Skip to main content
SpringerLink
Log in

Single-Cell Microinjection Technologies

  • Protocol
  • First Online:
Single-Cell Analysis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 853))

Abstract

Single-cell microinjection, as a mechanical delivery tool, has been used for transferring substances into transfection or infection challenging cells. Here, we discuss the advantages and applications of microinjection, list the materials needed for performing microinjection experiments, and describe the methods of single-cell microinjection into suspended and attached cells.

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

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Zhang, Y., and Yu, L. C. (2008) Microinjection as a tool of mechanical delivery, Curr Opin Biotechnol 19, 506–510.

    Google Scholar 

  2. Zhang, Y., and Yu, L. C. (2008) Single-cell microinjection technology in cell biology, Bioessays 30, 606–610.

    Google Scholar 

  3. Zhang, Y., and LeBlanc, A. (2002) Microinjection of proteins, peptides and genes in human neuronal apoptosis study., Vol. 37, 2 ed., Humana Press, Totowa, New Jersey,.

    Google Scholar 

  4. Zhang, Y., Champagne, N., Beitel, L. K., Goodyer, C. G., Trifiro, M., and LeBlanc, A. (2004) Estrogen and androgen protection of human neurons against intracellular amyloid beta1-42 toxicity through heat shock protein 70, J Neurosci 24, 5315–5321.

    Google Scholar 

  5. Zhang, Y., Goodyer, C., and LeBlanc, A. (2000) Selective and protracted apoptosis in human primary neurons microinjected with active caspase-3, -6, -7, and −8, J Neurosci 20, 8384–8389.

    Google Scholar 

  6. Zhang, Y., Hong, Y., Bounhar, Y., Blacker, M., Roucou, X., Tounekti, O., Vereker, E., Bowers, W. J., Federoff, H. J., Goodyer, C. G., and LeBlanc, A. (2003) p75 neurotrophin receptor protects primary cultures of human neurons against extracellular amyloid beta peptide cytotoxicity, J Neurosci 23, 7385–7394.

    Google Scholar 

  7. Zhang, Y., McLaughlin, R., Goodyer, C., and LeBlanc, A. (2002) Selective cytotoxicity of intracellular amyloid beta peptide1-42 through p53 and Bax in cultured primary human neurons, J Cell Biol 156, 519–529.

    Google Scholar 

  8. Zhang, Y., Tounekti, O., Akerman, B., Goodyer, C. G., and LeBlanc, A. (2001) 17-beta-estradiol induces an inhibitor of active caspases, J Neurosci 21, RC176.

    Google Scholar 

  9. Kacharmina, J. E., Crino, P. B., and Eberwine, J. (1999) Preparation of cDNA from single cells and subcellular regions, Methods Enzymol 303, 3–18.

    Google Scholar 

  10. Malter, H. E. (1992) Early development of micromanipulation. In Micromanipulation of Human Gametes and Embryos, Raven Press, NY.

    Google Scholar 

  11. Wadsworth, P. (1999) Microinjection of mitotic cells, Methods Cell Biol 61, 219–231.

    Google Scholar 

  12. Leonetti, J. P., Mechti, N., Degols, G., Gagnor, C., and Lebleu, B. (1991) Intracellular distribution of microinjected antisense oligonucleotides, Proc Natl Acad Sci USA 88, 2702–2706.

    Google Scholar 

  13. Romo, X., Pasten, P., Martinez, S., Soto, X., Lara, P., de Arellano, A. R., Torrejon, M., Montecino, M., Hinrichs, M. V., and Olate, J. (2007) xRic-8 is a GEF for Gsalpha and participates in maintaining meiotic arrest in Xenopus laevis oocytes, J Cell Physiol.

    Google Scholar 

  14. Gallego, M. A., Ballot, C., Kluza, J., Hajji, N., Martoriati, A., Castera, L., Cuevas, C., Formstecher, P., Joseph, B., Kroemer, G., Bailly, C., and Marchetti, P. (2007) Overcoming chemoresistance of non-small cell lung carcinoma through restoration of an AIF-dependent apoptotic pathway, Oncogene.

    Google Scholar 

  15. Jarve, A., Muller, J., Kim, I. H., Rohr, K., MacLean, C., Fricker, G., Massing, U., Eberle, F., Dalpke, A., Fischer, R., Trendelenburg, M. F., and Helm, M. (2007) Surveillance of siRNA integrity by FRET imaging, Nucleic Acids Res 35, e124.

    Google Scholar 

  16. Lin, Y. C., Hsieh, L. C., Kuo, M. W., Yu, J., Kuo, H. H., Lo, W. L., Lin, R. J., Yu, A. L., and Li, W. H. (2007) Human TRIM71 and Its Nematode Homologue Are Targets of let-7 MicroRNA and Its Zebrafish Orthologue Is Essential for Development, Mol Biol Evol 24, 2525–2534.

    Google Scholar 

  17. Khanam, T., Raabe, C. A., Kiefmann, M., Handel, S., Skryabin, B. V., and Brosius, J. (2007) Can ID Repetitive Elements Serve as Cis-acting Dendritic Targeting Elements? An In Vivo Study, PLoS ONE 2, e961.

    Google Scholar 

  18. Bossi, E., Fabbrini, M. S., and Ceriotti, A. (2007) Exogenous protein expression in Xenopus oocytes: basic procedures, Methods Mol Biol 375, 107–131.

    Google Scholar 

  19. Venoux, M., Basbous, J., Berthenet, C., Prigent, C., Fernandez, A., Lamb, N. J., and Rouquier, S. (2007) ASAP is a novel substrate of the oncogenic mitotic kinase Aurora-A : phosphorylation on Ser625 is essential to spindle formation and mitosis, Hum Mol Genet.

    Google Scholar 

  20. Jaulin, F., Xue, X., Rodriguez-Boulan, E., and Kreitzer, G. (2007) Polarization-dependent selective transport to the apical membrane by KIF5B in MDCK cells, Dev Cell 13, 511–522.

    Google Scholar 

  21. Derheimer, F. A., O’Hagan, H. M., Krueger, H. M., Hanasoge, S., Paulsen, M. T., and Ljungman, M. (2007) RPA and ATR link transcriptional stress to p53, Proc Natl Acad Sci USA 104, 12778–12783.

    Google Scholar 

  22. Bostrom, P., Andersson, L., Rutberg, M., Perman, J., Lidberg, U., Johansson, B. R., Fernandez-Rodriguez, J., Ericson, J., Nilsson, T., Boren, J., and Olofsson, S. O. (2007) SNARE proteins mediate fusion between cytosolic lipid droplets and are implicated in insulin sensitivity, Nat Cell Biol 9, 1286–1293.

    Google Scholar 

  23. Broders-Bondon, F., Chesneau, A., Romero-Oliva, F., Mazabraud, A., Mayor, R., and Thiery, J. P. (2007) Regulation of XSnail2 expression by Rho GTPases, Dev Dyn 236, 2555–2566.

    Google Scholar 

  24. Woodward, B. J., Campbell, K. H., and Ramsewak, S. S. (2007) A comparison of headfirst and tailfirst microinjection of sperm at intracytoplasmic sperm injection, Fertil Steril.

    Google Scholar 

  25. Yoshida, N., and Perry, A. C. (2007) Piezo-actuated mouse intracytoplasmic sperm injection (ICSI), Nat Protoc 2, 296–304.

    Google Scholar 

  26. Ehrenberg, M., and McGrath, J. L. (2005) Binding between particles and proteins in extracts: implications for microrheology and toxicity, Acta Biomater 1, 305–315.

    Google Scholar 

  27. Terns, M. P., and Goldfarb, D. S. (1998) Nuclear transport of RNAs in microinjected Xenopus Oocytes., Method Cell Biol 53, 559–589.

    Google Scholar 

  28. Fish, M. P., and Groth, A. C. (2007) Creating transgenic Drosophila by microinjecting the site-specific phiC31 integrase mRNA and a transgene-containing donor plasmid., Nat Protoc 2, 2325–2331.

    Google Scholar 

  29. Anand, R. J., Dai, S., Rippel, C., Leaphart, C., Qureshi, F., Gribar, S. C., Kohler, J. W., Li, J., Beer Stolz, D., Sodhi, C., and Hackam, D. J. (2008) Activated Macrophages Inhibit Enterocyte Gap Junctions via the Release of Nitric Oxide, Am J Physiol Gastrointest Liver Physiol 294, G109–119.

    Google Scholar 

  30. Green, M., Thorburn, A., Kern, R., and Loewenstein, P. M. (2007) The use of cell microinjection for the in vivo analysis of viral transcriptional regulatory protein domains, Methods Mol Med 131, 157–186.

    Google Scholar 

  31. Giovannardi, S., Soragna, A., Magagnin, S., and Faravelli, L. (2007) Functional expression of type 1 rat GABA transporter in microinjected Xenopus laevis oocytes, Methods Mol Biol 375, 235–255.

    Google Scholar 

  32. Hou, J. F., Cui, J., Yu, L. C., and Zhang, Y. (2009) Intracellular amyloid induces impairments on electrophysiological properties of cultured human neurons, Neurosci Lett 462, 294–299.

    Google Scholar 

  33. Bounhar, Y., Zhang, Y., Goodyer, C. G., and LeBlanc, A. (2001) Prion protein protects human neurons against Bax-mediated apoptosis, J Biol Chem 276, 39145–39149.

    Google Scholar 

  34. Guo, H., Albrecht, S., Bourdeau, M., Petzke, T., Bergeron, C., and LeBlanc, A. C. (2004) Active caspase-6 and caspase-6-cleaved tau in neuropil threads, neuritic plaques, and neurofibrillary tangles of Alzheimer’s disease, Am J Pathol 165, 523–531.

    Google Scholar 

  35. Guo, H., Petrin, D., Zhang, Y., Bergeron, C., Goodyer, C. G., and LeBlanc, A. C. (2006) Caspase-1 activation of caspase-6 in human apoptotic neurons, Cell Death Differ 13, 285–292.

    Google Scholar 

  36. Roucou, X., Giannopoulos, P. N., Zhang, Y., Jodoin, J., Goodyer, C. G., and LeBlanc, A. (2005) Cellular prion protein inhibits proapoptotic Bax conformational change in human neurons and in breast carcinoma MCF-7 cells, Cell Death Differ 12, 783–795.

    Google Scholar 

  37. Roucou, X., Guo, Q., Zhang, Y., Goodyer, C. G., and LeBlanc, A. C. (2003) Cytosolic prion protein is not toxic and protects against Bax-mediated cell death in human primary neurons, J Biol Chem 278, 40877–40881.

    Google Scholar 

  38. Tounekti, O., Zhang, Y., Klaiman, G., Goodyer, C. G., and LeBlanc, A. (2004) Proteasomal degradation of caspase-6 in 17beta-estradiol-treated neurons, J Neurochem 89, 561–568.

    Google Scholar 

Download references

Acknowledgments

This work was supported by the National Program of Basic Research sponsored by the Ministry of Science and Technology of China (2009CB941301), Peking University President Research Grant, Ministry of Education Recruiting Research Grant, and Roche Research Grant.

The authors declare no competing of interest. All authors declare no actual or potential conflicts of interest including any financial, personal, or other relationships with other people or organizations within 3 years of beginning the work submitted that could inappropriately influence (bias) their work.

Author information

Authors and Affiliations

  1. Laboratory of Neurobiology and State Key Laboratory of Biomembrane and Membrane Biotechnology, School of Life Sciences, Peking University, Beijing, China

    Yan Zhang

Authors
  1. Yan Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Yan Zhang .

Editor information

Editors and Affiliations

  1. POOLIA Sweden, Warfvinges väg 20, Stockholm, 104 25, Sweden

    Sara Lindström

  2. , Division of Nanobiotechnology, Royal Institute of Technology, Roslagstullsbacken 21, Stockholm, 106 91, Sweden

    Helene Andersson-Svahn

Rights and permissions

Reprints and permissions

Copyright information

© 2012 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Zhang, Y. (2012). Single-Cell Microinjection Technologies. In: Lindström, S., Andersson-Svahn, H. (eds) Single-Cell Analysis. Methods in Molecular Biology, vol 853. Humana Press. https://doi.org/10.1007/978-1-61779-567-1_13

Download citation

  • DOI: https://doi.org/10.1007/978-1-61779-567-1_13

  • Published:

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-566-4

  • Online ISBN: 978-1-61779-567-1

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation