For Hopf thresholds in a two-pool calcium model, with the unique equilibrium parametrized by , locate changes of stability by solving the two equations
If there are two simple roots with positive trace between them, the equilibrium is a repeller throughout that interval, since its determinant stays positive. At a nondegenerate crossing a Hopf bifurcation is possible, with linear frequency . A complete classification of criticality requires a nonlinear coefficient, not merely the trace calculation. At leading singular order the thresholds approach the folds determined by and , equivalently .
Periodic solutions are also supported globally in the positive quadrant. The axes point inward for positive input, bounds , and the equation then bounds by a linear restoring term plus a bounded source. A large rectangle can thus be chosen forward invariant. For an initial point distinct from the repelling unique equilibrium, its omega-limit set cannot contain that equilibrium or a boundary equilibrium. The Poincaré-Bendixson theorem supplies a periodic orbit. In the strong separation regime it follows the two attracting branches and jumps between them, a relaxation oscillation.
The interval requires the maximum of along the critical manifold to exceed ; the listed scale hierarchy alone is not a substitute for that test for arbitrary remaining constants. As an explicit example, , , and give approximately and . At the trace is positive, whereas at and it is negative. The figures use this example and label their singular-limit portions accordingly.

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