Changes of dermal collagen and elastin content are characteristic for skin aging as well as for pathological skin conditions. To evaluate these changes, we used in vivo multiphoton laser tomography to measure two-photon excited autofluorescence (AF) and second harmonic generation (SHG). We tested 18 patients of all ages and calculated the SHG-to-AF aging index of dermis (SAAID). We observed a negative relationship between the SAAID and age, which was accelerated for the female (n=7) subgroup. The current findings are the first in vivo demonstration of this relationship, and they show that specific characteristics of aged skin such as the ratio of extracellular matrix components collagen and elastin can be evaluated by in vivo AF and SHG measurements using near-IR femtosecond laser pulses.