Collections of tissue samples stand as keystone sources of molecular information to construct biodiversity knowledge, and are particularly useful in megadiverse countries. In 1998 the Humboldt Institute (<em>Instituto de Investigación de Recursos Biológicos Alexander von Humboldt</em>) began a tissue collection of Colombian biodiversity (IAvH-CT) and the aim of this work is to present a diagnostic and an historical narrative for that collection, constructed by compiling information and experiences on its management as well as by organizing and curating the information of each catalogued sample. After 16 years the IAvH-CT harbors 16469 samples, which represent around 2530 species from 1289 genera, and 323 families of the Colombian biodiversity. Samples are biased toward plants and birds (84 %), but also include other animal taxa. Geographically, IAvH-CT includes samples from all Colombian departments, but there is a broad variation in their coverage. When compared with other international collections IAvH-CT fulfills several standards of sample storage and data management, but have a major weakness that is that several tissues seem to lack a vouchered specimen. Tissues housed at IAvH-CT have been included in at least 48 studies published in several scientific journals. IAvH-CT is implementing strategies to improve curatorial standards, fill-up taxonomic gaps, and to explore the potential of its samples to understand the outstanding Colombian biota in a cooperative research frame among institutions.