modelling cell attachment jonathan a. ratcliffe outline aims theory imaging image processing...

Modelling Cell AttachmentJonathan A. Ratcliffe

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

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.

The dynamic contact angle

2 stages of cell spreading: Passive and Active

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 forces Motile machinery

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

Image Processing

Image Prossesing

Cell imaged at 20 minutes after seeding.

Cell imaged 60 minutes after seeding.

Human Osteosarcoma cell 70 minutes after seeding.

Conclusions

Image processing technique accurately finds outline of cell segments.

3D representations visually similar to SEM images of fixed cells.

Future Work To look at transparent coated surfaces

such as: TiN, TiO2, Gold, SS 316L. Patterned surfaces Surface topography using embossed glass