Annals of Biomedical Engineering

, Volume 33, Issue 2, pp 121–130 | Cite as

Laser Printing of Single Cells: Statistical Analysis, Cell Viability, and Stress

  • Jason A. Barron
  • David B. Krizman
  • Bradley R. Ringeisen
Article

Abstract

Methods to print patterns of mammalian cells to various substrates with high resolution offer unique possibilities to contribute to a wide range of fields including tissue engineering, cell separation, and functional genomics. This manuscript details experiments demonstrating that BioLP TM Biological Laser Printing, can be used to rapidly and accurately print patterns of single cells in a noncontact manner. Human osteosarcoma cells were deposited into a biopolymer matrix, and after 6 days of incubation, the printed cells are shown to be 100% viable. Printing low numbers of cells per spot by BioLPTM is shown to follow a Poisson distribution, indicating that the reproducibility for the number of cells per spot is therefore determined not by the variance in printed volume per drop but by random sampling statistics. Potential cell damage during the laser printing process is also investigated via immunocytochemical studies that demonstrate minimal expression of heat shock proteins by printed cells. Overall, we find that BioLPTM is able to print patterns of osteosarcoma cells with high viability, little to no heat or shear damage to the cells, and at the ultimate single cell resolution.

Keywords

BioLPTM Laser-induced forward transfer Tissue engineering Cell separation Cell seeding 

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References

  1. 1.
    Azuma, N., N. Akasaka, H. Kito, M. Ikeda, V. Gahtan, T. Sasjima, and B. E. Sumpio. Role of p38 MAP kinase in endothelial cell alignment induced by fluid shear stress. Am. J. Physiol. Heart Circ. Physiol. 280:H189–H197, 2001.Google Scholar
  2. 2.
    Bancroft, G. N., V. I. Sikavitsas, J. V. D. Dolder, T. L. Sheffield, C. G. Ambrose, J. A. Jansen, and A. G. Mikos. Fluid flow increases mineralization matrix deposition in 3D perfusion culture of marrow stromal osteoblasts in a dose-dependent manner Proc. Nat. Acad. Sci. 99:12600–12604, 2002CrossRefGoogle Scholar
  3. 3.
    Barron, J. A., P. K. Wu, H. D. Ladouceur, and B. R. Ringeisen. Biological laser printing: A novel technique for creating heterogeneous three-dimensional cell patterns Biomed. Microdev. 6:139–147, 2004CrossRefGoogle Scholar
  4. 4.
    Barron, J. A., B. J. Spargo, and B. R. Ringeisen. Biological laser printing of three-dimensional cellular structures. App. Phys. A 79:1027–1030, 2004CrossRefGoogle Scholar
  5. 5.
    Barron, J. A., R. Rosen, J. Jones-Meehan, B. J. Spargo, S. Belkin, and B. R. Ringeisen. Biological laser printing of genetically modified Escherichia coli for biosensor applications. Biosens. Bioelectr. 20:246–252, 2004CrossRefGoogle Scholar
  6. 6.
    Buican, T. N., M. J. Smyth, H. A. Crissman, G. C. Salzman, C. C. Stewart, and J. C. Martin. Automated single-cell manipulation and sorting by light trapping. App. Optics 26:5311–5316, 1987.CrossRefGoogle Scholar
  7. 7.
    Chrisey, D. B., R. A. McGill, J. S. Horwitz, A. Pique, B. R. Ringeisen, D. M. Bubb, and P. K. Wu. Laser deposition of polymer and biomaterial films. Chem. Rev. 103:553–576, 2003. CrossRefGoogle Scholar
  8. 8.
    Darzynkiewicz, Z., E. Bedner, X. Li, W. Gorczyca, and M. R. Melamed. Laser-scanning cytometry: A new instrumentation with many applications. Exp. Cell Res. 249:1–12, 1999CrossRefGoogle Scholar
  9. 9.
    Davey, H. M., and D. B. Kell. Flow cytometry and cell sorting of heterogeneous microbial populations: The importance of single-cell analyses. Microbiol. Rev. 60:641–661, 1996.Google Scholar
  10. 10.
    Freed, L. E., and G. Vunjak-Novakovic. Tissue culture bioreactors: Chondrogenesis as a model system. In: Principles of Tissue Engineering, edited by R. P. Lanza, W. Chick, and R. Langer. Austin: Landes, 1997, pp. 151–165.Google Scholar
  11. 11.
    Grover, S. C., A. G. Skirtach, R. C. Gauthier, and C. P. Grover. Automated single-cell sorting system based on optical trapping. J. Biomed. Optics 6:14–22, 2001.CrossRefGoogle Scholar
  12. 12.
    Ikeda, K., V. P. Michelangeli, T. J. Martin, and D. M. Findlay. Type-I collagen substrate increases calcitonin and parathyroid-hormone receptor-mediated signal-transduction in UMR 106-06 osteoblast-like cells. J. Cell. Physiol. 156:130–137, 1993.Google Scholar
  13. 13.
    Jager, E. W. H., C. Immerstrand, K. H. Peterson, K. E. Magnusson, I. Lundstrom, and O. Inganas. The cell clinic: Closable microvials for single cell studies. Biomed. Micro. 4:177–187, 2002CrossRefGoogle Scholar
  14. 14.
    Jakab, K., A. Neagu, V. Mironov, R. R. Markwald, and G. Forgacs. Engineering biological structures of prescribed shape using self-assembling multicellular systems. Proc. Nat. Acad. Sci. 101:2864–2869, 2004CrossRefGoogle Scholar
  15. 15.
    Karaiskou, A., I. Zergioti, C. Fotakis, M. Kapsetaki, and D. Kafetzopoulos. Microfabrication of biomaterials by the sub-ps laser-induced forward transfer process. Appl. Surf. Sci. 208–209:245–249, 2003.CrossRefGoogle Scholar
  16. 16.
    Kasili, R. M., B. M. Cullum, G. D. Griffin, and T. Vo-Dinh. Nanosensor for in vivo measurement of the carcinogen benzo[a]pyrene in a single cell. J. Nanosci. Nanotech. 2:653–658, 2002.CrossRefGoogle Scholar
  17. 17.
    Katsuragi, T., and Y. Tani. Single-cell sorting of microorganisms by flow or slide-based (including laser scanning) cytometry. Acta Biotechnol. 21:99–115, 2001CrossRefGoogle Scholar
  18. 18.
    Langer, R., and J. P. Vacanti. Tissue engineering. Science 260:920, 1993.PubMedGoogle Scholar
  19. 19.
    Mather, J. P., and P. E. Roberts. Introduction to Cell and Tissue Culture. New York: Plenum Press, 1998, pp. 66–70.Google Scholar
  20. 20.
    Meyer, S. L. Data Analysis for Scientists and Engineers. New York: Wiley, 1975, pp. 202–222.Google Scholar
  21. 21.
    Mironov, V., T. Boland, T. Trusk, G. Forgacs, and R. R. Markwald. Organ printing: Computer-aided jet-based 3D tissue engineering. Trends Biotechnol. 21:157–161, 2003CrossRefGoogle Scholar
  22. 22.
    Müller, T., G. Gradl, S. Howitz, S. Shirley, T. Schnelle, and G. Fuhr. A 3-D microelectrode system for handling and caging single cells and particles. Biosens. Bioelectron. 14:247–256, 1999CrossRefGoogle Scholar
  23. 23.
    Nerem, R. M. Cellular engineering. Ann. Biomed. Eng. 19:529, 1991.Google Scholar
  24. 24.
    Nishizuka, S., L. Charboneau, L. Young, S. Major, W. C. Renhold, M. Waltham, H. Kouros-Mehr, K. J. Bussey, J. K. Lee, V. Espina, P. J. Munson, E. Petricoin III, L. A. Liotta, and J. N. Weinstein. Proteomic profiling of the NCI-60 cancer cell lines using new high-density reverse-phase lysate microarrays. Proc. Nat. Acad. Sci. 100:14229–14234, 2003CrossRefGoogle Scholar
  25. 25.
    Odde, D. J., and M. J. Renn. Laser-guided direct writing for applications in biotechnology. Trends Biotechnol. 17:385–389, 2000.CrossRefGoogle Scholar
  26. 26.
    Oishi, Y., K. Taniguchi, H. Matsumoto, A. Ishihara, Y. Ohira, and R. R. Roy. Differential responses of HSPs to heat stress in slow and fast regions of rat gastrocnemius muscle. Muscle Nerve 28:587–94, 2003CrossRefGoogle Scholar
  27. 27.
    Reichle, C., K. Sparbier, T. Müller, T. Schnelle, P. Walden, and G. Fuhr. Combined laser tweezers and dielectric field cage for the analysis of receptor-ligand interactions on single cells. Electrophoresis, 22:272–282, 2001CrossRefGoogle Scholar
  28. 28.
    Ringeisen, B. R., D. B. Chrisey, A. Piqué, R. Modi, D. Young, M. Bucaro, J. Jones-Meehan, and B. J. Spargo. Generation of mesoscopic patterns of viable Escherichia coli by ambient laser transfer. Biomaterials 23:161–166, 2002.CrossRefGoogle Scholar
  29. 29.
    Ringeisen, B. R., H. Kim, J. A. Barron, D. B. Krizman, D. B. Chrisey, S. Jackman, R. Y. C. Auyeung, and B. J. Spargo. Laser printing of pluripotent embryonal carcinoma cells. Tissue Eng. 10: 483–491, 2004.CrossRefGoogle Scholar
  30. 30.
    Ringeisen, B. R., P. K. Wu, H. Kim, A. Piqué, R. Y. C. Auyeung, D. Young, D. B. Chrisey, and D. B. Krizman. Picoliter-scale protein microarrays by laser direct write. Biotech. Prog. 18:1126–1129, 2002CrossRefGoogle Scholar
  31. 31.
    Rudensky, B., E. Paz, G. Altarescu, D. Raveh, E. Elstein, and A. Zimran. Fluorescent flow cytometric assay: A new diagnostic tool for measuring beta-glucocerebrosidase activity in Gaucher disease. Blood Cell Mol. Dis. 1:97–99, 2003.CrossRefGoogle Scholar
  32. 32.
    Shapiro, H. H. Practical Flow Cytometry. New York: Wiley-Liss, 1995.Google Scholar
  33. 33.
    Tazi, K. A., E. Barriere, R. Moreau, J. Heller, P. Sogni, D. Pateron, O. Poirel, and D. Lebrec. Role of shear stress in aortic eNOS up-regulation in rats with biliary cirrhosis. Gastroenterology 122:1869–1877, 2002Google Scholar
  34. 34.
    Weaver, J. L. Introduction to flow cytometry. Methods 21:199–201, 2000CrossRefGoogle Scholar
  35. 35.
    Wu, P. K., B. R. Ringeisen, J. Callahan, M. Brooks, D. M. Bubb, H. D. Young, A. Piqué, B. J. Spargo, R. A. McGill, and D. B. Chrisey. The deposition, structure, pattern deposition, and activity of biomaterial thin-films by matrix-assisted pulsed-laser evaporation (MAPLE) and MAPLE direct write. Thin Solid Films 398:607, 2001.CrossRefGoogle Scholar
  36. 36.
    Wu, P. K., B. R. Ringeisen, D. B. Krizman, S. M. Hewitt, C. G. Frondoza, M. Brooks, D. M. Bubb, R. C. Y. Auyeung, H. Kim, A. Pique, J. M. Fitz-Gerald, B. J. Spargo, R. A. McGill, and D. B. Chrisey. Laser transfer of biomaterials: Matrix-assisted pulsed laser evaporation (MAPLE) and MAPLE Direct Write. Rev. Sci. Instr. 74:2546–2557, 2003CrossRefGoogle Scholar
  37. 37.
    Yoshida, S., E. Shimizu, T. Ogura, M. Takada, and S. Sone. Stimulatory effect of reconstituted basement membrane components (Matrigel) on the colony formation of a panel of human l ung cancer cell lines in soft agar. J. Cancer Res. Clin. Oncol. 123:301–309, 1997CrossRefGoogle Scholar

Copyright information

© Biomedical Engineering Society 2005

Authors and Affiliations

  • Jason A. Barron
    • 1
  • David B. Krizman
    • 1
  • Bradley R. Ringeisen
    • 1
    • 2
  1. 1.Chemical Dynamics and Diagnostics Branch, Chemistry DivisionNaval Research Laboratory
  2. 2.Biological Chemistry Branch, Code 6113

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