Lab tek 8 well imaging chambers

Beas2B cells expressing mTagBFP2::EML4-ALK and GFP::DCP1B were seeded into Nunc Lab-Tek 8-well imaging chambers. A custom-made sample holder ensured the imaging chamber fits securely on the microscope stage without position shift. The cells were first imaged in regular RPMI cell culture media and then the media was replaced by RPMI media containing 5% hexanediol (Sigma). The same field of view was monitored at defined time points after addition of hexanediol.

Beas2B cells were seeded into Nunc Lab-Tek 8-well imaging chambers and plasmids encoding wild-type and mutant forms of mTagBFP2::EML4-ALK (i.e., ΔTD, ΔHELP, K589M, see text for details), mTagBFP2::CCDC6-RET (i.e., ΔCC, K147M), as well as YFP::EML4-ALK variants 1, 3, and 5 were transfected using Mirus TransIT-LT1. 24 hours later, an initial position in each well was randomly picked as the center of an area of 650 μm x 650 μm and imaged with automated scanning and tiling done through MicroManager with a 100x oil objective (N.A. = 1.40). The process was repeated three times and all the cells were scored manually to determine if they contained cytoplasmic granules.