Hyperspectral remote sensing has great potential for accurate retrieval of forest biochemical parameters. In this paper, a hyperspectral remote sensing algorithm is developed to retrieve total leaf chlorophyll content for both open spruce and closed forests, and tested for open forest canopies. Ten black spruce ( Picea mariana (Mill.)) stands near Sudbury, Ontario, Canada, were selected as study sites, where extensive field and laboratory measurements were carried out to collect forest structural parameters, needle and forest background optical properties, and needle biophysical parameters and biochemical contents chlorophyll a and b. Airborne hyperspectral remote sensing imagery was acquired, within one week of ground measurements, by the Compact Airborne Spectrographic Imager (CASI) in a hyperspectral mode, with 72 bands and half bandwidth 4.25–4.36 nm in the visible and near-infrared region and a 2 m spatial resolution. The geometrical–optical model 4-Scale and the modified leaf optical model PROSPECT were combined to estimate leaf chlorophyll content from the CASI imagery. Forest canopy reflectance was first estimated with the measured leaf reflectance and transmittance spectra, forest background reflectance, CASI acquisition parameters, and a set of stand parameters as inputs to 4-Scale. The estimated canopy reflectance agrees well with the CASI measured reflectance in the chlorophyll absorption sensitive regions, with discrepancies of 0.06%–1.07% and 0.36%–1.63%, respectively, in the average reflectances of the red and red-edge region. A look-up-table approach was developed to provide the probabilities of viewing the sunlit foliage and background, and to determine a spectral multiple scattering factor as functions of leaf area index, view zenith angle, and solar zenith angle. With the look-up tables, the 4-Scale model was inverted to estimate leaf reflectance spectra from hyperspectral remote sensing imagery. Good agreements were obtained between the inverted and measured leaf reflectance spectra across the visible and near-infrared region, with R 2 = 0.89 to R 2 = 0.97 and discrepancies of 0.02%–3.63% and 0.24%–7.88% in the average red and red-edge reflectances, respectively. Leaf chlorophyll content was estimated from the retrieved leaf reflectance spectra using the modified PROSPECT inversion model, with R 2 = 0.47, RMSE = 4.34 μg/cm 2, and jackknifed RMSE of 5.69 μg/cm 2 for needle chlorophyll content ranging from 24.9 μg/cm 2 to 37.6 μg/cm 2. The estimates were also assessed at leaf and canopy scales using chlorophyll spectral indices TCARI/OSAVI and MTCI. An empirical relationship of simple ratio derived from the CASI imagery to the ground-measured leaf area index was developed ( R 2 = 0.88) to map leaf area index. Canopy chlorophyll content per unit ground surface area was then estimated, based on the spatial distributions of leaf chlorophyll content per unit leaf area and the leaf area index.