  Phase-contrast microscopy is used to monitor the shapes of micron-scale
fluid-phase phospholipid-bilayer vesicles in aqueous solution. At fixed
temperature, each vesicle undergoes thermal shape fluctuations. We are able
experimentally to characterize the thermal shape ensemble by digitizing the
vesicle outline in real time and storing the time-sequence of images. Analysis
of this ensemble using the area-difference-elasticity (ADE) model of vesicle
shapes allows us to associate (map) each time-sequence to a point in the
zero-temperature (shape) phase diagram. Changing the laboratory temperature
modifies the control parameters (area, volume, etc.) of each vesicle, so it
sweeps out a trajectory across the theoretical phase diagram. It is a
nontrivial test of the ADE model to check that these trajectories remain
confined to regions of the phase diagram where the corresponding shapes are
locally stable. In particular, we study the thermal trajectories of three
prolate vesicles which, upon heating, experienced a mechanical instability
leading to budding. We verify that the position of the observed instability and
the geometry of the budded shape are in reasonable accord with the theoretical
predictions. The inability of previous experiments to detect the ``hidden''
control parameters (relaxed area difference and spontaneous curvature) make
this the first direct quantitative confrontation between vesicle-shape theory
and experiment.
