Biochemicals, such as chlorophyll (Chl) and nitrogen (N), are closely related to photosynthesis process of vegetation. Their accurate estimation is an important topic in remote sensing of vegetation. Previous studies mainly focused on Chl-N content inversion in leaf and canopy level, and few cared about their 3D distributions, which was also an important indicator for the growth status of vegetation (GSV). Hyperspectral LiDAR (HSL) is a novel active remote sensing technology, which has target-sensitive band with hyper-spectra resolution. Its 3D point cloud data simultaneously contains rich spectral and precise geometrical characteristics of the target. This work aims to apply HSL data on 3D Chl-N content mapping in vegetation through constructing HSL-based spectral indices (SIs). Except for following the SI forms of previous works, the normalized differential vegetation index (NDVI) and ratio index (RI) with four-broad-band in an HSL spectral space were successively proposed to invert Chl-N content for the whole vegetation based on the artificial neural network (ANN) method. These four broad-bands were transformed based on the relative spectral response curve of detector and the feature weights (FWs) of multiwavelength respectively. Results show that most HSL-based ANN models can accurately invert Chl-N content with a mean R 2 of >0.75, and some that fusing broad-band data with convolution transformation, namely the FW-based RI, can even obtain a model R 2 of 0.84 for N content inversion. Thus, HSL can be efficiently applied to 3D Chl-N content mapping of vegetation and has great potential in GSV monitoring.