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Isolation of bacteria-containing phagosomes by magnetic selection.

Lönnbro, Per LU ; Nordenfelt, Pontus LU and Tapper, Hans LU (2008) In BMC Cell Biology 9(June 27).
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... (More)
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. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
BMC Cell Biology
volume
9
issue
June 27
publisher
BioMed Central
external identifiers
  • WOS:000257835800001
  • PMID:18588680
  • Scopus:47749127370
ISSN
1471-2121
DOI
10.1186/1471-2121-9-35
language
English
LU publication?
yes
id
01b85614-3a33-432c-a544-912bb45b8b52 (old id 1181753)
alternative location
http://www.ncbi.nlm.nih.gov/pubmed/18588680?dopt=Abstract
date added to LUP
2008-08-06 10:49:02
date last changed
2016-12-04 04:39:23
@misc{01b85614-3a33-432c-a544-912bb45b8b52,
  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.},
  author       = {Lönnbro, Per and Nordenfelt, Pontus and Tapper, Hans},
  issn         = {1471-2121},
  language     = {eng},
  number       = {June 27},
  publisher    = {ARRAY(0xb78bd20)},
  series       = {BMC Cell Biology},
  title        = {Isolation of bacteria-containing phagosomes by magnetic selection.},
  url          = {http://dx.doi.org/10.1186/1471-2121-9-35},
  volume       = {9},
  year         = {2008},
}