The use of spatial analysis in conservation biology as a research tool has grown tremendously over the past decade and a half. Although conservation biologists and ecologists have recognized the potential of spatial information for informing conservation biology and policy for a long time, such as for studying changes and trends in populations and habitats, it has been only recently that spatial analysis has been incorporated into most conservation biology studies. Since the 1990s, conservation biology has developed quickly by the application of spatial analysis technologies. In this paper, we review the history, methodologies and applications of this tool, and the potential for growth and other applications by using some projects and works in which the authors were involved as examples. First, we discussed the use of spatially explicit data on biodiversity and its distribution, and the significance of using spatially explicit methods in conservation biology was summarized. We presented patterns of biodiversity at the global scale and country level, and discussed plant diversity centers and vascular plant family diversity as monitored by the World Conservation Monitor Center (WCMC). We also discussed the spatial distribution of four groups (plant, birds, fishes and molluscs) of endangered species in the United States. Mapping the spatial distribution of biodiversity is a useful comparative tool for analyzing the patterns, magnitude and extent of biodiversity, changes in spatial distributions at different temporal scales, understanding the relationships between populations and habitats, and for conservation by spatial overlap analysis as in GAP analysis. Second, we reviewed various conservation projects including Global Forest Watch of World Resource Institute, National GAP Analysis of United States, Roadless Area of Forest Service-USA, and Nature Audit of Canada. Also, some examples from the literature were used, such as a comparative study of plant diversity richness between East Asia and North America and the spatial analysis of biological invasions. The spatial analysis of patterns of biodiversity and habitats were discussed in the third part of this paper. During the last two decades, pattern-oriented ecology and conservation biology has made a lot progress, especially spatial pattern analyses, spatial statistics originating from geo-statistics, geographic information systems, spatially explicit model-based growth of individuals (grid), population theory based on patch analysis (e.g., metapopulations and source-sink models), and so on. The application of spatial pattern analysis in conservation biology was summarized by examining two conservation projects: the forest fragmentation analysis of the USA and late seral forests spatial pattern analysis in the Pacific Northwest, USA. We also presented the theory of “Matrix conservation” by Lindenmayer and Franklin, “Conserving Forest Biodiversity, A Comprehensive Multiscaled Approach”(2002). We agree with the authors of this new initiative that extends conservation efforts beyond nature reserves to integrated conservation strategies that balance conservation and development at landscape or regional scales. Lastly, models that are used widely in conservation biology, the spatially explicit model, process-based spatial model, agent-based spatial adaptation model (SWAM) and Dynamics Global Vegetation Model (DGVM), were discussed.This new branch of conservation, spatial conservation biology, has matured as a new discipline that contains a lot of spatial and information technology and may make more contributions to the global biodiversity conservation.