2. Modelling Cell Attachment Jonathan A. Ratcliffe

3. Outline Aims Theory Imaging Image Processing Comparison to SEM Conclusions Future work

4. Aims To visualise a cell as it undergoes the transition from attaching and spreading to crawling To create an accurate three-dimensional representation of the cell so that predictions can be made about spreading using the dynamic contact line of the cell.

5. The dynamic contact angle

6. 2 stages of cell spreading: Passive and Active

7. Theory J. Oliver, et al. Thin-film theories for two-phase reactive flow models of active cell motion Mathematical Medicine and Biology (2005) 22, 53–98 Dynamic Contact angle (to be determined) Static contact angle Network density at contact line Mass transfer rate from G- to F- actin at contact line Outward normal velocity for contact line. Fluid forcesMotile machinery

8. Imaging LavaCell™ stain chosen as: Cost effective Simple to use, add 1.8μl for 1.5ml media Long stokes shift (fluorescence lifetime) No cytotoxicity observed up to 24h. Best images

10. Image Processing

11. Image Prossesing

15. Cell imaged at 20 minutes after seeding.

16. Cell imaged 60 minutes after seeding.

17. Human Osteosarcoma cell 70 minutes after seeding.

18. Conclusions Image processing technique accurately finds outline of cell segments. 3D representations visually similar to SEM images of fixed cells.

19. Future Work To look at transparent coated surfaces such as: TiN, TiO 2, Gold, SS 316L. Patterned surfaces Surface topography using embossed glass