U2OS-Cell Cycle Chromobody®-TagRFP
U2OS cell line stably expressing Cell Cycle Chromobody® fused to the red fluorescent protein TagRFP
VHH antibody fragment against cell cycle marker PCNA (proliferating cell nuclear antigen)Actin-Chromobody®
Live cell imaging of U2OS-Cell Cycle Chromobody®-TagRFP
For live cell imaging cells were seeded in ibidi µ-Slides. Time series was acquired on a spinning disk microscope (UltraVIEW VoX, PerkinElmer) equipped with a 63x/1.4 NA Plan-Apochromat oil immersion objective and a humidified, temperature- and CO2-controlled live cell chamber. TagRFP was excited with a 561 nm laser-line for typically between 100-500 ms/ image with laser intensity set to max. 10%. Confocal image z-stacks of living cells were recorded with a frame size of 1000x1000 pixels, a pixel size of 100 nm and a z-step size of 400 nm every 30 minutes. Images shown are maximum intensity projections of few mid-z-sections.