, V. C., Recktenwald, D. J., and Webster, H. K. (2002) BD Biosciences contributions in CD4 counting and immune status for HIV/AIDS. Cytometry 50, 129–132.

3.	Rabellino, E. M., Forman, M. A., Howes, J. C. G., Mills, R. A., Quintana, J. A., Roth, P. A., and Wilkinson, J. G. (2002) Beckman Coulter and CD4+ T cells. Cytometry 50, 127–128.

4.	Brown, M., and Wittwer, C. (2000) Flow cytometry: principles and clinical applications in hematology. Clin. Chem. 46, 1221–1229.

5.	Cohen, J. (2004) Monitoring treatment: At what cost? Science 304, 1936–1936.

6.	Helbert, M. and Breuer, J. (2000) Monitoring patients with HIV disease. J. Clin. Pathol. 53(4), 266–272.

7.	Kvale, D., Aukrust, P., Osnes, K., Muller, F., and Froland, S. S. (1999) CD4+ and CD8+ lymphocytes and HIV RNA in HIV infection: high baseline counts and in particular rapid decrease of CD8+ lymphocytes predict AIDS. AIDS 13, 195–201.

8.	Laurence, J. (1993) Review: T-cell subsets in health, infectious disease, and idiopathic CD4+ T lymphocytopenia. Ann. Intern. Med. 119, 55–62.

9.	Janossy, G., Jani, I. V., Kahan, M., Barnett, D., Mandy, F., and Shapiro, H. (2002) Precise CD4 T-cell counting using red diode laser excitation: For richer, for poorer. Cytometry 50, 78–85.

10.	Earley, M. C., Vogt, R. F., Shapiro, H. M., et al. (2002) Report from a workshop on multianalyte microsphere assays. Cytometry 50, 239–242.

11.	Ahn, C. H., Choi, J. W., Beaucage, G., et al. (2004) Disposable Smart lab on a chip for point-of-care clinical diagnostics. Proc. IEEE 92, 154–173.

12.	Auroux, P. A., Iossifidis, D., Reyes, D. R., and Manz, A. (2002) Micro total analysis systems. 2. Analytical standard operations and applications. Anal. Chem. 74, 2637–2652.

13.	Broyles, B. S., Jacobson, S. C., and Ramsey, J. M. (2003) Sample filtration, concentration, and separation integrated on microfluidic devices. Anal. Chem. 75, 2761–2767.

14.	de Mello, A. J. and Beard, N. (2003) Dealing with ‘real’ samples: sample pretreatment in microfluidic systems. Lab on a Chip 3, 11N–19N.

15.	Landers, J. P. (2003) Molecular diagnostics on electrophoretic microchips. Anal. Chem. 75, 2919–2927.

16.	Liu, J., Hansen, C., and Quake, S. R. (2003) Solving the “world-to-chip” interface problem with a microfluidic matrix. Anal. Chem. 75, 4718–4723.

17.	Manz, A., and Eijkel, J. C. T. (2001) Miniaturization and chip technology. What can we expect? Pure Appl. Chem. 73, 1555–1561.

18.	Manz, A., Graber, N., and Widmer, H. M. (1990) Miniaturized total chemicalanalysis systems—a novel concept for chemical sensing. Sensors Actuators B-Chem. 1, 244–248.

19.	McCarley, R. L., Vaidya, B., Wei, S. Y., et al. (2005) Resist-free patterning of surface architectures in polymer-based microanalytical devices. J. Am. Chem. Soc. 127, 842–843.

20.	McClain, M. A., Culbertson, C. T., Jacobson, S. C., and Ramsey, J. M. (2001) Flow cytometry of Escherichia coli on nucrifluidic devices. Anal. Chem. 73, 5334–5338.

21.	McDonald, J. C. and Whitesides, G. M. (2002) Poly(dimethylsiloxane) as a material for fabricating microfluidic devices. Accounts Chem. Res. 35, 491–499.

22.	Minc, N. and Viovy, J. L. (2004) Microfluidics and biological applications: the stakes and trends. Comptes Rendus Physique 5, 565–575.

23.	Pamme, N., Koyama, R., and Manz, A. (2003) Counting and sizing of particles and particle agglomerates in a microfluidic device using laser light scattering: application to a particle-enhanced immunoassay. Lab on a Chip 3, 187–192.

24.	Sia, S. K. and Whitesides, G. M. (2003) Microfluidic devices fabricated in poly(dimethylsiloxane) for biological studies. Electrophoresis 24, 3563–3576.

25.	Situma, C., Wang, Y., Hupert, M., Barany, F., McCarley, R. L., and Soper, S. A. (2005) Fabrication of DNA microarrays onto poly(methyl methacrylate) with ultraviolet patterning and microfluidics for the detection of low-abundant point mutations. Anal. Biochem. 340, 123–135.

26.	Tudos, A. J., Besselink, G. A. J., and Schasfoort, R. B. M. (2001) Trends in miniaturized total analysis systems for point-of-care testing in clinical chemistry. Lab on a Chip 1, 83–95.

27.	Verpoorte, E. (2003) Beads and chips: new recipes for analysis. Lab on a Chip 3, 60N–68N.

28.	Verpoorte, E. and De Rooij, N. F. (2003) Microfluidics meets MEMS. Proc. IEEE 91, 930–953.

29.	Vilkner, T., Janasek, D., and Manz, A. (2004) Micro total analysis systems. Recent developments. Anal. Chem. 76, 3373–3385.

30.	Rodriguez, W. R., Christodoulides, N., Floriano, P. N., et al. (2005) A microchip CD4 counting method for HIV monitoring in resource-poor settings. Plos Med. 2, 663–672.

31.	Rasband, W. S. (2006) Image, U.S., National Institutes of Health, Bethesda, MD, U.S.A., http://rsb.info.mih.gov/ij/,1997–2006.

32.	Piruska, A., Nikcevic, I., Lee, S. H., et al. (2005) The autofluorescence of plastic materials and chips measured under laser irradiation. Lab on a Chip 5, 1348–1354.

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