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Open Access Highly Accessed Methodology article

Isolation of bacteria-containing phagosomes by magnetic selection

Per Lönnbro, Pontus Nordenfelt and Hans Tapper*

Author Affiliations

Division of Infection Medicine, Department of Clinical Sciences, Lund University, SE-221 84 Lund, Sweden

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BMC Cell Biology 2008, 9:35  doi:10.1186/1471-2121-9-35

Published: 27 June 2008

Abstract

Background

There is a growing awareness of the importance of intracellular events in determining the outcome of infectious disease. To improve the understanding of such events, like phagosome maturation, we set out to develop a versatile technique for phagosome isolation that is rapid and widely applicable to different pathogens.

Results

We developed two different protocols to isolate phagosomes containing dead or live bacteria modified with small magnetic particles, in conjunction with a synchronized phagocytosis protocol and nitrogen cavitation. For dead bacteria, we performed analysis of the phagosome samples by microscopy and immunoblot, and demonstrated the appearance of maturation markers on isolated phagosomes.

Conclusion

We have presented detailed protocols for phagosome isolation, which can be adapted for use with different cell types and prey. The versatility and simplicity of the approach allow better control of phagosome isolation, the parameters of which are critical in studies of host-bacteria interaction and phagosome maturation.