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Fig. 6 | BMC Cell Biology

Fig. 6

From: An easy-to-build and re-usable microfluidic system for live-cell imaging

Fig. 6

Monitoring cellular responses to dynamic changes in their environment. a. Analogue-sensitive fission yeast cells [37] were injected in a lectin-coated chip (as in Additional file 4C) and perfused with 1 μM 3-MBPP1 (20 μL/min) at 32 °C for 2 h 45 min (G2 block) before being released in inhibitor-free medium (T = 0, 20 μL/min). Control: cells grown in batch cultures at 32 °C, treated with 1 μM 3-MBPP1 for 2 h 45 min and released into inhibitor-free medium by filtration (T = 0). The septation indexes were then monitored over time (n > 50 for each time point). Averages of 3 independent experiments are shown with standard errors. Note that images were acquired every 5 min for the chips (mounted on a microscope) but every 10 min for the flasks, for which 5 min intervals were difficult to achieve. This led to a higher variability in timing for the chip assays compared to the less resolved batch cultures. b. HeLa cells were injected in a microsystem (as in Fig. 4c) maintained at 37 °C on a high precision hot plate to facilitate this proof-of-concept assay. Cells were then allowed to adhere for 3 h and subsequently treated with 6 μM of the CDK inhibitor RO-3306 for 10 min (13 μL/min) every hour for a total of 20 h. Cells were subsequently released in inhibitor-free medium (T = 0; 15 min at 20 μL/min and then 5 μL/min), and the percentages of mitotic (rounded) and post-mitotic (doublets) cells were quantified over time. Measurements were made in an area within 1.5 mm on each side of the center of the channel. As a control, cells treated with 6 μM RO-3306 in dishes were washed and incubated in normal medium. Averages of 3 independent experiments (n > 100 for each experiment) are shown with standard errors

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