Description

Disruption of Microtubules with Nocodazol U2OS human osteosarcoma cells were transduced with 'Cellular Lights' Actin-RFP (Actin-red fluorescent protein) and Cellular Lights MAP4-GFP (MAP4-green fluorescent protein) and imaged the next day. Cells were washed in HBSS and incubated with 1 ug/ml of the DNA stain Hoechst 33342 for 10 minutes then washed with HBSS. Nocodazole, a microtubule inhibitor, was applied to the bath at a final concentration of 10 uM. Images were taken every 20 seconds for 30 minutes. In this video, microtubules appear green, actin microfilaments appear red, and the nucleus is stained blue. Note the loss of the microtubular network (green) whilst the actin cytoskeleton (red) remains intact. Cells were imaged on a Deltavision core microscope with a 40X objective, 1.4 NA. Fluorescence was detected using the appropriate filters (DAPI/FITC/TRITC). The time series was deconvolved using standard software. Scale bar = 20 microns.