Abstract
The ability to microinject substances into the cytosol of living neutrophils opens the possibility of manipulating the chemistry within the cell and also of monitoring changes using indicators which otherwise cannot be introduced into the cell. However, neutrophils cannot be microinjected by the conventional glass pipette insertion method. Here, we outline two techniques which work well with neutrophils, namely, SLAM (simple lipid-assisted microinjection) and electroinjection.
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References
Guse AH, Berg I, da Silva CP et al (1997) Ca2+ entry by cyclic ADP-ribose in intact T-lymphocytes. J Biol Chem 272:8546–8550
Hallett MB, Campbell AK (1980) Uptake of liposomes containing the photoprotein obelin by rat isolated adipocytes; adhesion, endocytosis or fusion? Biochem J 192:587–596
Gao X, Huang L (1995) Cationic liposome-mediated gene transfer. Gene Ther 2:710–722
Hallett MB, Campbell AK (1982) Measurement of changes in cytoplasmic free calcium in fused cell hybrids. Nature 294:155–158
Campbell AK, Hallett MB (1983) Measurement of intracellular calcium ions and oxygen radicals in polymorphonuclear leukocyte–erythrocyte “ghost” hybrids. J Physiol 338:537–550
Laffafian I, Hallett MB (1998) Lipid-assisted microinjection: introducing material into the cytosol and membranes of small cells. Biophys J 75:2558–2563
Peters R, Sikorski R (1998) Gentle slam. Science 282:2213–2214
Laffafian I, Hallett MB (2000) Gentle micro-injection for myeloid cells using SLAM. Blood 95:3270–3271
Davies-Cox EV, Laffafian I, Hallett MB (2001) Control of Ca2+ influx in human neutrophils by IP3 binding: differential effects of micro-injected IP3 receptor antagonists. Biochem J 355:139–143
Dewitt S, Laffafian I, Hallett MB (2003) Phagosomal oxidative activity during β2 integrin (CR3)-mediated phagocytosis by neutrophils is triggered by a non-restricted Ca2+ signal: Ca2+ controls time not space. J Cell Sci 116:2857–2865
Zimmermann U (1986) Electrical breakdown, electropermeabilization and electrofusion. Rev Physiol Biochem Pharmacol 105:175–256
Haas K, Sin WC, Javaherian A et al (2001) Single-cell electroporation for gene transfer in vivo. Neuron 29:583–591
Haas K, Jensen K, Sin WC et al (2002) Targeted electroporation in Xenopus tadpoles in vivo – from single cells to the entire brain. Differentiation 70:148–154
Bestman JE, Ewald RC, Chiu SL, Cline HT (2006) In vivo single-cell electroporation for transfer of DNA and macromolecules. Nat Protoc 1:1267–1272
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Laffafian, I., Lewis, K.J., Masterman, K.B., Hallett, M.B. (2014). Microinjection Methods for Neutrophils. In: Quinn, M., DeLeo, F. (eds) Neutrophil Methods and Protocols. Methods in Molecular Biology, vol 1124. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-845-4_11
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DOI: https://doi.org/10.1007/978-1-62703-845-4_11
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-844-7
Online ISBN: 978-1-62703-845-4
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