The spider fauna at eight different types of forests of the Departamento del Valle del Cauca (southwestem Colombia) was sampled using diverse capture techniques. A total of 4500 specimens, grouped in 1498 morphospecies, 41 families and two suborders were collected. The most diverse families were Araneidae, Theridiidae, Salticidae, Tetragnathidae and Linyphiidae. Six families and 21 genera are recorded for the first time in Colombia. The microhabitats preferred by the sampled araneofauna are located in the low and shrubby vegetation. Hand collection, net sweeping and arboreal foliage beating were the most effective techniques for sampling spider diversity.