Abstract
Cell-based arrays offer powerful tools for genomics/proteomics and drug discovery, and are widely applicable for most cell lines. However, it is challenging to apply cell-based arrays for in vitro diagnosis due to limited amount of patient samples. Here, we utilized and demonstrated microfluidic image cytometry (MIC), capable of quantitative, single-cell profiling of multiple signaling molecules using only 300–3,000 cells from clinical brain tumor specimens for in vitro molecular diagnosis. First, we characterized the PI3K/AKT/mTOR pathway, which is often over-activated in the brain tumors, in U87 brain tumor cell lines by measuring EGFR, PTEN, pAKT, and pS6 with a MIC platform, and applied this measurement to clinical brain tumor specimens. In conjunction with statistical analysis, we were able to characterize extensive inter- and intra-tumoral molecular heterogeneity.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Neumann, B., Held, M., Liebel, U., Erfle, H., Rogers, P., Pepperkok, R., Ellenberg, J. (2006) High-throughput RNAi screening by time-lapse imaging of live human cells. Nat Methods 3, 385–390.
Perlman, Z. E., Slack, M. D., Feng, Y., Mitchison, T. J., Wu, L. F., Altschuler, S. J. (2004) Multidimensional drug profiling by automated microscopy. Science 306, 1194–1198.
Tanaka, M., Bateman, R., Rauh, D., Vaisberg, E., Ramachandani, S., Zhang, C., Hansen, K. C., Burlingame, A. L., Trautman, J. K., Shokat, K. M., Adams, C. L. (2005) An unbiased cell morphology-based screen for new, biologically active small molecules. PLoS Biol 3, e128.
Pelkmans, L., Fava, E., Grabner, H., Hannus, M., Habermann, B., Krausz, E., Zerial, M. (2005) Nature 436, 78–86.
Kamei, K., Ohashi, M., Gschweng, E., Suh, J., Ho, Q., Yu, Z. T. F., Tang, J., Teitell, M., Clark, A. T., Pyle, A. D., Lee, K.-B., Witte, O. N., Tseng, H.-R. (2010) Microfluidic image cytometry for quantitative single-cell profiling of human pluripotent stem cells in chemically defined conditions. Lab on a Chip 10, 1113–1119
Sun, J., Masterman-Smith, M., Graham, N. A., Jiao, J., Mottahedeh, J., Ohashi, M., DeJesus, J., Laks, D. R., Panosyan, E., Park, J., Kamei, K., Lee, K.-B. Wang, H., Yu, Z. T.-F., Lu, Y.-T., Shuang, H., Li, K., Liu, M., Zhang, N., Wang, S., Angenieux, B., Samuels, E., Williams, D., Kankatit, V., Nathanson, D., van Dam, M., Phelps, M., Liau, L., Mischel, P. S., Lazareff, J. A., Kornblum, H. I., Wu, H., Yong, W. H., Graeber, T. G., Tseng, H.-R. (2010) A microfluidic platform for systems pathology: multiparameter single-cell signaling measurements of clinical brain tumor specimens. Cancer Res 70, 6128–6138.
Wang, M. Y., Lu, K. V., Zhu, S., Dia, E. Q., Vivanco, I., Shackleford, G. M., Cavenee, W. K., Mellinghoff, I. K., Cloughesy, T. F., Sawyers, C. L., Mischel, P. S (2006) Mammalian target of rapamycin inhibition promotes response to epidermal growth factor receptor kinase inhibitors in PTEN-deficient and PTEN-intact glioblastoma cells. Cancer Res 66, 7864–7869.
Brittain, S., Paul, K., Zhao, X. M., Whitesides, G. (1998) Soft lithography and microfabrication. Physics World 11, 31–36.
Whitesides, G. M., Ostuni, E., Takayama, S., Jiang, X., Ingber, D. E. (2001) Soft lithography in biology and biochemistry. Annu Rev Biomed Eng 3, 335–373.
Wehrens, R., Buydens, L. M. C. (2007) Self- and super-organizing maps in R: the kohonen package. J Stat Soft 21, 1–19.
Acknowledgments
This work was supported by the NanoSystems Biology Cancer Center (NSBCC), the Eli and Edythe Broad Center of Regenerative Medicine, and the Institute of Molecular Medicine at University of California, Los Angeles. H.R.T. was supported by California Institute of Regenerative Medicine. R.D. was supported by the Johnsson Comprehensive Cancer Center and the California NanoSystems Institute.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2011 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Kamei, Ki., Sun, J., Tseng, HR., Damoiseaux, R. (2011). Microfluidic Image Cytometry. In: Palmer, E. (eds) Cell-Based Microarrays. Methods in Molecular Biology, vol 706. Humana Press. https://doi.org/10.1007/978-1-61737-970-3_16
Download citation
DOI: https://doi.org/10.1007/978-1-61737-970-3_16
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61737-969-7
Online ISBN: 978-1-61737-970-3
eBook Packages: Springer Protocols