The mechanisms of cold injury to biosystems are investigated at the cellular, tissue, and systemic levels, with details concerning the related experimental methods. Cellular level studies have shown a direct relationship between biophysical changes versus freeze-thaw survival. Advances in experimental and analytical methods have resulted in qualitatively similar results for native and artificial tissue systems, with several important caveats relating to cell-cell, cell-ECM effects. While these biophysical events have informed a better understanding of immediate injury after freezing at the cell and tissue level, further understanding of delayed injury effects after cryosurgery including at the cellular (i.e. apoptosis) and host mediated (i.e. vascular and immunological) events remain important areas of research and an opportunity to improve the technique.