Skip to main content
SpringerLink
Log in

Microplate cell-retaining methodology for high-content analysis of individual non-adherent unanchored cells in a population

  • Published:
Biomedical Microdevices Aims and scope Submit manuscript

Abstract

A high throughput Microtiter plate Cell Retainer (MCR) has been developed to enable, for the first time, high-content, time-dependent analysis of the same single non-adherent and non-anchored cells in a large cell population, while bio-manipulating the cells. The identity of each cell in the investigated population is secured, even during bio-manipulation, by cell retention in a specially designed concave microlens, acting as a picoliter well (PW). The MCR technique combines micro-optical features and microtiter plate methodology. The array of PWs serves as the bottom of a microtiter plate, fitted with a unique flow damper element. The latter enables rapid fluid exchange without dislodging the cells from their original PWs, thus maintaining the cells' identity. Loading cell suspensions and reagents into the MCR is performed by simple pouring, followed by gravitational sedimentation and settling of cells into the PWs. Cell viability and cell division within the MCR were shown to be similar to those obtained under similar conditions in a standard microtiter plate. The efficiency of single cell occupancy in the MCR exceeded 90%. No cell dislodging was observed when comparing images before and after bio-manipulations (rinsing, staining, etc.). The MCR permits the performance of kinetic measurements on an individual cell basis. Data acquisition is governed by software, controlling microscope performance, stage position and image acquisition and analysis. The PW's unique micro-optical features enable rapid, simultaneous signal analysis of each individual cell, bypassing lengthy image analysis.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • I. Biran and D.R. Walt, Analytical Chemistry 74, 3046 (2002).

    Article  Google Scholar 

  • C.D.T. Bratten, P.H. Cobbold, and J.M. Cooper, Analytical Chemistry 69, 253 (1997).

    Article  Google Scholar 

  • D.M. Cannon, N. Winograd, and A.G. Ewing, Annual Review of Biophysics and Biomolecular Structure 29, 239 (2000).

    Article  Google Scholar 

  • S.J. Chen and S.J. Lillard, Analitical Chemistry 73, 111 (2001).

    Article  Google Scholar 

  • V.I. Chin, P. Taupin, S. Sanga, J. Scheel, F.H. Gage, and S.N. Bhatia, Biotechnology and Bioengineering 88, 399 (2004).

    Article  Google Scholar 

  • R. Clerval, M. Kühner, Z. Li, W. Minas, A. Fjällman, B. Witholt, and W.A. Duetz, Bioworld 6, 24 (2000).

    Google Scholar 

  • A. Colman-Lerner, A. Gordon, E. Serra, T. Chin, O. Resnekov, D. Endy, C.G. Pesce, and R. Brent, Nature 437, 699 (2005).

    Article  Google Scholar 

  • S.A. Connon and S.J. Giovannoni, Applied and Environmental Microbiology 68, 3878 (2002).

    Article  Google Scholar 

  • M. Deutsch and A. Weinreb, Cytometry 16, 214 (1994).

    Article  Google Scholar 

  • S.D. Doig, S.C.R. Pickering, G. Lye, and J.M. Woodley, Biotechnology and Bioengineering 80, 42 (2002).

    Article  Google Scholar 

  • W.D. Donachie and K.J. Begg, Nature 227, 1220 (1970).

    Article  Google Scholar 

  • W.A. Duetz, W. Minas, M. Kühner, R. Clerval, Z. Li, A.H.M. Fjällman, and B. Witholt, Bioworld 2, 8 (2001).

    Google Scholar 

  • J. Gao, X.F. Yin, and Z.L. Fang, Lab on a Chip 4, 47 (2004).

    Article  MATH  Google Scholar 

  • R.A. Flynn, A.L. Birkbeck, M. Gross, M. Ozkan, B. Shao, M.M. Wang, and S.C. Esener, Sensors and Actuators B Chemical 87, 239 (2002).

    Article  Google Scholar 

  • A. Gagro, S. Rabatic, I. Ivancic, K. Bendelja, J. Jelacic, A. Sabioncello, J. Misulic, D. Buneta, and D. Dekaris, Periodicum Biologorum 101, 17 (1999).

    Google Scholar 

  • Y. Huang and B. Rubinsky, Biomedical Microdevices 3, 145 (2000).

    MATH  Google Scholar 

  • I. Inoue, Y. Wakamoto, H. Moriguchi, K. Okanob, and K. Yasuda, Lab on a Chip 1, 50 (2001).

    Article  Google Scholar 

  • G.T. John, I. Klimant, C. Wittmann, and E. Heinzle, Biotechnology and Bioengineering 81, 829 (2003a).

    Article  Google Scholar 

  • G. John, D. Goelling, I. Klimant, H. Schneider, and E. Heinzle, Journal of Dairy Research 70, 327 (2003b)

    Article  Google Scholar 

  • S. Kumar, C. Wittmann, and E. Heinzle, Biotechnology Letters 26, 1 (2004).

    Article  Google Scholar 

  • P.C.H. Li, L. de Camprieu, J. Caia, and M. Sangarb, Lab on a Chip 4, 174 (2004).

    Article  Google Scholar 

  • A. Lueking, M. Horn, H. Eickhoff, K. Buessow, H. Lehrach, and G. Walter, Analytical Biochemistry 270, 103 (1999).

    Article  Google Scholar 

  • T.R. Malek, T.J. Fleming, and E.K.Codias, Seminars in Immunology 6, 105 (1994).

    Article  Google Scholar 

  • A. Malek and M.G. Khaledi, Analytical Biochemistry 268, 262 (1999).

    Article  Google Scholar 

  • F. Manca, Annali Dell Istituto Superiore Di Sanita 27, 15 (1991).

    Google Scholar 

  • R. Manns, MipTec-ICAR (1999). http://www.microplate.org/history/det_hist.htm

  • W. Minas, J.E. Bailey, and W.A. Antonie, van Leeuwenhoek 78, 297 (2000).

    Article  Google Scholar 

  • K.C. Neuman, E.H. Chadd, G.F. Liou, K. Bergman, and S.M. Block, Biophysical Journal 77, 2856 (1999).

    Google Scholar 

  • E. Ostuni, C.S. Chen, D.E. Ingber, and G.M. Whitesides, Langmuir, 17, 2828 (2001).

  • M. Ozkan, T .Pisanic, J. Scheel, C. Barlow, S. Esener, and S.N. Bhatia, Langmuir 19, 1532 (2003).

    Article  Google Scholar 

  • D.R. Plymale, J.R. Haskins, and F.A. de la Iglesia, Nature Medicine 5, 351 (1999).

    Article  Google Scholar 

  • W.J. Qian, C.A. Aspinwall, M.A. Battiste, and R.T. Kennedy, Analytical Chemistry 72, 711 (2000).

    Article  Google Scholar 

  • C. Sangdun, R.A. Creelman, J.E. Mullet, and R.A. Wing, Weeds World 2, 17 (1995).

    Google Scholar 

  • S. Santesson, M. Andersson, E. Degerman, T. Johansson, J. Nilsson, and S. Nilsson, Analytical Chemistry 72, 3412 (2000).

    Article  Google Scholar 

  • K.C. Schuster, I. Reese, E. Urlaub, J. R. Gapes, and B. Lendl, Analytical Chemistry 72, 5529 (2000).

    Article  Google Scholar 

  • J.A. Shapiro and C. Hsu, Journal of Bacteriology 171, 5963 (1989).

    Google Scholar 

  • H.M. Shapiro, Journal of Microbiological Methods 42, 3 (2000).

    Article  Google Scholar 

  • D.T. Stitt, M.S. Nagar, T.A. Haq, and M.R. Timmins, Biotechniques 32, 684 (2002).

    Google Scholar 

  • J. Tan, H. Shen, and W.M. Saltzman, Biophysical Journal 81, 2569 (2001).

    Article  Google Scholar 

  • D.L. Taylor, E.S. Woo, and K.A. Giuliano, Current Opinions in Biotechnology 12, 75 (2001).

    Article  Google Scholar 

  • G.E. Tsotsou, A.E.G. Cass, and G. Gianfranco, Biosensors and Bioelectronics 17, 119 (2002).

    Article  Google Scholar 

  • S. Weiss, G.T. John, I. Klimant, and E. Heinzle, Biotechnology Progress 18, 821 (2002).

    Article  Google Scholar 

  • G. Wennemuth, S. Eisoldt, H.P. Bode, H. Renneberg, P.J. Schiemann, and G. Aumuller, Andrologia 30, 141 (1998).

    Article  Google Scholar 

  • J. Voldman, M.L. Gray, M. Toner, and M.A. Schmidt, Analytical Chemistry 74, 3984 (2002).

    Article  Google Scholar 

  • E. S. Yeung, Analytical Chemistry 71, 522A (1999).

    Article  Google Scholar 

  • T. Zell, W.J. Kivens, S.A. Kellermann, and Y. Shimizu, Immunological Research 20, 127 (1999).

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. The Biophysical Interdisciplinary Jerome Schottenstein Center for the Research and the Technology of the Cellome, Department of Physics, Bar-Ilan University, Ramat Gan, 52900, Israel

    Assaf Deutsch, Naomi Zurgil, Ihar Hurevich, Yana Shafran, Elena Afrimzon, Pnina Lebovich & Mordechai Deutsch

Authors
  1. Assaf Deutsch
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Naomi Zurgil
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ihar Hurevich
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yana Shafran
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Elena Afrimzon
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Pnina Lebovich
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Mordechai Deutsch
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Mordechai Deutsch.

Electronic supplementary material

Supplementary video object (5,867 KB)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Deutsch, A., Zurgil, N., Hurevich, I. et al. Microplate cell-retaining methodology for high-content analysis of individual non-adherent unanchored cells in a population. Biomed Microdevices 8, 361–374 (2006). https://doi.org/10.1007/s10544-006-9143-y

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10544-006-9143-y

Keywords

Search

Navigation