Devices and methods are provided that permit efficient and selective separation of liquid biological specimens into at least two constituent components to facilitate subsequent quantitative and qualitative analysis on at least one analyte of interest in at least one of the components. The devices generally include one or more sample deposition regions supported on a base. Each sample deposition region includes a separation membrane for separating the liquid biological specimen into two different fractions. The first fraction is trapped by the separation membrane while the second fraction passes through the separation membrane and into a respective collection membrane. The separation and collection membranes are easily separable from the devices and can be utilized for further processing and analysis.