Osmotic up/down shock

1. Osmotic up/down shock A summary of Allison’s data 9/3/10

2. Upshift of Br strain Histograms of width and length for individual time points to follow with information about average width and number of cells Take home: 1 hour not enough, 2 hours enough.

3. Br Ala-Pro steady-state 570 cells with average width 0.71 microns

4. Upshift for 1,2,3,4 hours Looks like shift takes place between 1hr and 2hrs

5. Br upshift 1 hour 447 cells with average width 0.7 microns

6. Br upshift 2 hour 622 cells with average width 0.85 microns (fixed this run tonight 9/8/10)

7. Br upshift 3 hour 170 cells with average width 0.89 microns

8. Br upshift 4 hour 335 cells with average width 0.9 microns

9. Downshift of Br strain Histograms of width and length for individual time points to follow Take home: 4 hours may or may not be enough.

10. Br Glu-Caa steady-state 359 cells with average width 1.05 microns

11. Downshift for 2,4 hours Looks like shift still taking place after 4 hours

12. Br downshift 2 hour 1687 cells with average width 0.94 microns

13. Br downshift 4 hour 1247 cells with average width 0.86 microns

14. Upshift of MG1655 strain Histograms of width and length for individual time points to follow Take home: Looks like only small increase in width after 2 hours (slower adjustment than in Br strain). I’m going to check the analysis to make sure.

15. MG1655 Ala-Pro steady-state 323 cells with average width 0.68 microns

16. Upshift for 1,2 hours Looks like shifting very slowly

17. MG1655 upshift 1 hour 605 cells with average width 0.71 microns

18. MG1655 upshift 2 hour 779 cells with average width 0.7 microns

19. Downshift of MG1655 strain Histograms of width and length for individual time points to follow Take home: Looks same as Br strain. Needs at least 4 hours.

20. MG1655 Glu-Caa steady-state 445 cells with average width 0.97 microns

21. Downshift for 2,4 hours Same as Br, looks like shift is not complete after 4 hours

22. MG1655 downshift 2 hours 1298 cells with average width 0.8 microns

23. MG1655 downshift 4 hours 1106 cells with average width 0.74 microns

24. Summary • Colors are set to match up the same experiments between Br and MG1655 cells • Note the good agreement between the steady-state sizes of the two strains

25. Splitting between ceph-treated and non-treated cells… Take home message: The widths vary systematically, although the ceph-treated cells are systematically thinner. Incidentally, we’ve seen this in our own experiments…

26. Summary (ceph/non-ceph)

27. Growth curves for upshift

28. Growth curves for downshift

29. Br upshift 1 hour

30. Br upshift 2 hours

31. 3hr and 4hr time points I didn’t add these because there are so few cells that produce good data (they are so long that the edge parameterization doesn’t work very well).

32. MG1655 upshift 1 hour

33. MG1655 upshift 2 hours

34. MG1655 downshift 2 hours

35. MG1655 downshift 4 hours

36. Contour level • Note that cell width is sensitive to contour level of segmentation • The following slides show example galleries of images to illustrate how the cells are segmented. If you zoom in, you can see the cell outline. I’ve tried to vary the contour level to fit each frame as much as possible. • Note that it’s possible that the cells are in fact the same size, but that the contour levels are just different. I’m checking that out…. • Keep in mind that the cells are not drawn to scale. I’m trying to fix that but Matlab is retarded.

37. Br Ala-Pro steady-state

38. Br Ala-Pro steady-state

39. Br Ala-Pro steady-state

40. Br 2 hour upshift

41. Br 2 hour upshift

42. Br 2 hour upshift (zoom)

43. MG1655 2 hour upshift

44. MG1655 2 hour upshift

45. MG1655 2 hour upshift (zoom)