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BODIPY-ceramide labeling and live imaging of Chlamydia

JC John V. Cox
YA Yasser Mohamed Abdelrahman
SO Scot P. Ouellette
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HeLa cells were infected with C. trachomatis serovar L2. At 9.5 hpi 5 µM BODIPY FL C5 ceramide complexed with BSA was added to the media and the infected cells were incubated at 4 °C for 30 min5,59. The cells were then washed twice with DMEM and incubated with DMEM/0.7% BSA for one hour to "back-exchange" BODIPY FL C5 ceramide from the plasma membrane. Cells were imaged using a Zeiss LSM710 confocal microscope equipped with a 63 × Plan-Apochromat oil immersion lens. Alternatively, cells were imaged using a 63 × Plan-Apochromat immersible lens on a Zeiss AxioImager. M2 microscope equipped with heatable Universal Mounting Frame and images were collected using an AxioCam MRm camera.

Copyright and License information: The Author(s) ©2020
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