Front Cover; Methods in Cell Biology; Series Editors; Methods in Cell BiologyCell Polarity and MorphogenesisVolume 139Edited byThomas LecuitIBDM (Institut de Biologie du Dévelop ... ; Contents; Contributors; 1 -- A user's guide for characterizing plasma membrane subdomains in living cells by spot variation fluorescence cor ... ; INTRODUCTION; 1. BUILDING A SENSITIVE AND ROBUST HOMEMADE SPOT VARIATION FLUORESCENCE CORRELATION SPECTROSCOPY/FLUORESCENCE RECOVERY AFTER P ... ; 1.1 Description of the Optical Bench and Signal Collection; 1.2 Common Pitfalls and Practical Considerations

2. SAMPLE PREPARATIONS AND LABELING2.1 General Considerations on Sample Preparations; 2.2 Use of Fluorescent Proteins; 2.3 Membrane Labeling With Lipid Probes. Impact on Fluorescence Correlation Spectroscopy Measurements; 2.4 Surface Labeling of Nonadherent Cells With Antibodies Prior to Fluorescence Correlation Spectroscopy Measurements; 2.5 Pharmacological Treatments; 2.5.1 Latrunculin A treatment (actin meshwork disassembly); 2.5.2 Sphingolipid and cholesterol metabolism inhibition; 3. FLUORESCENCE CORRELATION SPECTROSCOPY RECORDING; 3.1 Preliminary Recording; 3.2 Spot Size Calibration

3.3 Fluorescence Correlation Spectroscopy Acquisitions at the Plasma Membrane of Living Cells3.4 Spot Fluorescence Recovery After Photobleaching Measurements at the Plasma Membrane of Living Cells; 4. DATA ANALYSIS AND INTERPRETATION; 4.1 Data Analysis and Fitting Procedures; 4.2 Data Interpretation; 4.3 Comparison of Spot Variation Fluorescence Correlation Spectroscopy Results; CONCLUSION; Acknowledgments; REFERENCES; 2 -- Investigating symmetry breaking in yeast: from seeing to understanding; INTRODUCTION; 1. GENETIC TOOLS; 2. MICROSCOPY-BASED METHODS TO VISUALIZE CELL POLARIZATION

2.1 Single Cell Time-Lapse Imaging2.1.1 Slide preparation for short-time lapse imaging; 2.1.2 Slide preparation for long time-lapse imaging; 2.2 Bulk Cell Polarization Assays; 2.3 Fluorescence Recovery After Photobleaching; 2.3.1 Practical considerations for fluorescence recovery after photobleaching in Saccharomyces cerevisiae; 2.3.2 Mathematical modeling of the fluorescence recovery after photobleaching data; 2.4 Fluorescence Correlation Spectroscopy; 2.4.1 Requirements for fluorescence correlation spectroscopy; 2.4.2 Fluorescence cross-correlation spectroscopy

2.4.3 Requirements and analysis of fluorescence cross-correlation spectroscopy2.5 Fluorescence Resonance Energy Transfer; 2.5.1 Design of fluorescence resonance energy transfer biosensor for the guanine nucleotide exchange factor activity toward Cdc42; 2.5.2 Fluorescence resonance energy transfer experiments with the Cdc42 activation biosensor; CONCLUSION AND FUTURE PERSPECTIVE; NOTES; Acknowledgments; REFERENCES; 3. Precise tracking of the dynamics of multiple proteins in endocytic events; INTRODUCTION; 1. RATIONALE; 2. SAMPLE PREPARATION; 3. FLUORESCENCE MICROSCOPY; 3.1 Microscopy Setup