TY - JOUR
T1 - Border cell migration
T2 - A model system for live imaging and genetic analysis of collective cell movement
AU - Prasad, Mohit
AU - Wang, Xiaobo
AU - He, Li
AU - Cai, Danfeng
AU - Montell, Denise J.
N1 - Publisher Copyright:
© 2015, Springer Science+Business Media New York.
PY - 2015
Y1 - 2015
N2 - Border cell migration in the Drosophila ovary has emerged as a genetically tractable model for studying collective cell movement. Over many years border cell migration was exclusively studied in fixed samples due to the inability to culture stage 9 egg chambers in vitro. Although culturing late-stage egg chambers was long feasible, stage 9 egg chambers survived only briefl y outside the female body. We identifi ed culture conditions that support stage 9 egg chamber development and sustain complete migration of border cells ex vivo. This protocol enables one to compare the dynamics of egg chamber development in wild-type and mutant egg chambers using time-lapse microscopy and taking advantage of a multiposition microscope with a motorized imaging stage. In addition, this protocol has been successfully used in combination with fluorescence resonance energy transfer biosensors, photo-activatable proteins, and pharmacological agents and can be used with wide-field or confocal microscopes in either an upright or an inverted configuration.
AB - Border cell migration in the Drosophila ovary has emerged as a genetically tractable model for studying collective cell movement. Over many years border cell migration was exclusively studied in fixed samples due to the inability to culture stage 9 egg chambers in vitro. Although culturing late-stage egg chambers was long feasible, stage 9 egg chambers survived only briefl y outside the female body. We identifi ed culture conditions that support stage 9 egg chamber development and sustain complete migration of border cells ex vivo. This protocol enables one to compare the dynamics of egg chamber development in wild-type and mutant egg chambers using time-lapse microscopy and taking advantage of a multiposition microscope with a motorized imaging stage. In addition, this protocol has been successfully used in combination with fluorescence resonance energy transfer biosensors, photo-activatable proteins, and pharmacological agents and can be used with wide-field or confocal microscopes in either an upright or an inverted configuration.
KW - Border cell migration
KW - Collective cell migration
KW - Drosophila stage 9 egg chambers
KW - Organ culture
KW - Time-lapse live imaging
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U2 - 10.1007/978-1-4939-2851-4_6
DO - 10.1007/978-1-4939-2851-4_6
M3 - Article
C2 - 26324431
AN - SCOPUS:84940832804
SN - 1064-3745
VL - 1328
SP - 89
EP - 97
JO - Methods in Molecular Biology
JF - Methods in Molecular Biology
ER -