Biology Reference
In-Depth Information
CHAPTER
7
Studying Mitochondria and
Microtubule Localization and
Dynamics in Standardized
Cell Shapes
Andrea Pelikan
*
, James Sillibourne
*
, Stephanie Miserey-Lenkei
*
, Frederique
Carlier-Grynkorn
*
, Bruno Goud
*
and Phong T. Tran
*
,{
*
Cell Biology, Institut Curie, UMR 144 CNRS, Paris, France
{
Cell and Developmental Biology, University of Pennsylvania, Philadelphia, Pennsylvania, USA
CHAPTER OUTLINE
Introduction .............................................................................................................. 98
7.1 Design of the Photomask .................................................................................... 99
7.2 Manufacturing of Micropatterned Coverslips...................................................... 100
7.3 Seeding and Spreading of Cells ........................................................................ 101
7.4 Mitochondria and Microtubule Visualization ...................................................... 102
7.4.1 Mitochondria and Microtubule Labeling............................................ 102
7.4.2 Live-cell Visualization ..................................................................... 103
7.5 Materials......................................................................................................... 104
7.5.1 Preparation of Micropatterns ........................................................... 104
7.5.2 Seeding and Spreading of Cell ......................................................... 105
7.5.3 Mitochondria and Microtubule Visualization...................................... 105
Conclusion ............................................................................................................. 106
Acknowledgments ................................................................................................... 107
References ............................................................................................................. 107
Abstract
Mammalian cells show a large diversity in shape and are both shape-changing and
mobile when cultured on conventional uniform substrates. The use of micropattern-
ing techniques limits the number of variable parameters, by imposing shape and stan-
dardized adhesive areas on the cells, which facilitates analysis. By changing size or
