B. Murienne - Master Project Thesis - Infoscience - EPFL

Contents
Contents......................................................................................................................................2
Abstract ......................................................................................................................................3
1. Introduction.............................................................................................................................4
2. Theory ....................................................................................................................................6
2.1 The heart...........................................................................................................................6
2.2 Photolithography.............................................................................................................11
2.3 Stretch devices ................................................................................................................12
2.4 Electrodes .......................................................................................................................13
2.5 Optical mapping..............................................................................................................14
2.6 Post-processing of the optical signals...............................................................................17
2.7 Immunostaining ..............................................................................................................21
3. Materials and methods...........................................................................................................22
3.1 Cardiomyocyte isolation..................................................................................................22
3.2 Photolithography.............................................................................................................22
3.3 Fabrication of micropatterned elastic silicone membranes................................................23
3.4 Cardiomyocyte culture on micropatterned elastic silicone membranes .............................24
3.5 Staining procedure design for optical mapping of cardiomyocyte monolayers..................25
3.6 Electrode design and cardiomyocyte monolayer pacing ...................................................26
3.7 Immunostaining ..............................................................................................................27
3.8 Temperature and oxygenation setup design......................................................................27
3.9 Stretcher calibration ........................................................................................................29
3.10 Optical setup for cardiomyocyte monolayer imaging .....................................................29
3.11 Acquisition setup...........................................................................................................30
3.12 Extraction of activation time, repolarization, APD and CV values..................................31
4. Results ..................................................................................................................................32
4.1 Cardiomyocyte culture on silicone micropatterned membranes........................................32
4.2 Staining and optical signal recording ...............................................................................33
4.3 Making new micropatterned silicon wafers......................................................................34
4.4 Cardiomyocyte pacing.....................................................................................................36
4.5 Analysis ..........................................................................................................................38
4.6 Immunostaining ..............................................................................................................39
4.7 Design of a temperature control system ...........................................................................41
4.8 Stretcher calibration ........................................................................................................42
4.9 Optical mapping experiments ..........................................................................................42
5. Discussion.............................................................................................................................45
6. Conclusion ............................................................................................................................49
References ................................................................................................................................50
Acknowledgments.....................................................................................................................53
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