Airborne hyperspectral remote sensing (imaging spectroscopy) sensors acquire images of several (from tens to hundreds) narrow spectral bands in visible, near and short infrared wavelengths. So many narrow spectral bands allowed development of the advanced methods for Earth surface object detection with high accuracy identification of their physical and/or chemical properties. Use of hyperspectral remote sensing (RS) data in scientific and even commercial applications is quite broad, starting from agriculture, forestry, and natural vegetation (precision farming, assessment of general plant status, biomass estimation, species composition mapping), through geology (mapping of minerals, land degradation assessment), up to limnology (water quality evaluation), and other domains. Since 2004 the Institute of Systems Biology and Ecology (ISBE) (Academy of Sciences of the Czech Republic) has been operating the VNIR airborne hyperspectral sensor AISA Eagle. The AISA Eagle system, developed by Spectral Imaging (Specim Ltd, Finland) and mounted in an aircraft of the Argus Geo System Ltd., is a pushbroom imaging system with the spectral range from 400 to 1000 nm, the highest achievable spectral resolution of 2.9 nm, and spatial resolution between 0.4 m and 7.0 m. The spectral bands are during acquisition split by the slit prism and pixel-wise progressively recorded by the CCD matrix into the 12 bit digital numbers. The entire airborne imaging system consists of a compact hyperspectral sensor head, data acquisition unit (rugged PC), GPS/INS unit, and downwelling irradiance sensor (FODIS). This assembly, complemented by the supporting pre-processing software, is capable to deliver in almost real time the orhtorectified hyperspectral images in at surface and/or at sensor level reflectance values. Nevertheless, quality of the radiometric, geometric, and atmospheric corrections of the airborne hyperspectral images must always be validated. Therefore, the ground spectral and spatial reference data are measured simultaneously with the AISA Eagle over-flights using a portable field spectroradiometer ASD FieldSpec-3 and the geodetic Topcon GNSS system. The workgroup for remote sensing of vegetation at ISBE is currently capable to facilitate the complete flight/ground hyperspectral campaign including the standard image data pre-processing.