Boundary lubrication is one of the most interesting topics in the field of tribology, and many studies have been conducted in the past to gain deeper understanding of the behaviour of boundary lubrication layers formed by additives mixed in lubricants because the layer largely affects the tribological property of the target surface. With a strong demand to clarify the behaviour of boundary lubrication layers, several new approaches to in situ (=in lubricant) analysis have been proposed and applied. This paper focuses on the effectiveness of combined use of neutron reflectometry and frequency-modulation atomic force microscopy for the quantitative analysis of boundary lubrication layers formed at the interface between the metal substrate and lubricant and introduced their application with concrete case examples.