One of the most widely used methods to enable Homogeneous Charge Compression Ignition (HCCI) combustion is using negative valve overlapping to trap a sufficient quantity of hot residual gas. The characteristics of air motion with specially designed valve events having reduced valve lift and durations associated with HCCI engines and their effect on subsequent combustion are not yet fully understood. In addition, the ignition process and combustion development in such engines are very different from those in conventional spark-ignition or diesel compression ignition engines. Very little data has been reported concerning optical diagnostics of the flow and combustion in the engine using negative valve overlapping. This paper presents an experimental investigation into the in-cylinder flow characteristics and combustion development in an optical engine operating in HCCI combustion mode. PIV measurements have been taken under motored engine conditions to provide a quantitative flow characterisation of negative valve overlap in-cylinder flows. The ignition and combustion process was imaged using a high resolution charge coupled device (CCD) camera and the combustion imaging data was supplemented by simultaneously recorded in-cylinder pressure data which assisted the analysis of the images. It is found that the flow characteristics with negative valve overlapping are less stable and more valve event driven than typical spark ignition in-cylinder flows, while the combustion initiation locations are not uniformly distributed.