A low-temperature atomic force microscope (cryo-AFM), operated in liquid nitrogen vapor, has been constructed for biological applications. The system provides an adjustable imaging temperature from 77 to 220 K with atomic resolution achieved on crystalline specimens. Imaging with NaCl microcrystals demonstrates that the system is free from surface contamination. Below 100 K, several biological specimens, including immunoglobulins and DNA as well as red blood cell ghosts, were imaged at high spatial resolution. Measurements on individual macromolecules showed that the mechanical strength is significantly greater at cryogenic temperatures with an estimated Young's modulus 1000-10,000 times that of a hydrated protein at room temperature, providing a solid basis for future improvements and applications of cryo-AFM in structural biology.