Monitoring mitotic cell division in DT40 cells - Events

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Setting up DT40s for time-lapse microscopy (I) Prepare coverslips • Coat 12 mm round coverslips with poly-L-lysine, 10 μg/ ml in sterile PBS, O/N, at 4 o C • Wash a couple of times with PBS: coverslips are ready to use. You can keep coated coverslips in PBS at 4 o C for a few days • If in a hurry, you can also coat coverslips at 37 o C for 2 hrs and then wash in PBS and use
Set up cells Setting up DT40s for time-lapse microscopy (II) • On the previous day, count cells expressing H2B:GFP, spin them down and resuspend in fresh medium at approximately 1 x10 5 cells/ ml • Place a poly-L-lysine-coated coverslip inside an empty 30 mm petri dish • Drop 100 μl (i.e. 1 x 10 4 ) cells onto the coverslip (you don’t want cells too crowded!!) • Leave at room temperature for approximately 1h for DT40s to sit down • Carefully put 2 ml growth medium inside the 30 mm dish and return to the incubator • Next day, monitor cells on the time-lapse microscope and get still photos or movies!!
Page 1 and 2: Tubulin DNA Monitoring mitotic cell
Page 3 and 4: Time-lapse video microscopy • Cel
Page 5 and 6: Components of a time-lapse video mi
Page 7 and 8: Components of a time-lapse video mi
Page 9 and 10: Generation of DT40 cells stably exp
Page 11 and 12: Electroporation of DT40 cells (I) I
Page 13: Isolation of DT40 clones • The ne
Page 17 and 18: Monitoring mitotic cell division by
Page 19 and 20: metaphase anaphase Normal chromosom
Page 21 and 22: Chk1-deficient Chk1 deficient DT40
Page 23: Living cell Apoptotic cell Mitotic

Monitoring mitotic cell division in DT40 cells - Events

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