An approach to quantitatively study vesicle dynamics as well as biologically-related micro-objects in a fluid flow, which is based on the combination of a dynamical trap and a control parameter, the ratio of the vorticity to the strain rate, is suggested. The flow is continuously varied between rotational, shearing, and elongational in a microfluidic 4-roll mill device, the dynamical trap, that allows scanning of the entire phase diagram of motions, i.e., tank-treading (TT), tumbling (TU), and trembling (TR), using a single vesicle even at ؍ in/out ؍ 1, where in and out are the viscosities of the inner and outer fluids. This cannot be achieved in pure shear flow, where the transition between TT and either TU or TR is attained only at >1. As a result, it is found that the vesicle dynamical states in a general are presented by the phase diagram in a space of only 2 dimensionless control parameters. The findings are in semiquantitative accord with the recent theory made for a quasi-spherical vesicle, although vesicles with large deviations from spherical shape were studied experimentally. The physics of TR is also uncovered. U nderstanding the rheology of biofluids remains a great challenge, whose progress relies, in a large part, on detailed studies of the dynamics of a single cell. Vesicles are a model system used to study the dynamic behavior of biological cells, similar in some respects to red blood cells, and their dynamics in shear flow have been the subject of intensive theoretical (1-8), numerical (9-13), and experimental (14-18) investigations.A vesicle is a droplet of viscous fluid encapsulated by a phospholipid bilayer membrane suspended in a fluid of either the same or different viscosity as the inner one. Both the volume and the surface area of the vesicle are conserved. The former means that the vesicle membrane is considered to be impermeable, at least on the time scale of the experiment, and the latter means that the membrane dilatation is neglected since it is 2D fluid (1,2). Experimental, theoretical, and computational efforts during the last decade led to the observation and characterization of 3 states in vesicle dynamics in shear flow. The existence of the first 2, tank-treading (TT) and tumbling (TU), and the transition between them were already predicted by a phenomenological model of Keller and Skalak (19) and its further extensions (2,11,12). Two control parameters, the excess area ⌬ ϭ A/R 2 -4 and the viscosity contrast ϭ in / out , determine the transition line c (⌬) between TT and TU, which is independent of the shear rate ␥ in the approximation of a fixed vesicle shape, with the vesicle inclination angle with respect to the flow direction as the only dynamical variable (2,9,10,16-19). Here, R is the effective vesicle radius, related to the volume via V ϭ 4/3R 3 , A is the vesicle surface area, and in and out are the viscosities of the inner and outer fluids. Another analytical approach based on a quasi-spherical vesicle approximated by a spherical harmonics expansion used a perturb...