Water quality, size, connectivity and other physical properties of hydrological systems might have different functions in the formation and maintenance of biodiversity, but this remains mostly unclear due to the lack of undisturbed sites for experimental modelling. Alpine freshwater habitats such as micro-waterbodies (MWB) represent a kind of natural system suitable for biodiversity research. In order to assess potential linkages between environmental factors, connectivity of MWBs and aquatic species richness, we conducted a pilot study in two separated MWB systems located in Gaoligong Mountain, northwest Yunnan province, China. A total of 27 MWBs have been analyzed, including 22 connected and 5 isolated bodies. 13 conventional environmental factors were tested while all kinds of aquatic macro-organisms were collected and classified. Results showed a high environmental heterogeneity among MWBs and significant differences between the two systems but only a few environmental variables such as the depth of soil bottom, total Nitrogen and altitude were related to species richness and the formation of the community structure. As a benefit from the high environmental heterogeneity, the cascaded MWB systems provided divergent habitats able to support species richness at a higher level than the same number of randomly selected MWBs. This finding supports the idea that habitat connectivity matters also in extremely small aquatic ecosystems. Moreover, although still a preliminary result, a polarization effect within connected chains where edge MWBs host higher taxa and endemic taxa richness as well as larger populations, was detectable. This study gives interesting insights on the spatial processes driving community structure and a new prospective for biodiversity conservation. Since alpine MWBs have significant effects on the maintenance of watershed biodiversity, further research on such small and crucial ecosystems is encouraged.