We study the action of the group G = GL_2(C) of invertible matrices over the complex numbers on the complex vector space V of n-tuples of 2x2 matrices. The algebra of invariants C[V]^G for this action is well-known, and has dimension 4n-3 and minimum generating set E_n with cardinality 1/6(n^3+11n). In recent work, Kaygorodov, Lopatin and Popov showed that this generating set is also a minimal separating set by inclusion, i.e. no proper subset is a separating set. This does not mean it has smallest possible cardinality among all separating sets. We show that if S is a separating set for C[V]^G then |S| is at least 5n-5. In particular for n=3, the set E_n is indeed of minimal cardinality, but for n>3 may not be so. We then show that a smalle...