Modelling Cell Attachment Jonathan A. Ratcliffe Outline Aims Theory Imaging Image Processing...

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Modelling Cell Attachment Jonathan A. Ratcliffe
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Transcript of Modelling Cell Attachment Jonathan A. Ratcliffe Outline Aims Theory Imaging Image Processing...

Page 1: Modelling Cell Attachment Jonathan A. Ratcliffe Outline Aims Theory Imaging Image Processing Comparison to SEM Conclusions Future work.

Modelling Cell AttachmentJonathan A. Ratcliffe

Page 2: Modelling Cell Attachment Jonathan A. Ratcliffe Outline Aims Theory Imaging Image Processing Comparison to SEM Conclusions Future work.

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

Page 3: Modelling Cell Attachment Jonathan 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.

Page 4: Modelling Cell Attachment Jonathan A. Ratcliffe Outline Aims Theory Imaging Image Processing Comparison to SEM Conclusions Future work.

The dynamic contact angle

Page 5: Modelling Cell Attachment Jonathan A. Ratcliffe Outline Aims Theory Imaging Image Processing Comparison to SEM Conclusions Future work.

2 stages of cell spreading: Passive and Active

Page 6: Modelling Cell Attachment Jonathan A. Ratcliffe Outline Aims Theory Imaging Image Processing Comparison to SEM Conclusions Future work.

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

Page 7: Modelling Cell Attachment Jonathan A. Ratcliffe Outline Aims Theory Imaging Image Processing Comparison to SEM Conclusions Future work.

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

Page 8: Modelling Cell Attachment Jonathan A. Ratcliffe Outline Aims Theory Imaging Image Processing Comparison to SEM Conclusions Future work.
Page 9: Modelling Cell Attachment Jonathan A. Ratcliffe Outline Aims Theory Imaging Image Processing Comparison to SEM Conclusions Future work.

Image Processing

Page 10: Modelling Cell Attachment Jonathan A. Ratcliffe Outline Aims Theory Imaging Image Processing Comparison to SEM Conclusions Future work.

Image Prossesing

Page 11: Modelling Cell Attachment Jonathan A. Ratcliffe Outline Aims Theory Imaging Image Processing Comparison to SEM Conclusions Future work.
Page 12: Modelling Cell Attachment Jonathan A. Ratcliffe Outline Aims Theory Imaging Image Processing Comparison to SEM Conclusions Future work.
Page 13: Modelling Cell Attachment Jonathan A. Ratcliffe Outline Aims Theory Imaging Image Processing Comparison to SEM Conclusions Future work.
Page 14: Modelling Cell Attachment Jonathan A. Ratcliffe Outline Aims Theory Imaging Image Processing Comparison to SEM Conclusions Future work.

Cell imaged at 20 minutes after seeding.

Page 15: Modelling Cell Attachment Jonathan A. Ratcliffe Outline Aims Theory Imaging Image Processing Comparison to SEM Conclusions Future work.

Cell imaged 60 minutes after seeding.

Page 16: Modelling Cell Attachment Jonathan A. Ratcliffe Outline Aims Theory Imaging Image Processing Comparison to SEM Conclusions Future work.

Human Osteosarcoma cell 70 minutes after seeding.

Page 17: Modelling Cell Attachment Jonathan A. Ratcliffe Outline Aims Theory Imaging Image Processing Comparison to SEM Conclusions Future work.

Conclusions

Image processing technique accurately finds outline of cell segments.

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

Page 18: Modelling Cell Attachment Jonathan A. Ratcliffe Outline Aims Theory Imaging Image Processing Comparison to SEM Conclusions Future work.

Future Work To look at transparent coated surfaces

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